**7. References**

52 Steroids – Basic Science

RNA can be used with Illumina, Applied Biosystems SOLIiD and Roche 454 Life Science platforms (Wang et al., 2009, Rev). RNA-Seq has very low, if any, background signal because cDNA sequences can be mapped to unique regions of the genome. It does not have any upper limit of quantification like DNA microarrays which lack sensitivity for genes expressed either at low or very high levels. Like other HTP Sequencing technologies, RNA-

The analysis of RNA-Seq data starts from raw cDNA sequences, usually having lengths of 40-70bp, depending on the platform. The general goal is to find which sites in human genome the RNA was transcribed from and determine the expression levels of these transcripts. Additionally, RNA-Seq data can be used to study expression levels of alternative

1. Map each RNA sequence to the human genome. Mapping program has to enable spliced alignments because the sequences can come from separated exons. For this a

2. Count mappings to every site and measure the expression level of the sites. The expression level is usually measured in **F**ragments **P**er **K**ilobase of exon per **M**illion fragments mapped (FPKM) which permits comparison of results across experiments. FPKM of a site shows how frequently the mapped fragments fall on that site. This analysis is possible using Cufflinks software, which can measure FPKM for whole genes

3. Compare experiments and find genes (or spliced isoforms) that have statistically significant different FPKM across experiments. This is also possible using commercial

This chapter summarised the current knowledge of E2 and P4 action in human endometrium and mammary gland. To understand both sides of steroid hormone action - in normal physiology and especially in pathology it is important to understand the molecular intracellular events of E2 and P4 in tissue-type manner. The same hormones could have different or even opposite effects in different tissues. Thus, attention should be applied to steroid hormones' or their analogues' possible side effects before using them in clinical treatments. Questions still remain about the aberrations of the endometrium leading to implantation failure, endometriosis and other dys-regulations. Even though studies of breast cancer development continuously unravel new information about the mechanisms leading from normal to malignant tissue proliferation, breast cancer is still the most fatal cancer type among women. The number of couples seeking for aid to achieve fertility is constantly increasing and thus a better understanding of factors needed for successful

We would like to punctuate the importance of the next generation sequencing technologies which we believe are the key in understanding hormone dependent action in whole organism. With today´s knowledge of nearly entire human genome sequence and the development of new technologies based on HTP sequencing it has become possible to define all targets for the TF in vivo and establish entire transcriptome in a single experiment. Data

fast and open source tool TopHat can be used (Trapnell et al., 2009)

and also for specific spliced isoforms (Trapnell et al., 2010)

More detailed explanation of the analysis is depicted in the Figure 7.

seq faces several bioinformatics challenges in data processing.

The usual analysis consists of three main steps:

or open source software as Cufflinks.

treatment and possible side effects is crucial.

splicing isoforms.

**5. Conclusion** 


The Tissue Specific Role

95: 104–110.

Report 2001

84-90.

2006;50/4:720-734.

Aug;17(8):2209-13.

of Estrogen and Progesterone in Human Endometrium and Mammary Gland 55

Check, J.H., O'Shaughnessy, A., Lurie, D., Fisher, C. & Adelson, H.G. (1995). Evaluation of

Cohen, F. J., Watts, S., Shah, A., Akers, R., and Plouffe, L., Jr. (2000) *Uterine effects of 3-year* 

Cohen, I. (2004) *Endometrial pathologies associated with postmenopausal tamoxifen treatment.* 

Cork, B.A., Li, T.C., Warren, M.A. & Laird SM. (2001) *Interleukin-11 (IL-11) in human* 

*endometrial IL-11 production in vitro.* J Reprod Immunol. 2001 Apr;50(1):3-17. Dadi, T.D., Li, M.W. & Lloyd K.C. (2006) *Development of mouse embryos after* 

Dauvois, S., White, R. and Parker, M.G. (1993) *The antiestrogen ICI 182780 disrupts estrogen receptor nucleocytoplasmic shuttling.* Journal of Cell Science 1993; 106: 1377-1388. Dekker, J., Rippe, K., Dekker, M. & Kleckner N (2002) *Capturing chromosome conformation.*

Department of Health and Human Services Centres for Disease Control and Prevention

Develioglu, O.H., Hsiu, J.G., Nikas, G., Toner, J.P., Oehninger, S. and Jones, H.W. Jr (1999).

Devroey, P., Bourgain, C., Macklon, N.S. & Fauser, B.C. (2004). Reproductive biology and

Dietz, SC. & Carroll, JS. (2008) *Interrogating the genome to understand oestrogen-receptormediated transcription.* Expert Rev Mol Med. 2008 Apr 1;10:e10. Review. Diez-Perez, A. (2006*) Selective Estrogen Receptor Modulators.* Arq Bras Endocrinol Metab

Dimitriadis, E., Salamonsen, L.A. & Robb L. (2000) *Expression of interleukin-11 during the* 

Doyle, P., Maconochie, N., Beral, V., Swerdlow, A.J. & Tan SL. (2002*) Cancer incidence* 

Duan, L., Glass, CK., Rosenfeld MG. & Fu XD (2007) *Sensitive ChIP-DSL technology reveals an* 

Acad Sci U S A. 2007 Mar 20;104(12):4852-7. Epub 2007 Mar 14.

stimulated cycles of oocyte donors. *Fertil. Steril.*, 71, 1040-1047.

Endometrial estrogen and progesterone receptor and pinopode expression in

IVF: ovarian stimulation and endometrial receptivity. *Trends Endocrinol. Metab.,* 15,

*human menstrual cycle: coincidence with stromal cell decidualization and relationship to leukaemia inhibitory factor and prolactin.* Mol Hum Reprod. 2000 Oct;6(10):907-14. Dor, J., Lerner-Geva, L., Rabinovici, J., Chetrit, A., Levran, D., Lunenfeld, B., Mashiach, S. &

Modan B. (2002) *Cancer incidence in a cohort of infertile women who underwent in vitro* 

*following treatment for infertility at a clinic in the UK.* Hum Reprod. 2002

*extensive estrogen receptor alpha-binding program on human gene promoters.* Proc Natl

oocyte programme. *Hum. Reprod*., 10, 3022–3027.

Gynecology and Oncology 2004;94: 256–266.

Med. 2006 Jun;56(3):188-95.

Science. 2002 Feb 15;295(5558):1306-11.

*fertilization.* Fertil Steril. 2002 Feb;77(2):324-7.

the mechanism for higher pregnancy rates in donor oocyte recipients by comparison of fresh with frozen embryo transfer pregnancy rates in a shared

*raloxifene therapy in postmenopausal women younger than age 60.* Obstet. Gynecol. 2000;

*endometrium: expression throughout the menstrual cycle and the effects of cytokines on* 

*immunoneutralization of mitogenic growth factors mimics that of cloned embryos.* Comp


Berrodin, T.J., Chang, K.C., Komm, B.S., Freedman, L.P. & Nagpal S. (2009) *Differential* 

Berry, M., Metzger, D. and Chambon, P. (1990) *Role of the two activating domains of the* 

Björnström, L. & Sjöberg, M. (2004*) Estrogen receptor-dependent activation of AP-1 via non-*

Bocchinfuso, W.P. & Korach KS. (1997) *Mammary gland development and tumorigenesis in* 

Bock, C. & Lengauer T. (2008). *Computational epigenetics.* Jan 1;24(1):1-10. Epub 2007 Nov 17.

Bowler, J., Lilley, T.J., Pittam, J.D. and Wakeling, A.E. (1989) *Novel steroidal pure antiestrogens.*

Brinsden, P.R., Alam, V., de Moustier, B. & Engrand P. (2009) *Recombinant human leukemia* 

Brinton, L.A., Scoccia, B., Moghissi, K.S., Westhoff, C.L., Althuis, M.D., Mabie, J.E. & Lamb

Brogden, R.N., Goa, K.L. & Faulds D*.* (1993) *Mifepristone: a review of its pharmacodynamic and pharmacokinetic properties, and therapeutic potential.* Drugs. 1993;45:384-409. Brosens, J.J. & Gellersen, B. (2006) *Death or survival--progesterone-dependent cell fate decisions in the human endometrial stroma.* J Mol Endocrinol. 2006 Jun;36(3):389-98. Review. Buzdar, A. (1998) *The place of chemotherapy in the treatment of early breast cancer.* British Journal

Cakmak, H.& Taylor, H.S. (2010). Review. Molecular mechanisms of treatment resistance in

Carroll, JS. & Brown M. (2006) *Estrogen receptor target gene: an evolving concept.* Mol Cell Biol.

Carson, DD., Lagow, E., Thathiah, A., Al-Shami, R., Farach-Carson, MC., Vernon, M., Yuan,

Casneuf, T., Van de Peer, Y. & Huber W. *In situ analysis of cross-hybridisation on microarrays and the inference of expression correlation.* BMC Bioinformatics. 2007 Nov 26;8:461. Chang, Y., Lee, J.J., Chen, W.F., Chou, D.S., Huang, S.Y. & Sheu JR. (2010) *A novel role for* 

*microarray screening.* Mol Hum Reprod. 2002 Sep;8(9):871-9.

endometriosis: the role of progesterone-hox gene interactions. *Semin. Reprod. Med*.,

L., Fritz MA. & Lessey. BA. (2002) *Changes in gene expression during the early to midluteal (receptive phase) transition in human endometrium detected by high-density* 

*tamoxifen in the inhibition of human platelets.* Transl Res. 2011 Feb;157(2):81-91. Epub

*anti-oestrogen 4- hydroxytamoxifen.* EMBO J. 1990;9: 2811-2818.

*genomic signalling.* Nucl Recept. 2004 Jun 14;2(1):3.

Steril. 2009 Apr;91(4 Suppl):1445-7. Epub 2008 Aug 16.

2004 Sep;19(9):2005-13. Epub 2004 Jun 24.

2007 Jul;27(14):5090-104. Epub 2007 May 21.

Epub 2008 Nov 26.

34.

Bioinformatics.

Steroids 1989;54:71-99.

of Cancer 1998; 78

1 (January, 28), 69-74.

2010 Nov 30.

*biochemical and cellular actions of Premarin estrogens: distinct pharmacology of bazedoxifene-conjugated estrogens combination.* Mol Endocrinol. 2009 Jan;23(1):74-85.

*oestrogen receptor in the cell-type and promoter-context dependent agonistic activity of the* 

*estrogen receptor knockout mice.* J Mammary Gland Biol Neoplasia. 1997 Oct;2(4):323-

*inhibitory factor does not improve implantation and pregnancy outcomes after assisted reproductive techniques in women with recurrent unexplained implantation failure.* Fertil

EJ. (2004) *Breast cancer risk associated with ovulation-stimulating drugs.* Hum Reprod.


The Tissue Specific Role

2008 Oct 3.

of Estrogen and Progesterone in Human Endometrium and Mammary Gland 57

Haouzi, D., Assou, S., Mahmoud, K., Tondeur, S., Rème, T., Hedon, B., De Vos, J. &

Haouzi, D., Mahmoud, K., Fourar, M., Bendhaou, K., Dechaud, H., De Vos, J., Rème, T.,

Hofmann, G.E., Scott, R.T. Jr., Bergh, P.A. & Deligdisch L. (1991) *Immunohistochemical* 

Horcajadas, J.A., Mínguez, P., Dopazo, J., Esteban, F.J., Domínguez, F., Giudice, L.C., Pellicer, A. & Simon, C. (2008). *J. Clin. Endocrinol. Metab.,* 93, 4500–4510. Horcajadas, J.A., Mínguez, P., Dopazo, J., Esteban, F.J., Domínguez, F., Giudice, L.C.,

Horcajadas, J.A., Riesewijk, A., Martín, J., Cervero, A., Mosselman, S., Pellicer, A. & Simón

Humphreys, R.C., Lydon, J., O'Malley, B.W. & Rosen JM. (1997*) Mammary gland development* 

Johnston, S.J. & Cheung, K.L. (2010) *Fulvestrant—a novel endocrine therapy for breast cancer.*

Jones, R.L., Stoikos, C., Findlay, J.K. & Salamonsen, L.A. (2006) *TGF-beta superfamily* 

Jordan, C. (2001) *Selective Estrogen Receptor Modulator: A Personal Perspective.* Cancer Res

Jordan, VC., Gapstur, S. & Morrow M. (2001) *Selective Estrogen Receptor Modulation and* 

Journal of the National Cancer Institute 2001; Vol. 93, No. 19: 1449-57. Kabir-Salmani, M., Shiokawa, S., Akimoto, Y., Sakai, K. & Iwashita M. (2004) *The role of* 

*during the process of implantation.* Mol Hum Reprod. 2004 Feb;10(2):91-7. Källén, B., Finnström, O., Lindam, A., Nilsson, E., Nygren, K.G. & Olausson PO. (2011)

Källén, B., Finnström, O., Nygren, K.G. & Olausson PO. (2007) *In vitro fertilization (IVF) in* 

Kanis, JA. & Stevenson, JC. (1994). Effect of estrogen therapy on bone density in elderly

women. N Engl J Med 1994 Mar 10;330(10):715; author reply 716

2009 Jun;24(6):1436-45. Epub 2009 Feb 26.

Clin Endocrinol Metab. 1991 Oct;73(4):882-7.

Metab. 2008 Nov;93(11):4500-10. Epub 2008 Aug 12

Immunol. 2004 Aug;63(1):41-9.

Curr Med Chem 2010, 17:902–914.

Jan;26(1):253-8. Epub 2010 Nov 18.

A Clin Mol Teratol. 2005 Mar;73(3):162-9.

Aug;132(2):217-32. Review.

2001; 61: 5683–5687

Jun;11(6):801-11.

Hamamah S. (2009b) *Gene expression profile of human endometrial receptivity: comparison between natural and stimulated cycles for the same patients.* Hum Reprod.

Dewailly, D. & Hamamah S. (2009) *Identification of new biomarkers of human endometrial receptivity in the natural cycle.* Hum Reprod. 2009 Jan;24(1):198-205. Epub

*localization of epidermal growth factor in human endometrium, decidua, and placenta.* J

Pellicer, A. & Simón C. (2008) *Controlled ovarian stimulation induces a functional genomic delay of the endometrium with potential clinical implications.* J Clin Endocrinol

C. (2004) *Global gene expression profiling of human endometrial receptivity.* J Reprod

*is mediated by both stromal and epithelial progesterone receptors.* Mol Endocrinol. 1997

*expression and actions in the endometrium and placenta.* Reproduction. 2006

*Reduction in Risk of Breast Cancer, Osteoporosis, and Coronary Heart Disease.* Review.

*alpha(5)beta(1)-integrin in the IGF-I-induced migration of extravillous trophoblast cells* 

*Malignancies among women who gave birth after in vitro fertilization.* Hum Reprod. 2011

*Sweden: risk for congenital malformations after different IVF methods.* Birth Defects Res


Edwards, R.G. (2005) Historical significance of gonadotrophins in assisted reproduction.

Enmark. E., Pelto-Huikko, M., Grandien K., Lagercrantz, S., Lagercrantz, J., Fried, G.,

Fauser, B.C. & Devroey, P. (2003). Reproductive biology and IVF: ovarian stimulation and

Fisher, B., Costantino, JP., Wickerham, DL., Redmond, CK., Kavanah, M., Cronin, WM.,

Fisher, B., Costantino, JP., Wickerham, DL., Redmond, CK., Kavanah, M., Cronin, WM.,

Frasor, J., Danes, JM., Komm, B., Chang, KC., Lyttl,e CR. & Katzenellenbogen BS. (2003)

Fullwood, MJ., Liu, MH., Pan, YF., Liu, J., Xu, H., Mohamed, YB., Orlov, YL., Velkov, S., Ho,

*bound human chromatin interactome.* Nature. 2009 Nov 5;462(7269):58-64. Gibson, M.K., Nemmers, L.A., Beckman, W.C. Jr., Davis, V.L., Curtis, S.W. and Korach, K.S.

Giudice, L.C., Telles, T.L., Lobo, S. & Kao, L. (2002). The molecular basis for implantation

Goldberg, J.R., Plescia, M.G. & Anastasio, G.D. (1998) *Mifepristone (RU 486). Current* 

Graham, R.A., Seif, M.W., Aplin, J.D., Li, T.C., Cooke, I.D., Rogers, A.W. & Dockery P. (1990) *An endometrial factor in unexplained infertility.* BMJ. 1990 Jun 2;300(6737):1428-31. Hall, JM. & McDonnell DP (2005). *Coregulators in nuclear estrogen receptor action: from concept* 

*receptor in uterine tissue.* Endocrinology 1991;129:2000-2010.

*Knowledge and Future Prospects.* Arch Fam Med. 1998;7:219-222.

*to therapeutic targeting.* Mol Interv. 2005 Dec;5(6):343-57. Review.

*in postmenopausal women*. Am. J. Obstet. Gynecol. 2000;182: 568–574.

luteal phase consequences. *Trends. Endocrinol. Metab.*, 14, 236-242.

Nordenskjöld, M. & Gustafsson, JA. (1997) *Human estrogen receptor beta-gene structure, chromosomal localization, and expression pattern.* J Clin Endocrinol Metab.

Vogel, V., Robidoux, A., Dimitrov, N., Atkins, J., Daly, M., Wieand, S., Tan-Chiu, E., Ford, L. & Wolmark N. (1998) *Tamoxifen for prevention of breast cancer: report of the National Surgical Adjuvant Breast and Bowel Project P-1 Study.* J Natl Cancer Inst.

Vogel, V., Robidoux, A., Dimitrov, N., Atkins, J., Daly, M., Wieand, S., Tan-Chiu, E., Ford, L. & Wolmark, N. (2005) *Tamoxifen for the Prevention of Breast Cancer: Current Status of the National Surgical Adjuvant Breast and Bowel Project P-1 Study.* J Natl

*Profiling of estrogen up- and down-regulated gene expression in human breast cancer cells: insights into gene networks and pathways underlying estrogenic control of proliferation and cell phenotype.* Endocrinology. 2003 Oct;144(10):4562-74. Epub 2003 Jul 10. Fugere, P., Scheele, W. H., Shah, A., Strack, T. R., Glant, M. D., and Jolly, E. (2000) *Uterine* 

*effects of raloxifene in comparison with continuous-combined hormone replacement therapy* 

A., Mei, PH., Chew, EG., Huang, PY., Welboren, WJ., Han, Y., Ooi, HS., Ariyaratne, PN., Vega, VB., Luo, Y., Tan, PY., Choy, PY., Wansa, KD., Zhao, B., Lim, KS., Leow, SC., Yow, JS., Joseph, R., Li, H., Desai, KV., Thomsen, JS., Lee, YK., Karuturi, RK., Herve, T., Bourque, G., Stunnenberg, HG., Ruan, X., Cacheux-Rataboul, V., Sung, WK., Liu, ET., Wei, CL., Cheung, E. & Ruan, Y. (2009) *An oestrogen-receptor-alpha-*

(1991) *The mechanism of ICI 164,384 antiestrogenicity involves rapid loss of estrogen* 

failure in endometriosis: on the road to discovery. *Ann. N Y Acad. Sci.,* 955, 252–

*Reprod. BioMed Online*, 10, 3

1997 Dec;82(12):4258-65.

1998; 90 (18): 1371-1388.

264.

Cancer Inst 2005;97(22):1652-1662.


The Tissue Specific Role

27.

Oct 28.

23;410(1):87-90. Review.

Ye, Z., Bibikova, M., Fan, JB.,

*Reprod*., 13, 3203-3205.

Nov;6(6):831-9. Review.

May;61(5):812-4.

Aug;62(4):446-55.

Mar 6;9:29.

Review.

of Estrogen and Progesterone in Human Endometrium and Mammary Gland 59

Kristiansson, P., Björ, O. & Wramsby H. (2007) *Tumour incidence in Swedish women who gave* 

Kuiper, GG. & Gustafsson, JA. (1997) *The novel estrogen receptor-beta subtype: potential role in* 

Kuokkanen, S., Chen, B., Ojalvo, L., Benard, L., Santoro, N. & Pollard JW. (2010) *Genomic* 

Kwon, YS., Garcia-Bassets, I., Hutt, KR., Cheng, CS., Jin, M., Liu, D., Benner, C., Wang, D.,

Lass, A., Peat, D., Avery, S. & Brinsden, P. (1998). Histological evaluation of endometrium

Lerner-Geva, L., Rabinovici, J. & Lunenfeld B. (2010) *Ovarian stimulation: is there a long-term* 

Lessey, B.A. (1994) *The use of integrins for the assessment of uterine receptivity.* Fertil Steril. 1994

Lessey, B.A. (2003) *Two pathways of progesterone action in the human endometrium: implications for implantation and contraception.* Steroids. 2003 Nov;68(10-13):809-15. Review Lessey, B.A. (2010) *Fine tuning of endometrial function by estrogen and progesterone through* 

Lessey, B.A., Gui, Y., Apparao, K.B., Young, S.L. & Mulholland J. (2002) *Regulated* 

Levenson, AS., Svoboda, KM., Pease, KM., Kaiser, SA., Chen, B., Simons, LA., Jovanovic,

*expressing wild-type estrogen receptor.* Cancer Res. 2002 Aug 1;62(15):4419-26. Li, R., Qiao, J., Wang, L., Li, L., Zhen, X., Liu, P. & Zheng X. (2011) *MicroRNA array and* 

Lim, H., Song, H., Paria, B.C., Reese, J., Das, S.K. & Dey SK. (2002). *Molecules in blastocyst* 

Lin, Z., Reierstad, S., Huang, CC. & Bulun SE. (2007) *Novel estrogen receptor-alpha binding* 

*breast cancer.* Cancer Res. 2007 May 15;67(10):5017-24.

*expression of heparin-binding EGF-like growth factor (HB-EGF) in the human endometrium: a potential paracrine role during implantation* Mol Reprod Dev. 2002

BD., Dyck. PA. & Jordan VC. (2002) *Gene expression profiles with activation of the estrogen receptor alpha-selective estrogen receptor modulator complex in breast cancer cells* 

*microarray evaluation of endometrial receptivity in patients with high serum progesterone levels on the day of hCG administration.* Reprod Biol Endocrinol. 2011

*implantation: uterine and embryonic perspectives.* Vitam Horm. 2002;64:43-76.

*sites and estradiol target genes identified by chromatin immunoprecipitation cloning in* 

*microRNAs.* Biol Reprod. 2010 Apr;82(4):653-5. Epub 2010 Feb 3.

*birth following IVF treatment.* Hum Reprod. 2007 Feb;22(2):421-6. Epub 2006 Oct

*the cell- and promoter-specific actions of estrogens and anti-estrogens.* FEBS Lett. 1997 Jun

*profiling of microRNAs and messenger RNAs reveals hormonal regulation in microRNA expression in human endometrium.* Biol Reprod. 2010 Apr;82(4):791-801. Epub 2009

on the day of oocyte retrieval after gonadotrophin-releasing hormone agonistfollicle stimulating hormone ovulation induction for invitro fertilization. *Hum.* 

*risk for ovarian, breast and endometrial cancer?* Womens Health (Lond Engl). 2010


Kao, L.C., Germeyer, A., Tulac, S., Lobo, S., Yang, J.P., Taylor, R.N., Osteen, K., Lessey, B.A.

Kao, L.C., Tulac, S., Lobo, S., Imani, B., Yang, J.P., Germeyer, A., Osteen, K., Taylor, R.N.,

Kao, LC., Tulac, S., Lobo, S., Imani, B., Yang, JP., Germeyer, A., Osteen, K., Taylor, RN.,

Kastner, P., Krust, A., Turcotte, B., Stropp, U., Tora, L. Gronemeyer, M.A. & Chambon, P.

Kastner. P., Bocquel. MT., Turcotte, B., Garnier, JM., Horwitz, KB., Chambon P. &

Kelly, MJ. & Rønnekleiv OK. (2008). *Membrane-initiated estrogen signaling in hypothalamic* 

Khan, S.A., Rogers, M.A., Obando, J.A. & Tamsen A. (1994) *Estrogen receptor expression of* 

Kim, J.J., Taylor, H.S., Lu, Z., Ladhani, O., Hastings, J.M., Jackson, K.S., Wu, Y., Guo, S.W. &

Kininis, M., Chen, BS., Diehl, AG., Isaacs, GD., Zhang, T., Siepel, AC., Clark, AG. & Kraus

Kotsopoulos, J., Librach, C.L., Lubinski, J., Gronwald, J., Kim-Sing, C., Ghadirian, P., Lynch,

Krikun, G., Schatz, F., Taylor, R., Critchley, H.O., Rogers, P.A., Huang, J. & Lockwood CJ.

role in decidualization. *Mol. Hum. Reprod*., 5 (May, 13), 323-32.

Endocrinol. 2006 Aug;20(8):1707-14. Epub 2006 Jan 5. Review.

Control. 2008 Dec;19(10):1111-9. Epub 2008 May 29.

*generating two receptor isoforms.* J Biol Chem. 1990 Jul 25;265(21):12163-7. Katz, D., Paltiel, O., Peretz, T., Revel, A., Sharon, N., Maly, B., Michan, N., Sklair-Levy, M. &

*the window of implantation.* Endocrinology. 2002 Jun;143(6):2119-38.

*the window of implantation.* Endocrinology. 2002 Jun;143(6):2119-38.

infertility. *Endocrinol.,* 144, 2870–2881

*9(5), 1603–1614.*

Oct 14.

Review.

15;54(4):993-7.

& Giudice, L.C. (2003). Expression profiling of endometrium from women with endometriosis reveals candidate genes for disease-based implantation failure and

Lessey, B.A. & Giudice LC. (2002). *Global gene profiling in human endometrium during* 

Lessey, BA. & Giudice LC. (2002) *Global gene profiling in human endometrium during* 

(1990) Two distinct estrogen-regulated promoters generate transcripts encoding the two functionally different human progesterone receptor forms A and B. *Embo*. *J.,* 

Gronemeyer H. (1990) *Transient expression of human and chicken progesterone receptors does not support alternative translational initiation from a single mRNA as the mechanism* 

Allweis T. (2008) *Beginning IVF treatments after age 30 increases the risk of breast cancer: results of a case-control study.* Breast J. 2008 Nov-Dec;14(6):517-22. Epub 2008

*neurons.* Mol Cell Endocrinol. 2008 Aug 13;290(1-2):14-23. Epub 2008 Apr 30.

*benign breast epithelium and its association with breast cancer.* Cancer Res. 1994 Feb

Fazleabas AT. (2007). Altered expression of HOXA10 in endometriosis: potential

WL. (2006) *Genomic analyses of transcription factor binding, histone acetylation, and gene expression reveal mechanistically distinct classes of estrogen-regulated promoters.* Mol

H.T., Moller, P., Foulkes, W.D., Randall, S., Manoukian, S., Pasini, B., Tung, N., Ainsworth, P.J., Cummings, S., Sun, P. &Narod, S.A. Hereditary Breast Cancer Clinical Study Group. *Infertility, treatment of infertility, and the risk of breast cancer among women with BRCA1 and BRCA2 mutations: a case-control study.* Cancer Causes

(2005) *Endometrial endothelial cell steroid receptor expression and steroid effects on gene expression.* J Clin Endocrinol Metab. 2005 Mar;90(3):1812-8. Epub 2004 Dec 21.


The Tissue Specific Role

of Estrogen and Progesterone in Human Endometrium and Mammary Gland 61

Park, PJ. (2009) *ChIP-seq: advantages and challenges of a maturing technology.* Nat Rev Genet.

Pennings G & Ombelet W. (2007). Coming soon to your clinic: patient-friendly ART. *Hum.* 

Potashnik, G., Lerner-Geva, L., Genkin, L., Chetrit, A., Lunenfeld, E. & Porath A. (1999)

Punyadeera, C., Dassen, H., Klomp, J., Dunselman, G., Kamps, R., Dijcks, F., Ederveen, A.,

Reese, J.C. and Katzenellenbogen, B.S. (1992*) Examination of the DNA-binding ability of* 

Riesewijk, A., Martín, J., van Os, R,, Horcajadas, JA., Polman, J., Pellicer, A., Mosselman,

Riggs, B.L. & Hartmann, L.C. (2003) *Selective estrogen-receptor modulators -- mechanisms of* 

Robertson, S.A., Mau, V.J., Young, I.G. & Matthaei KI. (2000) *Uterine eosinophils and* 

Roger, P., Sahla, M.E., Mäkelä, S., Gustafsson, J.A., Baldet, P. & Rochefort H. (2001) *Decreased* 

Russo, J., Calaf, G., Roi, L. & Russo IH. (1987) *Iinfluence of age and gland topography on cell kinetics of normal human breast tissue.* J Natl Cancer Inst. 1987 Mar;78(3):413-8. Seifert-Klauss, V. & Prior, JC (2010). *Progesterone and bone: actions promoting bone health in* 

Simon, C. (2008) *New perspectives in medically assisted procreation.* Gynecol Endocrinol. 2008

Simon, C., Cano, F., Valbuena, D., Remohi, J. & Pellicer, A. (1995). Clinical evidence for a

Simon, C., Oberyé, J., Bellver, J., Vidal, C., Bosch, E., Horcajadas, J.A., Murphy, C., Adams,

Smitz, J., Devroey, P., Faguer, B., Bourgain, C., Camus, M. & Van Steirteghem, A.C. (*1992*). A

high and normal responder patients. *Hum Reprod*, 10, 2432-2437.

detrimental effect on uterine receptivity of high serum oestradiol concentrations in

S., Riesewijk, A., Mannaerts, B. & Pellicer A. *Similar endometrial development in oocyte donors treated with either high- or standard-dose GnRH antagonist compared to treatment with a GnRH agonist or in natural cycles.* Hum Reprod. 2005 Dec;20(12):3318-27. Epub

prospective randomized comparison of intramuscular or intravaginal natural

*Fertility drugs and the risk of breast and ovarian cancers: results of a long-term follow-up* 

de Goeij, A. & Groothuis P. (2005) *Oestrogen-modulated gene expression in the human* 

*estrogen receptor in whole cells: implications for hormone-dependent transactivation and* 

S. & Simón C. (2003) *Gene expression profiling of human endometrial receptivity on days LH+2 versus LH+7 by microarray technology.* Mol Hum Reprod. 2003

*action and application to clinical practice.* N Engl J Med. 2003 Feb 13;348(7):618-29. Review. No abstract available. Erratum in: N Engl J Med. 2003 Mar

*reproductive performance in interleukin 5-deficient mice.* J Reprod Fertil. 2000

*expression of estrogen receptor beta protein in proliferative preinvasive mammary tumors.*

2009 Oct;10(10):669-80. Epub 2009 Sep 8.

*study.* Fertil Steril. 1999 May;71(5):853-9.

*endometrium.* Cell Mol Life Sci. 2005 Jan;62(2):239-50.

*the actions of antiestrogens.* Mol. Cell. Biol. 1992;12:4531-4538.

*Reprod.*, 22, 2075–2059.

May;9(5):253-64.

20;348(12):1192.

Nov;120(2):423-32.

Sep;24(9):485.

2005 Aug 5.

Cancer Res. 2001 Mar 15;61(6):2537-41.

*women.* J Osteoporos. 2010 Oct 31;2010:845180.


Lin, CY., Ström, A., Vega, VB., Kong, SL., Yeo, AL., Thomsen, JS., Chan, WC., Doray, B.,

Lin, CY., Vega, VB., Thomsen, JS., Zhang, T., Kong, SL., Xie, M., Chiu, KP., Lipovich, L.,

Liu, Y., Gao, H., Marstrand, TT., Ström, A., Valen, E., Sandelin, A., Gustafsson, JA. &

Matsuzaki, S., Murakami, T., Uehara, S., Canis, M., Sasano, H. & Okamura, K. (2001).

Mirkin, S., Arslan, M., Churikov, D., Corica, A., Diaz, JI., Williams, S., Bocca, S. & Oehninger

Mortazavi, A., Williams, BA., McCue, K., Schaeffer, L. & Wold B. (2008) Mapping and

Mote, P.A., Bartow, S., Tran, N. & Clarke CL. (2002) *Loss of co-ordinate expression of* 

Nargund, G. & Frydman, R. (2007). Towards a more physiological approach to IVF. *Reprod.* 

Nikas, G., Develioglu, O.H., Toner, J.P. and Jones, H.W. Jr (1999). Endometrial pinopodes

Olivennes, F., Fanchin, R., Ledee, N., Righini, C., Kadoch I.J. & Frydman, R. (2002). Perinatal

Papanikolaou, E.G., Bourgain, C., Kolibianakis, E., Tournaye, H., Devroey, P. (2005). Steroid

Pappo, I., Lerner-Geva, L., Halevy, A., Olmer, L., Friedler, S., Raziel, A., Schachter, M. &

the absence of secretory changes. *Hum. Reprod.*, 20, 1541–1547.

Surg Oncol. 2008 Apr;15(4):1048-55. Epub 2008 Jan 23.

*binding sites.* PLoS Genet. 2007 Jun;3(6):e87. Epub 2007 Apr 17.

Jul;6(7):33-8; discussion 38-40, 43. Review.

endometriosis. *Fertil.Steril.,* 75, 1198–1205.

Jul;5(7):621-8. Epub 2008 May 30.

Res Treat. 2002 Mar;72(2):163-72.

*Biomed Online*, 14, 550–552.

*Update*, 8, 117–128.

Epub 2004 Aug 12.

May 5.

792.

Bangarusamy, DK., Ramasamy, A., Vergara, LA., Tang, S., Chong, A., Bajic, VB., Miller, LD., Gustafsson, JA. & Liu ET. (2004) *Discovery of estrogen receptor alpha target genes and response elements in breast tumor cells.* Genome Biol. 2004;5(9):R66.

Barnett, DH., Stossi, F., Yeo, A., George, J., Kuznetsov, VA., Lee, YK., Charn, TH., Palanisamy, N., Miller, LD., Cheung, E., Katzenellenbogen, BS., Ruan, Y., Bourque, G., Wei, CL. & Liu ET. (2007) *Whole-genome cartography of estrogen receptor alpha* 

Dahlman-Wright K. *The genome landscape of ERalpha- and ERbeta-binding DNA regions.* Proc Natl Acad Sci U S A. 2008 Feb 19;105(7):2604-9. Epub 2008 Feb 13. Love, R.R. (1992) *Tamoxifen prophylaxis in breast cancer.* Oncology (Williston Park). 1992

Expression of estrogen receptor alpha and beta in peritoneal and ovarian

S. (2005) *In search of candidate genes critically expressed in the human endometrium during the window of implantation.* Hum Reprod. 2005 Aug;20(8):2104-17. Epub 2005

quantifying mammalian transcriptomes by RNA-Seq. Nat Methods. 2008

*progesterone receptors A and B is an early event in breast carcinogenesis.* Breast Cancer

indicate a shift in the window of receptivity in IVF cycles. *Hum. Reprod*., 14, 787-

outcome and developmental studies on children born after IVF. *Hum. Reprod.* 

receptor expression in late follicular phase endometrium in GnRH antagonist IVF cycles is already altered, indicating initiation of early luteal phase transformation in

Ron-El, R. (2008) *The possible association between IVF and breast cancer incidence.* Ann


The Tissue Specific Role

5.

Oct 1.

47.

14;346(8981):995-1000.

Nov;7(11):1378.

of Estrogen and Progesterone in Human Endometrium and Mammary Gland 63

Tung. L., Mohamed, MK., Hoeffler, JP., Takimoto, GS. & Horwitz, KB. (1993) *Antagonist-*

Ubaldi F, Rienzi L, Baroni E, Ferrero S, Iacobelli M, Minasi MG, Sapienza F, Romano S,

Valbuena, D., Martin, J., de Pablo, J.L., Remohi, J., Pellicer, A. & Simon, C. (2001). Increasing

Vandermolen, D.T. & Gu Y. (1996) *Human endometrial interleukin-6 (IL-6): in vivo messenger* 

Vegeto, E., Shahbaz, MM., Wen, DX., Goldman, ME., O'Malley, BW. & McDonnell DP.

Venn, A., Jones, P., Quinn, M. & Healy D. (2001) *Characteristics of ovarian and uterine cancers in a cohort of in vitro fertilization patients.* Gynecol Oncol. 2001 Jul;82(1):64-8. Venn, A., Watson, L., Lumley, J., Giles, G., King, C. & Healy D. (1995) *Breast and ovarian* 

von Rango, U., Alfer, J., Kertschanska, S., Kemp, B., Müller-Newen, G., Heinrich, P.C., Beier,

Wakeling, AE., Dukes, M. and Bowler, J. (1991) *A potent specific pure antiestrogen with clinical* 

Wang, Z., Gerstein, M. & Snyder M. (2009) *RNA-Seq: a revolutionary tool for transcriptomics.*

Weinmann, AS. & Farnham PJ. (2002). *Identification of unknown target genes of human* 

Wen, DX., , YF., Mais, DE., Goldman, ME. & McDonnell, DP. (1994) *The A and B isoforms of* 

Wolf, IM., Heitzer, MD., Grubisha, M. & DeFranco DB (2008*). Coactivators and nuclear receptor transactivation.* J Cell Biochem. 2008 Aug 1;104(5):1580-6. Review.

Weiss, BD. (1993*) RU 486: the progesterone antagonist.* Arch Fam Med. 1993;2:63-70.

*transcription factors using chromatin immunoprecipitation.* Methods. 2002 Jan;26(1):37-

*the human progesterone receptor operate through distinct signaling pathways within target* 

Biotechnol. 2010 May;28(5):511-5. Epub 2010 May 2.

stimulation. *Reprod Biomed Online*, 14, 675–681.

affect the embryo. *Fertil. Steril.,* 2001;76:962-968.

*beta.* Fertil Steril. 1996 Nov;66(5):741-7.

*potential.* Cancer Res. 1991;51:3867-3873.

Nat Rev Genet. 2009 Jan;10(1):57-63. Review.

*cells.* Mol Cell Biol. 1994 Dec;14(12):8356-64.

*reveals unannotated transcripts and isoform switching during cell differentiation.* Nat

*occupied human progesterone B-receptors activate transcription without binding to progesterone response elements and are dominantly inhibited by A-receptors.* Mol Endocrinol. 1993 Oct;7(10):1256-65. Erratum in: Mol Endocrinol 1993

Colasante A, Litwicka K. & Greco, E. (2007). Hopes and facts about mild ovarian

levels of estradiol are deleterious to embryonic implantation because they directly

*ribonucleic acid expression, in vitro protein production, and stimulation thereof by IL-1* 

(1993) *Human progesterone receptor A form is a cell- and promoter-specific repressor of human progesterone receptor B function.* Mol Endocrinol. 1993 Oct;7(10):1244-55. Venken, K., Callewaert F., Boonen, S. & Vanderschueren D (2008). *Sex hormones, their* 

*receptors and bone health.* Osteoporos Int. 2008 Nov;19(11):1517-25. Epub 2008 Apr

*cancer incidence after infertility and in vitro fertilisation.* Lancet. 1995 Oct

H.M. & Classen-Linke I (2004) *Interleukin-11 expression: its significance in eutopic and ectopic human implantation.* Mol Hum Reprod. 2004 Nov;10(11):783-92. Epub 2004

progesterone as a luteal phase and early pregnany supplementation. *Hum. Reprod.,*  7*,* 168*–175.*


Speirs, V., Carder, P.J. & Lansdown MR. (2002) *Oestrogen receptor beta: how should we measure* 

Spitz, I.M. & Bardin, C.W. (1993) *Mifepristone (RU 486): a modulator of progestin and* 

Stavreus-Evers, A., Aghajanova, L., Brismar, H., Eriksson, H., Landgren, B.M. & Hovatta O.

Stavreus-Evers, A., Nikas, G., Sahlin, L., Eriksson, H. & Landgren BM. (2001) *Formation of* 

Steck, T., Giess, R., Suetterlin, M.W., Bolland, M., Wiest, S., Poehls, U.G., & Dietl J. (2004)

Steptoe, P.C. & Edwards, R.G. (1978). *Birth After the Reimplantation of a Human Embryo*. The

Stewart, C.L. (1994) *Leukaemia inhibitory factor and the regulation of pre-implantation* 

Strathy, J.H., Molgaard, C.A., Coulam, C.B. & Melton, L.J. (1982). Endometriosis and

Tabibzadeh, S., Kong, Q.F., Babaknia, A. & May LT. (1995) *Progressive rise in the expression of* 

Talbi, S., Hamilton, A.E., Vo. KC., Tulac, S., Overgaard, M.T., Dosiou, C., Le Shay, N.,

Tamm, K., Rõõm, M., Salumets, A. & Metsis M (2009) *Genes targeted by the estrogen and* 

Taylor, H.S., Bagot, C., Kardana, A., Olive, D. & Arici, A. (1999). HOX gene expression is

Trapnell, C., Pachter, L. & Salzberg SL (2009) *TopHat: discovering splice junctions with RNA-*

Trapnell, C., Williams, BA., Pertea, G., Mortazavi, A., Kwan, G., van Baren, MJ., Salzberg,

*Seq.* Bioinformatics. 2009 May 1;25(9):1105-11. Epub 2009 Mar 16.

*implantation window.* Hum Reprod. 1995 Oct;10(10):2793-9.

*this?* Br J Cancer. 2002 Sep 9;87(6):687; author reply 688-9.

*glucocorticoid action.* N Engl J Med. 1993;329:404-412.

*progesterone receptors.* Fertil Steril. 2001 Oct;76(4):782-91.

Reprod Biol. 2004 Jan 15;112(1):69-73.

women. *Fertil. Steril., 6* (Dec, 38), 667-72.

Lancet, 312 (8085), 366.

121. Epub 2005 Nov 23.

Biol Endocrinol. 2009 Dec 24;7:150.

Review.

1331.

7*,* 168*–175.*

Aug;8(8):765-9.

progesterone as a luteal phase and early pregnany supplementation. *Hum. Reprod.,* 

(2002) *Co-existence of heparin-binding epidermal growth factor-like growth factor and pinopodes in human endometrium at the time of implantation.* Mol Hum Reprod. 2002

*pinopodes in human endometrium is associated with the concentrations of progesterone and* 

*Leukaemia inhibitory factor (LIF) gene mutations in women with unexplained infertility and recurrent failure of implantation after IVF and embryo transfer.* Eur J Obstet Gynecol

*development of the mammalian embryo.* Mol Reprod Dev. 1994 Oct;39(2):233-8.

infertility: a laparoscopic study of endometriosis among fertile and infertile

*interleukin-6 in human endometrium during menstrual cycle is initiated during the* 

Nezhat, C.N., Kempson, R., Lessey, B.A., Nayak, N.R. & Giudice LC. *Molecular phenotyping of human endometrium distinguishes menstrual cycle phases and underlying biological processes in normo-ovulatory women.* Endocrinology. 2006 Mar;147(3):1097-

*progesterone receptors in the human endometrial cell lines HEC1A and RL95-2.* Reprod

altered in the endometrium of women with endometriosis. *Hum. Reprod*.,14, 1328–

SL., Wold BJ. & Pachter L. (2010) *Transcript assembly and quantification by RNA-Seq* 

*reveals unannotated transcripts and isoform switching during cell differentiation.* Nat Biotechnol. 2010 May;28(5):511-5. Epub 2010 May 2.


**3** 

*India* 

Raghava Varman Thampan

**Evolving Trends in Estrogen Receptor Biology** 

Discussions on receptors involved in estrogen action(Stanišić et al,2010) have so far focused on the two major forms of "classical" estrogen receptors, the estrogen receptor α (ER α) and estrogen receptor β (ER β). Both are DNA binding as well as hormone binding forms, with distinct, well-characterized functional domains. The differences between the two have mainly been in their respective molecular masses and shapes and in the target genes with

I have been involved in research in estrogen action for 4 decades and more. Since the focus of my work was chiefly on non-conventional estrogen receptors and receptor associated proteins, often the progress made was felt by me as slow.Nevertheless,it is my sincere belief that what has been unveiled in this direction over the years have not gone unproductive. The two proteins that have been identified in this context, one a non DNA binding estrogen receptor and the other a transcription factor that dimerises with this receptor in the nucleus, have pointed towards the existence of a unique system of receptor in estrogen action. For the first time ever, it has become clear that there is a form of estrogen receptor whose primary functional role is in post transcriptional regulatory mechanisms that include splicing, nucleocytoplasmic transport of RNA and finally, the translation of mRNA. Also, deeper insights into the functional biology of the transcription factor have unfolded certain experimental data hitherto unknown in the literature on steroid hormone action. What is being discussed in this chapter deals exclusively with these two proteins, one a plasma membrane localized estrogen receptor which moves into the nucleus to involve itself in gene regulatory events and the other a transcription factor with a parallel functional role in

The favorite theme of the 60's and early 70's in descriptions of steroid hormone action, particularly with reference to estrogen action, used to be that the receptor primarily existed in the cytosol. Upon hormone binding and the consequent "receptor activation" the receptor entered the nucleus and interacted with the genetic elements. This "two-step mechanism", independently proposed by the research groups led by Jensen and Gorski (Jensen & DeSombre, 1973; Shyamala & Gorski, 1969; Mohla et al, 1972) formed the basis for all subsequent discussions on intracellular movements of the receptor –steroid hormone

**1. Introduction** 

which they interacted.

mitochondrial steroidogenesis

**2. The concept of steroid hormone receptor activation** 

*MIMS Research Foundation, A Division of Malabar Institute* 

*of Medical Sciences, Ltd. (MIMS), Calicut, Kerala,* 

Wu, Y.L., Yang, X., Ren, Z., McDonnell, D.P., Norris, J.D., Willson, T.M. & Greene, G.L. (2005*) Structural basis for an unexpected mode of SERM-mediated ER antagonism.* Mol Cell 2005, 18:413–424.
