**6. Research by our group**

The research work aimed to identify polymorphisms of candidate genes which increase susceptibility to endometriosis by genetic and molecular methods in 106 women who were

Genetic Polymorphisms and Molecular Pathogenesis of Endometriosis 143

temperature for each primer set. DD RT-PCR allows identification of differentially expressed genes in various cell types and under defined physiologic conditions. DD was performed by the modified method of Hasan et al (2000). PCR was performed using selected primers to study the differential expression of the selected genes in the ectopic and eutopic

*cDNA DD PCR:* 5 sets of random exonic primers were used. DD-PCR was repeated with a

A non-parametric statistic is opted because (1) there is no '*a priori reason*' to assume a particular disease model and (2) the assignment of the status '*unaffected*' is problematic

Allele and genotype frequencies were compared in the patient and control groups. The odds ratio (OR) was used to measure the strength of the association between the frequencies of allele and genotype and endometriosis. The software MedCalc (version 7.4.3.0) was used for statistical analyses. All 'p' values two-tailed and 95% Confidence Intervals (CI) were

\* MW T1T1 T1T2

 \*MW- Molecular Weight ladder Fig. 1. Photograph of a 2% agarose gel stained with ethidium bromide to resolve the 306 base pair intron G insertion polymorphism of the progesterone receptor gene. The 149-base pair band represents the wild type allele (allele T1) and the 455-base pair band represents the mutant allele (allele T2). Lane 1 indicates the molecular weight markers; lanes 2 and 3 show the homozygous wild type T1T1 and the heterozygous T1T2 patterns. There is no

455 bp p

149 bp

different set of seven primers, which were selected arbitrarily.

because a surgical procedure is required to exclude endometriosis.

**7. Results and statistical analysis** 

endometrial tissues.

calculated.

**7.1 PROGINS analysis** 

homozygous mutant T2T2 pattern.

confirmed with the diagnosis of endometriosis and with age matched 140 normal women as controls from South India. Progesterone and Androgen Hormone Receptor *gene polymorphisms*; genomic instability at ectopic and eutopic endometrial tissues resulting in *Micro Satellite Instability* (MSI); *gene expression* in ectopic endometrial tissue by Differential Display Reverse Transcriptase Polymerase Chain Reaction (DD-RT-PCR) to identify novel genes were studied. DNA was isolated from peripheral blood and from fresh tissue samples of both ectopic and eutopic endometrium. We subsequently assessed the polymorphisms of Estrogen Receptor, TNF-alpha and TLR 4 genes. The ongoing research work involves ICAM, IL-6, NOD-2, MMP-2,and MMP-9 genes polymorphism studies.
