**4. References**

352 Breast Cancer – Focusing Tumor Microenvironment, Stem Cells and Metastasis

MCF-7 treated cells (Elahian et al., 2009). Cotreatment with different concentrations of mitoxantrone and dexamethasone increased the sensitivity of MCF-7 and MCF-7/MX cells to the toxic effects of mitoxantrone. In addition, the flow cytometry results showed that dexamethasone could inhibit the efflux and consequently caused increase in the accumulation of mitoxantrone in MCF-7/MX cells. However, ABCG2 inhibition by

These studies also confirmed that suppression role of dexamethasone on ABCG2 expression in MCF-7/MX cells was more significant than MCF-7 cells. It could be a confirmation for higher level of ABCG2 in MCF-7/MX cells compared with their parental cells and also confirmed other studies that show hormonal regulation of MDR gene expression is cell type

In this review we aimed to focuse on the explanation the role of inflammation on the ABCG2 expression and function, using MCF-7 human breast carcinoma cell line. Proinflammatory cytokines have been found to be present within the micro-environment of tumors and inflammation. They are able to modulate the expression and function of different drug transporters. Mosaffa et al. showed that that proinflammatory cytokines IL-1β and TNF-α induce ABCG2 mRNA and protein expression and increase its function in MCF-7 cells. In MCF-7/MX, these cytokines increased ABCG2 protein expression and function,

Cyclooxygenase-2 (COX-2) is induced by mitogenic and inflammatory stimuli such as growth factors and cytokines, which results in enhanced synthesis of PGs in neoplastic and inflamed tissues. Kalalinia et al. studies had aimed to explore the potential link between COX-2 expression and development of multidrug resistance phenotype in MCF-7 cell line. They reported that COX-2 inducer TPA (12-O-tetradecanoylphorbol-13-acetate) caused a considerable increase up to 9-fold in ABCG2 mRNA expression in parental MCF-7 cells, while a slight increase in ABCG2 expression was observed in the resistant cell line MCF-7/MX. They also showed a positive corrolation between ABCG2 protein expression and COX-2 protein level in each cell line. On the other hand, celecoxib (a selective inhibitor of COX-2) up-regulated the expression of ABCG2 mRNA in MCF-7 and MCF-7/MX cells, which was accompanied by increased ABCG2 protein expression. Furthermore, TPA could increase ABCG2 function in all cell lines with the greatest stimulatory effects in MCF-7/MX (more than 6 times the control level). In addition, celecoxib inverted the effects of TPA on

Several studies have demonstrated that anti-inflammatory drugs like NSAIDs and some glucocorticoids could be effective in chemosensitizing of the many carcinoma cell lines to cytotoxic agents. The pharmacological modulation of ABCG2 in MCF-7 cells by dexamethasone and indomethacin was investigated by elahian et al. . They showed that dexamethasone induced downregulation of ABCG2 mRNA compared to controls in both MCF-7 and MCF-7/MX cell lines, whereas no changes were noted in the presence of indomethacin. The level of ABCG2 protein was decreased in dexamethasone treated MCF-7/MX cells. Cotreatment of mitoxantrone with different concentrations of dexamethasone and indomethacin sensitized parental and resistant MCF-7 cells to mitoxantrone cytotoxicity. Dexamethasone also increased the accumulation of mitoxantrone in the MCF-

dexamethasone was not significant in MCF-7 cells (Elahian et al., 2010).

specific (Demeule et al., 1999; Imai et al., 2005).

but they have no influence on the transporter mRNA levels.

ABCG2 function. This effect was more obvious in MCF-7/MX.

7/MX cell line, indicating an inhibitiory effect on the ABCG2 protein.

**3. Conclusion** 


MCF-7 Breast Cancer Cell Line, a Model for the Study of the Association

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**17** 

*1,2,3Slovenia 4Germany* 

**Interaction of Alkylphospholipid Formulations** 

Alkylphospholipids have shown promising results in several clinical studies (Mollinedo 2007) and among them Perifosine (octadecyl(1,1-di-methyl-4-piperidinium-4-yl)phosphate, OPP), and miltefosine (hexadecylphosphatidylcholine (HPC)) seems to be most promising for breast cancer therapy (Fichtner, Zeisig et al. 1994). For this type of tumor, an antitumor effect was found only for hormone receptor negative tumors *in vivo*, while no effect was found for receptor positive tumors. The reason for this difference is not yet understood and requires further studies. The exact mechanism of action of alkylphospholipids on the molecular level is still not well known in detail. It is clear that they do not target DNA, but they insert into the plasma membrane and subsequently induce a broad range of biological

Unfortunately, administration of free (micellar) alkylphospholipids results in unwanted side effects, reflected in gastrointestinal toxicity and hemolytic activity, which limits the application of higher doses of alkylphospholipids. To achieve better therapeutic effects of alkylphospholipids *in vivo* with less side effects, different liposomal formulations of alkylphospholipids have been tested and showed diminished hemolytic activity. On the other hand, in most cases, cytotoxic activity of liposomes was also lower as compared to free

For efficient application of liposomes as nanocarriers in breast cancer therapy it is not only necessary to investigate the properties of the nanocarrier, which has to transport the drug to the (target) cell, but also the properties of the target cell. The main difference between Perifosine (OPP) resistent MCF7 cells and OPP sensitive MT-3 cells is in the uptake of OPP liposomes by cells and the transport of OPP across plasma membrane. At physiological temperatures the rate of transfer of OPP across plasma membrane increases to greater extent in OPP resistant MCF7 cells, while the uptake of liposomal OPP formulations is lower for

**1. Introduction** 

effects, ultimately leading to cell death.

alkylphospholipids (Zeisig et al., 1998).

**with Breast Cancer Cells in the Context** 

**of Anticancer Drug Development** 

Tilen Koklic1,2, Rok Podlipec1,2, Janez Mravljak3, Maja Garvas1, Marjeta Šentjurc1 and Reiner Zeisig4

*3Faculty of Pharmacy, University of Ljubljana, Ljubljana 4Max-Delbrück-Centre for Molecular Medicine, Berlin-Buch,* 

*1Jozef Stefan Institute, Ljubljana* 

*2Center of Excellence NAMASTE, Ljubljana* 

