Abstract
BackgroundTo identifying the effects of DNA methylation and epigenetic factors on the expression of CD133, a cancer stem cell marker, in gynecologic cancer cell lines.MethodsOvarian cancer cell lines (OVCAR-8 and IGROV-1) and an endometrial cancer cell line (Ishikawa) were treated with 5-aza-2`-deoxycytidine (DAC) or Trichostatin A (TSA). Expression of CD133 was evaluated by quantitative real-time PCR, methylation-specific PCR (MSP), reverse transcription-PCR, western blot, and FACS analysis. All results are representative of three independent experiments.ResultsCD133 mRNA expression varied among the different cell lines; the weakest expression was observed in OVCAR-8 cells, while it was strongly expressed in Ishikawa cells. The degree of methylation of the CD133 P2 promoter was 61% in OVCAR-8 cells, 53% in IGROV-1 cells, and 43% in Ishikawa cells. CD133 expression was increased at both the mRNA and protein level after DAC treatment. On the contrary, CD133 mRNA expression decreased after TSA treatment decreased in all cell lines except OVCAR-8. In addition, MSP of the CD133 P2 promoter revealed that methylation was reduced after treatment with either DAC or TSA.ConclusionsThe expression of the CD133 antigen in primary ovarian and endometrial cancer cell lines is regulated by epigenetics, as indicated by its increased expression following DAC treatment and irregular expression pattern followed by TSA treatment. In addition, the expression of CD133 was negatively correlated with the degree of methylation of the CD133 P2 promoter.
Highlights
To identifying the effects of DNA methylation and epigenetic factors on the expression of CD133, a cancer stem cell marker, in gynecologic cancer cell lines
CD133 expression was examined in ovarian cancer cell lines (OVCAR8 and IGROV-1) and Ishikawa cells and normalized to GAPDH expression
Each of these cell lines is of an adenocarcinoma origin, the CD133 mRNA expression varied significantly among the cell lines, with the weakest expression observed in OVCAR-8 cells and the strongest expression in Ishikawa cells (Figure 2)
Summary
To identifying the effects of DNA methylation and epigenetic factors on the expression of CD133, a cancer stem cell marker, in gynecologic cancer cell lines. Ovarian cancer is one of the most fatal gynecological cancers. Chemotherapy is administered to ovarian cancer patients after surgery, as surgical treatment does not confer a sufficient treatment effect. Chemotherapy, is not very effective either; ovarian cancer recurs in Interest in the relationship of cancer stem cells and their role in the response to treatment of ovarian cancer is on the rise. Cancer stem cells have specific genetic variations that give them the capability to limitlessly divide and proliferate, like other stem cells, in addition to the continuous production of various cancer cells. As a result of these capabilities, cancer stem cells can mediate cancer occurrence, tolerance to treatment, and recurrence [3]
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