The effects of DNA methylation and epigenetic factors on the expression of CD133 in ovarian cancers

Kyung Jin Min, Kyeong A. So, Yung Taek Ouh, Jin-Hwa Hong, Jae Kwan Lee

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background: 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. Methods. Ovarian 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. Results: CD133 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. Conclusions: The 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.

Original languageEnglish
Article number28
JournalJournal of Ovarian Research
Volume5
Issue number1
DOIs
Publication statusPublished - 2012 Oct 17

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trichostatin A
DNA Methylation
Epigenomics
Ovarian Neoplasms
Methylation
Cell Line
decitabine
Endometrial Neoplasms
Polymerase Chain Reaction
Messenger RNA
Neoplastic Stem Cells
Reverse Transcription
Real-Time Polymerase Chain Reaction
Western Blotting
Neoplasms
Proteins

Keywords

  • 5'-aza-2'-deoxycytidine
  • Cancer stem cell
  • CD133
  • Epigenetics
  • Trichostatin A

ASJC Scopus subject areas

  • Oncology
  • Obstetrics and Gynaecology

Cite this

The effects of DNA methylation and epigenetic factors on the expression of CD133 in ovarian cancers. / Min, Kyung Jin; So, Kyeong A.; Ouh, Yung Taek; Hong, Jin-Hwa; Lee, Jae Kwan.

In: Journal of Ovarian Research, Vol. 5, No. 1, 28, 17.10.2012.

Research output: Contribution to journalArticle

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abstract = "Background: 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. Methods. Ovarian 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. Results: CD133 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. Conclusions: The 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.",
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AU - Lee, Jae Kwan

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N2 - Background: 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. Methods. Ovarian 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. Results: CD133 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. Conclusions: The 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.

AB - Background: 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. Methods. Ovarian 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. Results: CD133 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. Conclusions: The 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.

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