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. 2003 Sep;9(9):1976-80.
doi: 10.3748/wjg.v9.i9.1976.

Effects of DNA methylation on expression of tumor suppressor genes and proto-oncogene in human colon cancer cell lines

Jing-Yuan Fang  1 , Juan Lu, Ying-Xuan Chen, Li Yang
Affiliations

Effects of DNA methylation on expression of tumor suppressor genes and proto-oncogene in human colon cancer cell lines

Jing-Yuan Fang et al. World J Gastroenterol. 2003 Sep.

"V体育平台登录" Abstract

Aim: To investigate the effects of DNA methylation on the expression of tumor suppressor genes and proto-oncogene in human colon cancer cell lines. VSports手机版.

Methods: Three colon cancer cell lines (HT-29, SW1116 and Colo-320) treated with different concentrations of DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (5-aza-dC) were used to induce DNA demethylation. The expressions of p16(INK4A), p21(WAF1), APC and c-myc genes were observed by using RT-PCR. The methylation status of p16(INK4A) promoter in HT-29 cells was also determined by methylation-specific PCR (MSP) V体育安卓版. .

Results: Weak expressions of p16(INK4A) and APC in the three colon cancer cells were detected, and p21(WAF1) expression was not found in SW1116 and Colo-320 cells before treatment V体育ios版. After treatment of 1 micromol/L but not 10 micromol/L of 5-aza-dC, the methylation level of p16(INK4A) gene promoter decreased significantly, and the hypomethylation led to the up-regulation of p16(INK4A) gene transcription in HT-29 cells. In the cell lines of SW1116 and Colo-320, p16(INK4A) and APC mRNA expressions were obviously enhanced after treatment of either 10 micromol/L or 5 micromol/L 5-aza-dC for 24 h. However, no evidence was found that methylation regulated the expression of p21(WAF1) and c-myc genes in human colon cancer cell lines. .

Conclusion: Expression of p16(INK4A) and APC genes is regulated by DNA methylation in three human colon cancer cell lines. VSports最新版本.

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Figures

Figure 1
Figure 1
5-aza-dC induced hypomethylation of the promoter of p16INK4A gene in HT-29 cells. Lane 1, untreated; lane 2, 5-aza-dC treated; lane 3, untreated with bisulfite. MSP was performed with the specific primers described in the Materials and Methods.
Figure 2
Figure 2
Up-regulated mRNA level of p16INK4A by 5-aza-dC in HT-29 cells. RT-PCR was performed as described in Materials and Methods. β-actin was used as a loading/amplification control.
Figure 3
Figure 3
5-aza-dC increased the transcription of p16INK4A gene in Colo-320 (A) and SW1116 cells. Lane 1: mock treatment. Lanes 2-7: after 5-aza-dC treatment; lane 2: 2 μmol/L, 24 h; lane 3: 5 μmol/L, 24 h; lane 4: 10 μmol/L, 24 h; lane 5: 2 μmol/L, 72 h; lane 6: 5 μmol/L, 72h; lane 7: 10 μmol/L, 72 h. The density of bands shown in Table 3.
Figure 4
Figure 4
5-aza-dC increased the transcription of APC gene in Colo-320 (A) and SW1116 cells. Lane 1: mock treatment. Lanes 2-7: after 5-aza-dCtreatment; lane 2: 2 μmol/L, 24 h; lane 3: 5 μmol/L, 24 h; lane 4: 10 μmol/L, 24 h; lane 5: 2 μmol/L, 72 h; lane 6: 5 μmol/L, 72 h; lane 7: 10 μmol/L, 72 h. The density of bands shown in Table 4.
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