姜黄素对皮肤鳞状细胞癌SCL-1细胞的去甲基化作用研究

Study on the demethylation effect of curcumin on skin squamous cell carcinoma SCL-1 cell

目的观察姜黄素作用于皮肤鳞状细胞癌SCL-1细胞后E-cadherin和p14ARF基因甲基化状态及基因表达的改变情况,为姜黄素的临床应用提供理论依据。方法用不同浓度(10μmol/L、20μmol/L、40μmol/L)的姜黄素处理皮肤鳞癌细胞株SCL-1,MTT法检测细胞增殖情况,甲基化特异性PCR(MSP)法检测E-cadherin和p14ARF基因甲基化状态的改变,RT-PCR法检测E-cadherin和p14ARF基因的表达情况。结果姜黄素可抑制SCL-1细胞的增殖,且呈现时间-剂量依懒性;当姜黄素浓度增至40μmol/L时,SCL-1细胞的E-cadherin和p14ARF基因甲基化减弱、表达增强。结论姜黄素可明显抑制SCL-1细胞增殖,适当浓度的姜黄素对SCL-1细胞的E-cadherin和p14ARF基因有一定的去甲基化作用,并可以调控E-cadherin和p14ARF基因的表达。

Objective To observe the methylation status and the changes in gene expression of E- cadherin and P14 ARF gene when SCL- 1 cells were treated with curcunmin,so as to provide the basis for clinical application of curcumin. Methods Skin squamous carcinoma SCL- 1 cells were treated with curcunmin at various dosages（ 10 μmol / L,20 μmol / L and 40 μmol / L）.The cytotoxic effects of SCL- 1cells were measured by MTT assay. Methylation specific PCR（ MSP） was used to detect the methylation status of P14 ARF and E- cadherin gene. The expression of P14 ARF and E- cadherin gene in SCL- 1 cells were detected by RT- PCR assay. Results The proliferation of SCL- 1 cells can be inhibited by curcumin,showing correlation with time and dose（ P〈0. 01）. When the concentration of curcumin was more than 10 μmol / L,the E- cadherin and P14 ARF genes of SCL- 1 cells＇ s methylation decreased and its expression increased. Conclusion The proliferation of SCL- 1 cells can be significantly inhibited with curcumin. The proper concentrations of curcumin can reverse methylation effect on the P14 ARF and E- cadherin gene of SCL- 1 cells,and it can regulate the expressions of E- cadherin and P14 ARF gene.