姜黄素对皮肤鳞癌SCL-1细胞p16INK4a基因去甲基化作用研究

Study on the curcumin's demethylationon on p16INK4a gene of SCL-1 in skin squamous cancer

目的研究姜黄素作用于SCL-1前后细胞形态、增殖、p16INK4a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。方法用不同剂量姜黄素(5μmol/L、10μmol/L、20μmol/L、40μmol/L、80μmol/L)处理皮肤鳞癌细胞株SCL-1,倒置显微镜观察细胞形态变化,MTT法检测细胞增殖情况,甲基化特异性PCR(MSP)检测p16INK4a基因甲基化状态的改变,反转录PCR法检测p16INK4a基因的表达情况。结果姜黄素能够破坏细胞形态,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>10μmol/L时,SCL-1细胞的p16INK4a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,适当浓度姜黄素对SCL-1细胞p16INK4a基因有一定的去甲基化作用,并可以调控p16INK4a基因的表达。

Objective To study cellular morphology,proliferation,methylation status of p16INK4 a gene and changes of gene expression,as SCL-1 was treated with curcumin,so as to provide theoretical foundation for the clinical application of curcumin. Methods SCL-1 of skin squamous cancer cells were treated with various doses of curcumin（ 5 μmol/L,10 μmol/L,20μmol/L,40 μmol/L,80 μmol/L）. Then inverted microscope was used to observe the cellular morphological changes,cell proliferation was determined by MTT method,methylation specific PCR（ MSP） was used to detect the changes of p16INK4 a gene methylation status,and RT-PCR was performed to detect the expression of p16INK4 a gene. Results Curcumin can destroy the morphology and inhibit the proliferation of SCL-1 with time-dose dependence; when the concentration of curcumin was greater than 10 umol/L,p16INK4 a gene of SCL-1 would decrease methylation and increase expression. Conclusion The proliferation of SCL-1 cells can be significantly inhibited with curcumin. The high concentrations of curcumin had a methylation effect on the p16INK4 a gene of SCL-1 cells. And it also can regulate the expression of p16INK4 a gene.