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为探讨结肠癌细胞SW480和HT-29中LHX6基因启动子区CpG岛的甲基化状态,分析去甲基化与结肠癌细胞增殖的关系,用甲基化特异性PCR对SW480和HT-29细胞的LHX6基因启动子区域CpG岛进行检测,对比经5-氮杂-2-脱氧胞苷(5-aza-2-deoxycytidine,5-Aza-CdR)处理后两株细胞该区域甲基化水平之间的差异,用(3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐,噻唑蓝)(MTT)法检测5-Aza-CdR对其增殖的影响,观察5-Aza-CdR对细胞形态的影响.结果表明,SW480与HT-29细胞的LHX6基因启动子区域CpG岛均存在甲基化,分别为41.9%和28.6%;经5-Aza-CdR处理后,SW480甲基化率由41.4%降低到39.7%,HT-29甲基化率由27.4%降低至18.5%(p<0.05);两株细胞的增殖明显被抑制.结肠癌细胞系SW480和HT-29的LHX6基因均存在不同程度的甲基化,研究结果为LHX6可能作为结肠癌肿瘤检测的新分子靶标奠定基础.
To investigate the methylation status of CpG islands in promoter region of LHX6 gene in colon cancer cells SW480 and HT-29, the relationship between demethylation and colon cancer cell proliferation was analyzed. Methylation-specific PCR was used to detect the expression of SW480 and HT-29 The CpG islands of LHX6 gene promoter region were detected in this study. The methylation levels of these two cells in 5-Aza-2-deoxycytidine (5-Aza-CdR) 5-Aza-CdR was detected by MTT assay with (3- (4,5-dimethylthiazol-2) -2,5-diphenyltetrazolium bromide and thiazolyl blue) Aza-CdR on cell morphology was observed.The results showed that the methylation of LHX6 promoter region CpG islands in SW480 and HT-29 cells were 41.9% and 28.6% After Aza-CdR treatment, the methylation rate of SW480 decreased from 41.4% to 39.7%, and the methylation rate of HT-29 decreased from 27.4% to 18.5% (p <0.05). The proliferation of the two cell lines was significantly inhibited. The LHX6 gene of SW480 and HT-29 cell lines all have different degrees of methylation. The results indicated that LHX6 may lay a foundation for the new molecular target of colon cancer detection.