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目的:研究去甲二氢愈创木酸(nordihydroguaiaretic acid,NDGA)对宫颈癌SiHa细胞增殖的影响及作用机制。方法:MTT法检测NDGA对SiHa细胞生长的影响,FCM法检测NDGA对SiHa细胞周期及凋亡的影响,RT-PCR检测NDGA对p21基因转录的影响,Western印迹法检测NDGA对组蛋白H3总乙酰化水平的影响,染色质免疫沉淀(chromatin immunopre-cipitation,ChIP)-PCR法检测NDGA对p21基因近启动子区域组蛋白H3乙酰化的影响。结果:NDGA能明显抑制SiHa细胞的生长,且呈时间和剂量依赖性;可使SiHa细胞的细胞周期阻滞于G1期,而凋亡率却下降;能明显提高p21基因的转录水平和组蛋白H3的总乙酰化水平,ChIP-PCR检测结果表明NDGA可明显促进p21基因近启动子区域组蛋白H3的乙酰化水平。结论:NDGA促进p21基因近启动子区域及细胞内总体组蛋白H3的乙酰化可能是其抑制宫颈癌SiHa细胞生长的潜在机制。
Objective: To study the effect and mechanism of nordihydroguaiaretic acid (NDGA) on the proliferation of cervical cancer SiHa cells. Methods: The effects of NDGA on the growth of SiHa cells were detected by MTT assay. The effects of NDGA on the cell cycle and apoptosis of SiHa cells were detected by FCM. The transcription of p21 gene was detected by RT-PCR. The effect of NDGA on histone H3 acetylation in the promoter region of p21 gene was examined by chromatin immunopre-cipitation (ChIP) -PCR. Results: NDGA could significantly inhibit the growth of SiHa cells in a time and dose-dependent manner. The cell cycle of SiHa cells was blocked in G1 phase but the apoptosis rate was decreased. The transcription level of p21 gene and histone protein H3 total acetylation levels, ChIP-PCR test results showed that NDGA can significantly promote p21 gene promoter region near the histone H3 acetylation levels. CONCLUSION: NDGA can promote the acetylation of p21 near-promoter region and intracellular total histone H3, which may be a potential mechanism of its inhibition on the growth of SiHa cells.