靶向p21的shRNA对姜黄素诱导肝癌细胞Huh7凋亡的影响

王伟章  张碧鱼  梅文杰  毛建文  李  明

靶向p21的shRNA对姜黄素诱导肝癌细胞Huh7凋亡的影响

王伟章张碧鱼梅文杰毛建文李明

Effect of p21-targeted shRNA on curcumin-induced apoptosis of human hepatoma Huh7 cells

WANG Wei-Zhang

摘要

摘要:
探讨靶向p21的shRNA对姜黄素诱导肝癌细胞Huh7凋亡的影响。采用RT-PCR和Western blotting方法检测姜黄素对肝癌细胞Huh7中p21 mRNA和蛋白表达的影响; 构建针对p21的shRNA干扰表达载体pSi-p21; 采用RT-PCR和Western blotting方法检测pSi-p21对p21 mRNA和蛋白表达的影响; 通过DAPI核染色法检测pSi-p21抑制p21表达后姜黄素诱导Huh7细胞的凋亡情况。实验结果显示, 姜黄素能明显上调p21 mRNA和蛋白的表达水平。干扰载体pSi-p21可以显著降低p21的表达水平和抑制姜黄素诱导肝癌细胞Huh7凋亡。研究结果提示, 姜黄素通过上调p21的表达进而诱导肝癌细胞Huh7发生凋亡。

Abstract:
In the present study, shRNA plasmid of pSi-p21 targeting p21 mRNA was constructed and the effect of p21 shRNA on curcumin-induced apoptosis of human hepatoma Huh7 cells was investigated. The effect of curcumin on the expression of p21 mRNA and protein and the silence efficiency of pSi-p21 were detected with RT-PCR and Western blotting. The effect of pSi-p21 on curcumin-induced apoptosis of Huh7 cells was evaluated with DAPI staining. The results showed that curcumin significantly upregulated p21 mRNA and protein expression, which was knocked down by pSi-p21 of Huh7 cells. DAPI staining results showed that pSi-p21 significantly decreased curcumin-induced apoptosis of Huh7 cells. The data suggested that curcumin induced apoptosis of Huh7 cells via upregulation of p21 expression.

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