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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。
To investigate the effect of shRNA targeting p21 on apoptosis induced by curcumin in Huh7 hepatocellular carcinoma cells. The effect of curcumin on the expression of p21 mRNA and protein in Huh7 cells was detected by RT-PCR and Western blotting. ShRNA expression vector pSi-p21 targeting p21 was constructed. The expression of p21 was detected by RT-PCR and Western blotting. The effect of pSi-p21 on the apoptosis of Huh7 cells induced by curcumin was detected by DAPI nuclear staining. The experimental results show that curcumin can significantly up-regulate the expression of p21 mRNA and protein. The interference vector pSi-p21 can significantly reduce the expression of p21 and curcumin-induced apoptosis of Huh7 cells. The results suggest that curcumin induces apoptosis of Huh7 cells by up-regulating p21 expression.