Effects of cyclin-dependent kinase 8 specific siRNA on the proliferation and apoptosis of colon cancer cells.
Effects of cyclin-dependent kinase 8 specific siRNA on the proliferation and apoptosis of colon cancer cells.
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DOI：
10.1186/1756-9966-30-109
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发表时间：
2011-11-22
期刊：
影响因子：
--
通讯作者：
Zhu XG
中科院分区：
文献类型：
--
作者：
He SB;Yuan Y;Wang L;Yu MJ;Zhu YB;Zhu XG
目的研究细胞周期蛋白依赖性激酶8（cyclin-dependent kinase 8，CDK 8）和β-catenin在结肠癌中的表达，探讨CDK 8在结肠癌细胞尤其是结肠癌细胞株HCT 116增殖、凋亡和细胞周期进程中的作用。用靶向CDK 8的小干扰RNA（siRNA）转染结肠癌细胞系HCT 116。RT-PCR和Western blot检测CDK 8-siRNA转染后HCT 116细胞中CDK 8和β-catenin的mRNA和蛋白表达水平。MTT法检测细胞增殖活性，流式细胞仪（FCM）检测细胞周期和凋亡情况。采用实时荧光定量PCR和免疫组化方法检测结肠癌组织和癌旁正常组织中CDK 8和β-catenin蛋白的表达。CDK 8特异性siRNA转染后，HCT 116细胞中CDK 8和β-catenin的mRNA和蛋白表达水平均明显降低（P < 0.05）。CDK 8特异性siRNA转染能显著抑制HCT 116细胞增殖，促进其凋亡（P < 0.05）。转染CDK 8-siRNA后，HCT 116细胞G 0/G1期细胞比例明显增加（P < 0.05）。CDK 8和β-catenin在癌旁正常组织中的表达低于癌组织（P < 0.05）。CDK 8与β-catenin在胃癌及癌旁正常组织中的表达均呈正相关（P < 0.05）。CDK 8和β-catenin在结肠癌中的表达频率较高。CDK 8特异性siRNA转染可下调结肠癌细胞中CDK 8的表达，并降低β-catenin的表达。此外，CDK 8特异性siRNA可抑制结肠癌细胞的增殖，促进其凋亡，并使结肠癌细胞阻滞于G 0/G1期。干扰CDK 8可能是通过β-catenin调控结肠癌的有效策略。
To investigate the expression of cyclin-dependent kinase 8 (CDK8) and β-catenin in colon cancer and evaluate the role of CDK8 in the proliferation, apoptosis and cell cycle progression of colon cancer cells, especially in HCT116 cell line. Colon cancer cell line HCT116 was transfected with small interfering RNA (siRNA) targeting on CDK8. After CDK8-siRNA transfection, mRNA and protein expression levels of CDK8 and β-catenin were determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot assay in HCT116 cells. Cell proliferation was measured by 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide Methylthiazolyl tetrazolium (MTT) assay, and cell cycle distribution and apoptosis were analyzed by flow cytometry analysis (FACS). CDK8 and β-catenin protein levels were also examined by real-time PCR and immunohistochemistry (IHC) in colon cancer tissues and adjacent normal tissues. After CDK8 specific siRNA transfection, mRNA and protein expression levels of CDK8 and β-catenin in HCT116 cells were noticeably decreased (P < 0.05). CDK8 specific siRNA transfection inhibited HCT116 cells' proliferation and facilitated their apoptosis significantly (P < 0.05). In addition, the proportion of HCT116 cells in the G0/G1 phase was remarkably increased after CDK8-siRNA transfection (P < 0.05). The expression levels of CDK8 and β-catenin in adjacent normal tissues were lower than in tumor tissues (P < 0.05). Moreover, the expression of CDK8 was correlated with the expression of β-catenin in both tumor and adjacent normal tissues (P < 0.05). CDK8 and β-catenin were expressed in colon cancer at a high frequency. CDK8 specific siRNA transfection down-regulated the expression of CDK8 in colon cancer cells, which was also associated with a decrease in the expression of β-catenin Moreover, CDK8 specific siRNA inhibited the proliferation of colon cancer cells, promoted their apoptosis and arrested these cells in the G0/G1 phase. Interference of CDK8 might be an effective strategy through β-catenin regulation of colon cancer.
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