Objective To investigate the effects of siRNA-target PICK1 interference on proliferation, apoptosis and invasion of human colon adenocarcinoma HT-29 cells. Methods Sequence of small interfering RNA (siRNA) targeting PICK1 was design and synthesized. Three groups were set: experimental group (PICK1-siRNA), the negative control group (negative-siRNA) and the control group. Cationic liposomes (LipofectamineTM2000) were used to medi-ate and transfect the PICK1 gene into HT-29 cells. MTT assay was used to detect the activity of HT-29 cell prolifer-ation. Western-blotting was used to measure the expression of PICK1 protein before and after the transfection. Cell apoptosis was detected by flow cytometry. Transwell cell invasion assay was used to measure the invasion of HT-29 cells. Results At 24h after transfection, western-blotting showed that the relative expression of PICK1 protein in PICK1-siRNA group was lower than those in the negative-siRNA group and control group (both P<0.05). MTT assay showed that the HT-29 proliferation was significantly inhibited in PICK1-siRNA group, with statistically significant-from the other groups (P<0.05). Compared with the negative-siRNA group and control group, the PICK1-siRNA group showed significantly higher proportion of apoptotic HT-29 cells andreduced invasiveness, with statistically significant difference (P<0.05). Conclusion Interfering PICK1 expression may effectively inhibit the proliferation, induce apop-tosis, and reduce invasion-related protein expression of HT-29 cells. Therefore PICK1 can be used as a potential tar-get for treatment of colon cancers.%目的 研究小分子干扰RNA(siRNA)干扰蛋白激酶Cα相互作用蛋白(PICK)1基因对人结肠腺癌HT-29细胞增殖、凋亡及侵袭的影响.方法 设计合成针对PICK1基因的siRNA序列,分为实验组(PICK1-siRNA)、阴性对照组(Negative-siRNA)及对照组.阳离子脂质体(LipofectamineTM 2000)介导转染至HT-29细胞,四甲基偶氮唑蓝(MTT)检测HT-29细胞增殖活性的改变;蛋白印迹法检测转染前后PICK1蛋白的表达;流式细胞术检测细胞凋亡的变化;细胞侵袭实验检测细胞侵袭能力的变化.结果 转染24 h后,PICK1-siRNA组PICK1蛋白的相对表达量低于Negative-siRNA组和对照组(P<0.05).MTT结果显示,PICK1-siRNA组HT-29细胞的增殖活性受到明显的抑制作用,差异具有统计学意义(P<0.05).与Negative-siRNA组和对照组相比,PICK1-siRNA组HT-29细胞凋亡比例显著增高,侵袭能力显著下降,差异具有统计学意义(P<0.05).结论 干扰PICK1基因表达可有效抑制HT-29细胞增殖,诱导凋亡,降低侵袭相关蛋白的表达,可作为结肠癌治疗的潜在靶点.
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