| RNA干扰靶向抑制PIK3CB基因表达对结肠癌HCT116细胞增殖和凋亡的影响 |
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| 引用本文： | 倪丽,宋于刚.RNA干扰靶向抑制PIK3CB基因表达对结肠癌HCT116细胞增殖和凋亡的影响[J].肿瘤,2011,31(1):22-27. |
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| 作者姓名： | 倪丽 宋于刚 |
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| 作者单位： | 南方医科大学南方医院消化科,广东省胃肠疾病重点实验室,广州,510515 |
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| 基金项目： | 广东省211工程重点学科建设专项基金 |
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| 摘 要： | 目的:探讨RNA干扰靶向抑制PIK3CB基因(编码PI3Kp110β蛋白)的表达对结肠癌HCT116细胞增殖和凋亡的影响。方法:设计3条针对PIK3CB基因的小干扰RNA(small interfering RNA,siRNA)片段及1条阴性对照siRNA,分别瞬时转染结肠癌HCT116细胞。用Real time PCR及Western印迹法筛选出1条抑制效率最高的PIK3CB-siRNA序列,然后进行CCK-8细胞增殖实验,并采用JC-1染色法检测线粒体膜电位(Δψm)的变化。结果:经转染PIK3CB-siRNA-179序列的HCT116细胞中PI3Kp110βmRNA及蛋白表达降低最为明显,抑制率达80%,选择该条siRNA作为有效干扰组进行后续实验。CCK-8细胞增殖实验显示,瞬时转染24、48及72h后,PIK3CB-siRNA-179组细胞的相对存活率较阴性对照组细胞明显降低,其中以24h及48h降低最为明显(P<0.05)。JC-1染色结果显示,瞬时转染48h后PIK3CB-siRNA-179组细胞的线粒体膜电位较阴性对照组细胞明显降低。结论:抑制PI3Kp110β蛋白表达可降低结肠癌HCT116细胞的增殖速度,诱导细胞的凋亡,推测PIK3CB基因可能成为结肠癌基因治疗的一个新靶点。
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| 关 键 词： | 结直肠肿瘤 1-磷脂酰肌醇-3激酶 p110β亚单位 RNA干扰 细胞增殖 细胞凋亡 |
Effects of PIK3CB gene expression downregulation by RNA interference on the proliferation and apoptosis of colon cancer cell line HCT116 |
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| NI Li,SONG Yu-gang.Effects of PIK3CB gene expression downregulation by RNA interference on the proliferation and apoptosis of colon cancer cell line HCT116[J].Tumor,2011,31(1):22-27. |
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| Authors: | NI Li SONG Yu-gang |
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| Institution: | NI Li,SONG Yu-gang Department of Gastroenterology,Guangdong Provincial Key Laboratory of Gastroenterology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China |
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| Abstract: | Objective: To explore the effects of expression downregulation of PIK3CB gene induced by RNA interference on the proliferation and apoptosis of colon cancer HCT116 cells.Methods: Three PIK3CB small interfering RNA (siRNA) sequences and one negative control siRNA sequence were designed and synthesized,and then individually transfected into HCT116 cells via lipofectAMINE2000 mediation.The most effective PIK3CB-siRNA was screened by real time PCR and Western blotting.Then the target siRNA was used for CCK-8 as... |
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