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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 被引量：2

Effects of PIK3CB gene expression downregulation by RNA interference on the proliferation and apoptosis of colon cancer cell line HCT116
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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。 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 assay and JC-1 staining. Results： The expressions of PIK3CB gene and its protein PI3Kp110β were downregulated in HCT116 cells transfected with PIK3CB-siRNA. PIK3CB-siRNA-179 was the most effective, and it led to the highest inhibition rate （80%） of HCT116 cell proliferation. At 24,48 and 72 h after transfection, PIK3CB-siRNA-179 group showed an obvious proliferation arrest compared with the negative control group detected by CCK-8 assay. The mitochondrial membrane potential displayed an obvious decrease detected by JC-1 fluorescence staining at 48 h after transfection. Conclusion： Downregulation of PI3Kpl110 β protein expression can depress the proliferation and induce apoptosis of colon cancer cell line HCT116. PIK3CB gene may become the new target of gene therapy for colorectal cancer.
作者 倪丽 宋于刚
机构地区 南方医科大学南方医院消化科
出处 《肿瘤》 CAS CSCD 北大核心 2011年第1期22-27,共6页 Tumor
基金 广东省211工程重点学科建设专项基金(编号:C1030379)
关键词 结直肠肿瘤 1-磷脂酰肌醇-3激酶 p110 Β亚单位 RNA干扰 细胞增殖 细胞凋亡 Colorectal neoplasms 1-Phosphatidylinasitol 3-kinase, p110β subunit RNA interference Cellproliferation Apoptosis
分类号 R735.34 [医药卫生—肿瘤]
  • 相关文献

参考文献19

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二级参考文献21

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共引文献27

同被引文献20

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二级引证文献8

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2011年 第1期

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