CXCL8 siRNA通过PI3K/Akt/NF-κB信号途径抑制结肠癌细胞的增殖和侵袭  

CXCL8 siRNA inhibits proliferation and invasion of human colon carcinoma cells through PI3K/Akt/NF-κB signaling pathway

作　　者：马家驰[1] 李渊[1] 李一平[1] 房伟[1] 郭庆金 

机构地区：[1]甘肃省人民医院普外科,甘肃兰州730000 [2]宁夏医科大学研究生院,宁夏回族自治区银川750004

出　　处：《肿瘤》2015年第6期604-612,共9页Tumor

基　　金：国家自然科学基金资助项目(编号:81260325);甘肃卫生行业科研计划资助项目(编号:GSWST2010-17)~~

摘　　要：目的:探讨CXC趋化因子配体8[chemokine(C-X-C motif)ligand 8,CXCL 8]基因沉默对结肠癌细胞增殖和侵袭的影响,以及磷脂酰肌醇3激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,PKB,又称Akt)/核因子κB(nuclear factor-kappa B,NF-κB)信号转导途径在其中所起的作用。方法 :应用RT-PCR和蛋白质印迹法检测CXCL8在4种结肠癌细胞株HT-29、Wi Dr、Ca Co-2和Co Lo320中的表达水平。应用Lipofect AMINE2000脂质体转染法将靶向CXCL 8基因的小干扰RNA(small interfering RNA,si RNA)转染至4种结肠癌细胞,然后采用蛋白质印迹法检测CXCL8蛋白的表达。WST-1法和Transwell小室实验分别检测CXCL 8基因沉默对4种结肠癌细胞增殖和侵袭能力的影响。蛋白质印迹法检测CXCL 8基因沉默后HT-29细胞中PI3K/Akt/NF-κB通路主要蛋白的磷酸化水平的变化。结果 :CXCL8在4种结肠癌细胞株中均高表达,而CXCL8 si RNA转染后CXCL8蛋白表达均被明显抑制(P值均<0.01)。转染CXCL8 si RNA后结肠癌细胞的增殖和侵袭能力均明显降低(P值均<0.01)。HT-29细胞中CXCL 8基因沉默可明显抑制CXCL8诱导的PI3K、Akt和NF-κB蛋白磷酸化(P值均<0.01)。结论 :CXCL8基因沉默能够显著抑制结肠癌细胞的增殖和侵袭,推测其机制与下调PI3K/Akt/NF-κB信号通路蛋白的活化相关。Objective： To investigate the effects of chemokine（C-X-C motif） ligand 8（CXCL 8） gene silencing on cell proliferation and invasion of human colon cancer cells, and to investigate its relationship with phosphatidylinositol 3-kinase/protein kinase B/nuclear factor-kappa B（PI3K/Akt/NF-κB） signaling pathway.Methods： The expression levels of CXCL8 mRNA and protein in four human colon cancer HT-29, Wi Dr, Ca Co-2 and Co Lo320 cells were detected by reverse transcription-PCR and Western blotting, respectively. The four colon cancer cells were transfected with the small interfering RNA targeting CXCL 8 gene（CXCL8 si RNA） by Lipofect AMINE 2000, then the expression of CXCL8 protein was detected by Western blotting. The proliferation and invasion abilities of colon cancer cells after CXCL 8 gene silencing were detected by WST-1 method and Transwell invasion assay, respectively. The phosphorylation levels of PI3 K, Akt and NF-κB proteins in HT-29 cells with CXCL8 gene silencing were detected by Western blotting.Results： CXCL8 m RNA and protein were highly expressed in four colon cancer cell lines. After CXCL8 siRNAs were transfected into the colon cancer cells, the expression of CXCL8 protein was significantly inhibited（all P 〈 0.01）, while the proliferation and invasion abilities of colon cancer cells were significantly decreased（all P 〈 0.01）. CXCL 8 gene silencing resulted in blockage of PI3 K, Akt and NF-κB protein phosphorylation induced by CXCL8 in colon cancer HT-29 cells（all P 〈 0.01）.Conclusion： CXCL8 gene silencing significantly inhibits the proliferation and invasion of colon carcinoma cells. It may be related to down-regulation of protein activity in PI3K/Akt/NF-κB signaling pathway.

关 键 词：结肠肿瘤 RNA干扰 白细胞介素8 细胞增殖 肿瘤浸润 PI3K/Akt/NF-κB信号通路 

分 类 号：R735.35[医药卫生—肿瘤]