CXC类趋化因子受体4基因RNA干扰对人肝癌细胞HepG2和SMMC-7721增殖和侵袭的影响  

Effects of CXC chemokines receptor 4 gene RNA interference on the proliferation and invasion of human hepatoma cells Hep G2 and SMMC-7721

作　　者：张海涛[1] 覃岭[1] ZHANG Hai-tao;QIN Ling(Hepatobiliary Surgery and Liver Transplantation Center,Beijing You’an Hospital,Capital Medical University,Beijing 100069,China)

机构地区：[1]首都医科大学附属北京佑安医院肝胆外科暨肝移植中心

出　　处：《中国肝脏病杂志（电子版）》2019年第4期61-66,共6页Chinese Journal of Liver Diseases：Electronic Version

基　　金：北京市医院管理局临床医学发展专项经费资助（XMLX201711）

摘　　要：目的利用RNA干扰(RNA interference,siRNA)技术抑制人肝癌细胞HepG2和SMMC-7721中CXC类趋化因子受体4(CXC chemokines receptor 4,CXCR4)基因的表达,探讨其在肝癌细胞体外增殖和侵袭过程中的作用及部分机制。方法设计合成CXCR4特异性siRNA,转染人肝癌细胞HepG2和SMMC-7721,siRNA组与对照组采用脂质体转染法转染CXCR4-siRNA与siRNA-对照,空白组以等剂量生理盐水代替。采用逆转录聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)验证siRNA效果,采用MTT法、流式细胞仪及Transwell小室检测细胞增殖、细胞周期和细胞侵袭情况,采用蛋白质免疫印迹法检测细胞中基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)与血管内皮生长因子(vascular endothelial growth factor,VEGF)蛋白的表达。结果 siRNA组Hep G2细胞和SMMC-7721细胞的增殖率分别为0.45±0.12和0.42±0.03,对照组为1.02±0.44和1.07±0.51,空白组分别为1.08±0.44和1.11±0.08,si RNA组细胞增殖率均显著低于对照组和空白组(P均<0.05)。si RNA组Hep G2细胞和SMMC-7721细胞的S期与G2/M期比例显著高于对照组和空白组,G0/G1期比例显著低于对照组和空白组,差异有统计学意义(P均<0.05)。si RNA组Hep G2细胞和SMMC-7721细胞的侵袭数分别为(4.55±1.49)个和(2.48±0.48)个,显著低于对照组[(42.48±3.18)个和(42.00±2.78)个]和空白组[(38.27±2.47)个和(38.19±3.11)个],差异有统计学意义(P均<0.05)。si RNA组Hep G2细胞MMP-9与VEGF蛋白表达量分别为2.09±0.13和0.78±0.12,SMMC-7721细胞MMP-9与VEGF蛋白表达量分别为2.11±0.14和0.81±0.13,均显著低于对照组和空白组(P均<0.05)。结论 RNA干扰CXCR4基因的表达可抑制人肝癌细胞Hep G2和SMMC-7721的增殖和侵袭,其可能是通过调控MMP-9及VEGF蛋白的表达来调节肝癌细胞的增殖和侵袭。Objective RNA interference(siRNA) technology was used to inhibit the expression of CXC chemokines receptor 4(CXCR4) gene in human hepatocellular carcinoma cells HepG2 and SMMC-7721 to explore the role and mechanism of CXCR4 gene in the proliferation and invasion of hepatoma cells in vitro. Methods CXCR4 specific si RNA were designed and synthesized, and were transfected to the human hepatoma cells HepG2 and SMMC-7721. The siRNA group and the control group were transfected with CXCR4-siRNA and siRNA-control by liposome transfection. The blank group was set with an equal dose of normal saline. The effects of siRNA was verified by reverse transcription-polymerase chain reaction(RT-PCR). Cell proliferation, cell cycle and cell invasion were detected by MTT, flow cytometry, Transwell assay. The matrix metalloproteinase-9(MMP-9) and vascular endothelial growth factor(VEGF) expression were detected by Western blot. Results The cell proliferation rates of HepG2 and SMMC-7721 in siRNA group were 0.45 ± 0.12 and 0.42 ± 0.03, respectively, which were 1.02 ± 0.44 and 1.07 ± 0.51 in control group, respectively, and 1.08 ± 0.44 and 1.11 ± 0.08 in blank group, respectively. The cell proliferation rate in siRNA group were significantly lower than those in control group and blank group(all P < 0.05). Compared with control group and blank group, the proportion of S phase and G2/M phase in siRNA group increased significantly, and the proportion of G0/G1 phase decreased significantly, the differences were statistically significant(all P < 0.05). The number of cell in vasion of HepG2 and SMMC-7721 in siRNA group were 4.55 ± 1.49 and 2.48 ± 0.48, respectively, which were significantly lower than those in control group(42.48 ± 3.18 and 42.00 ± 2.78, respectively)and blank group(38.27 ± 2.47 and 38.19 ± 3.11, respectively), the differences were statistically significant(all P < 0.05). The expression of MMP-9 and VEGF in HepG2 in siRNA group were 2.09 ± 0.13 and 0.78 ± 0.12, respectively, and in SMMC-7721 were 2.11 ± 0.14

关 键 词：CXC类趋化因子受体4 RNA干扰 肝癌 增殖 侵袭 

分 类 号：R73[医药卫生—肿瘤]