KPNA2基因沉默对人肝癌细胞株 SMMC7721和 Bel7404增殖和侵袭能力的影响

摘要

目的：观察核转运蛋白基因α2（Karyopherin α-2，KPNA2）基因沉默对人肝癌细胞株 SMMC7721和 Bel7404增殖以及侵袭能力的影响。方法将 KPNA2 siRNA 干扰质粒用 LipofectaminTM 2000方法瞬时转染人肝癌细胞株SMMC7721和 Bel7404，转染后48 h 应用蛋白质印迹法检测转染细胞中 KPNA2蛋白表达。采用 MTT 法检测基因沉默细胞增殖能力，采用 Transwell 法检测基因沉默细胞侵袭能力。结果 SMMC7721细胞株对照组 mRNA 为1．02±0．13，高于 siRNA 转染组的0．37±0．07（t＝10．78，P ＜0．01）；肝癌 Bel7404细胞株对照组 mRNA 为1．05±0．17，高于 siRNA 转染组的0．36±0．06（t＝9．38，P ＜0．01）。肝癌 SMMC7721细胞株对照组蛋白定量值为0．96±0．10，高于转染组的0．42±0．05（t＝11．83，P ＜0．01）；肝癌 Bel7404细胞株对照组蛋白定量值为0．93±0．09，高于转染组的0．48±0．06（t ＝10．19， P ＜0．01）。SMMC7721和 Bel7404细胞株在24、48和72 h 时对照组增殖能力高于转染组，且差异有统计学意义（P ＜0．05）。SMMC7721细胞株对照组侵袭能力值为126．20±21．61，高于 siRNA 转染组的51．13±10．2（t ＝7．68，P ＜0．01）；肝癌 Bel7404细胞株对照组侵袭能力值为125．124±8．04，高于 siRNA 转染组的55．20±18．54（t ＝8．48，P ＜0．01）。结论 KPNA2基因沉默可以调节人肝癌细胞株 SMMC7721和 Bel7404的增殖能力和侵袭能力。%Objective To observe the impacts of Karyopherin α-2 (KPNA2 )gene silencing on proliferation and invasion of human hepatocarcinoma cell lines SMMC7721 and Bel7404.Methods Human hepatocarcinoma cell lines SMMC7721 and Bel7404 were transiently transfected with KPNA2 siRNA through adoption of LipofectaminTM 2000. Protein immunoblotting was performed to detect KPNA2 protein expression in transfected cells at 48 hours after transfection.MTT assay was carried out to assess proliferative capability of gene silencing cells,and Transwell assay was used to evaluate their invasion ability.Results Relative mRNA level of KPNA2 in control group of SMMC7721 and Bel7404 was higher than that in siRNA-transfected group (1 .02±0.13 vs.0.37±0.07,t =10.78,P <0.01 ;1 .05 ±0.1 vs.70.36 ± 0.06,t = 9.38,P < 0.01 ,respectively).Relative protein level in control group was higher than that in transfected group (0.96±0.10 vs.0.42±0.05,t=11 .83,P <0.01 ;0.93 ±0.09 vs.0.48±0.06,t=10.19,P <0.01 , respectively).Proliferative capacity of control groups was stronger than that in transfected groups at 24h,48h and 72h respectively,which revealed statistically significant differences (P <0.05 ).Invasive capacity in control group was more powerful than that in siRNA-transected group (126.20±21 .61 vs.51 .13±10.2,t=7.68,P <0.01 ;125.124±8.04 vs. 55.20 ± 18.54,t = 8.48,P <0.01 ,respectively ).Conclusion KPNA2 gene silencing could adjust the proliferative capacity and invasive ability of human hepatocarcinoma cell lines SMMC7721 and Bel7404.