乙肝病毒X蛋白可通过调控CXC趋化因子受体6表达促进肝癌细胞的侵袭和转移

Hepatitis B virus X protein can promote the invasion and metastasis of hepatocellular carcinoma cells by regulating the expression of CXC Chemokine Receptor 6

目的探究乙型肝炎病毒X蛋白(HBx)通过调控CXC趋化因子受体6(CXCR6)对肝癌HepG2细胞生长和凋亡的作用。方法按处理方式不同将HepG2细胞分成Control组、NC组(转染阴性对照质粒)、HBx组(转染HBx mimics)、HBx+si-CXCR6组(共转染HBx-mimics和CXCR6 siRNA)。二苯基四氮唑溴盐(MTT)、流式细胞术、Transwell实验分别检测细胞增殖、凋亡率、迁移和侵袭情况,蛋白质印迹法(Western blot)检测细胞中CXCR6和HBx蛋白表达。多组间比较采用单因素方差分析,组间两两比较采用SNK-q检验。结果HBx组CXCR6、HBx蛋白表达及细胞增殖水平高于Control组和NC组[(0.49±0.05)比(0.27±0.04、0.26±0.03)、(0.59±0.05)比(0.32±0.04、0.30±0.05)、(0.96±0.07)比(0.66±0.03、0.62±0.03),t=5.951、5.732、7.304、8.195、6.823、7.495,P<0.05]。HBx+si-CXCR6组CXCR6蛋白表达及细胞增殖水平低于HBx组(0.18±0.02比0.49±0.05、0.63±0.02比0.96±0.07,t=9.971、7.851,P<0.05)。HBx组细胞增殖水平、迁移率、侵袭数高于Control组和NC组[(0.96±0.07)比(0.66±0.03)、(0.62±0.03)、(46.38±0.06)%比(20.21±0.04)%、(23.23±0.03)%、(78.53±0.06)比(36.19±0.02)、(39.18±0.03),t=7.914、7.833、628.583、614.025、1159.529、1024.361,P<0.05],细胞凋亡率低于Control组和NC组[(1.33±0.16)%比(5.84±0.11)%、(6.63±0.39)%,t=40.232、51.381,P<0.05]。HBx+si-CXCR6组细胞增殖水平、迁移率、侵袭数低于HBx组[(0.63±0.02)比(0.96±0.07)、(19.41±0.05)%比(46.38±0.06)%、(33.51±0.04)%比(78.53±0.06)%,t=8.025、603.548、1120.846,P<0.05],细胞凋亡率高于HBx组[(4.66±0.36)%比(1.33±0.16)%,t=45.715,P<0.05]。结论乙肝病毒X蛋白可通过提高CXCR6表达促进肝癌细胞的侵袭和转移。

Objective To investigate the role of Hepatitis B x protein(HBx)on the growth and apoptosis of hepatocellular carcinoma HepG2 cells through the regulation of CXC Chemokine Receptor 6(CXCR6).Methods HepG2 cells were divided into Control group,NC group(transfected with negative control plasmid),HBx group(transfected with HBx mimics)and HBx+si-CXCR6 group(co-transfected with HBX-Mimics and CXCR6 siRNA)according to different treatments.Cell proliferation,apoptosis,migration and invasion were detected by methyl thiazolyl tetrazolium(MTT),flow cytometry and Transwell assay,respectively.CXCR6 and HBx protein expressions were detected by Western blotting.One-way analysis of variance was used for comparison between groups,and SNK-q test was used for pair-to-group comparison.Results CXCR6 and HBx protein expression and cell proliferation levels in HBx group were higher than those in Control group and NC group[(0.49±0.05)compared to(0.27±0.04,0.26±0.03),(0.59±0.05)compared to(0.32±0.04,0.30±0.05),(0.96±0.07)compared to(0.66±0.03,0.62±0.03),t=5.951,5.732,7.304,8.195,6.823,7.495,P<0.05].The expression of CXCR6 protein and the level of cell proliferation in HBx+si-CXCR6 group were lower than those in HBx group[(0.18±0.02)compared to(0.49±0.05),(0.63±0.02)compared to(0.96±0.07),t=9.971,7.851,P<0.05].The cell proliferation level,migration rate,and invasion number in HBx group were higher than those in Control group and NC group[(0.96±0.07)compared to(0.66±0.03),(0.62±0.03),(46.38±0.06)%compared to(20.21±0.04)%,(23.23±0.03)%,(78.53±0.06)compared to(36.19±0.02),(39.18±0.03),t=7.914,7.833,628.583,614.025,1159.529,1024.361,P<0.05],the apoptosis rate was lower in the Control group than in the NC group[(1.33±0.16)%compared to(5.84±0.11)%,(6.63±0.39)%,t=40.232,51.381,P<0.05].The cell proliferation level,migration rate,and invasion number in HBx+si-CXCR6 group were lower than those in HBx group[(0.63±0.02)compared to(0.96±0.07),(19.41±0.05)%compared to(46.38±0.06)%,(33.51±0.04)%compared to(78.53±0.06)%,t=8.025,603.548,1120.846,P<0.05],and the apoptosis rate was higher than those in HBx group[(4.66±0.36)%compared to(1.33±0.16)%,t=45.715,P<0.05].Conclusion Hepatitis B virus X protein can promote the invasion and metastasis of liver cancer cells by increasing the expression of CXCR6.