CAJ | 학술논문

目的探讨沉默乙型肝炎病毒编码X蛋白(HBx)基因后对索拉非尼促人肝癌细胞株PLC凋亡的影响及其机制.方法应用小干扰RNA(siRNA)方法沉默HBx基因,应用实时定量聚合酶链反应(Real-time PCR)检测HBx mRNA表达水平,流式细胞技术检测细胞凋亡,Real-time PCR基因芯片检测HBx-siRNA转染后索拉非尼作用肝癌敏感细胞株PLC丝裂原活化蛋白激酶(MAPK)信号通路基因的表达.结果 HBx特异的siRNA作用后,HBx mRNA表达水平为0.61,与其他对照组比较显著下调(P＜0.05).索拉非尼作用HBx-siRNA干扰后PLC细胞株凋亡表达率为43％,显著高于其他对照组(P＜0.05).索拉非尼作用PLC细胞后,与对照组比较,实验组MAPK信号通路中有13个差异表达基因,其中＞2.0倍的有2个,≤0.5倍的有11个.用siRNA干扰去除HBx基因的影响,经索拉非尼作用PLC细胞后,与对照组比较,实验组MAPK信号通路中有37个差异表达基因,其中＞2.0倍的有1个,≤0.5倍的36个.结论沉默HBx基因增强了索拉非尼促人肝癌细胞株PLC的凋亡作用,可能是沉默HBx基因表达后,扩大和增强了索拉非尼对MAPK信号通路基因的抑制作用.
목적 탐토침묵을형간염병독편마X단백(HBx)기인후대색랍비니촉인간암세포주PLC조망적영향급기궤제.방법 응용소간우RNA(siRNA)방법침묵HBx기인,응용실시정량취합매련반응(Real-time PCR)검측HBx mRNA표체수평,류식세포기술검측세포조망,Real-time PCR기인심편검측HBx-siRNA전염후색랍비니작용간암민감세포주PLC사렬원활화단백격매(MAPK)신호통로기인적표체.결과 HBx특이적siRNA작용후,HBx mRNA표체수평위0.61,여기타대조조비교현저하조(P＜0.05).색랍비니작용HBx-siRNA간우후PLC세포주조망표체솔위43％,현저고우기타대조조(P＜0.05).색랍비니작용PLC세포후,여대조조비교,실험조MAPK신호통로중유13개차이표체기인,기중＞2.0배적유2개,≤0.5배적유11개.용siRNA간우거제HBx기인적영향,경색랍비니작용PLC세포후,여대조조비교,실험조MAPK신호통로중유37개차이표체기인,기중＞2.0배적유1개,≤0.5배적36개.결론 침묵HBx기인증강료색랍비니촉인간암세포주PLC적조망작용,가능시침묵HBx기인표체후,확대화증강료색랍비니대MAPK신호통로기인적억제작용.
Objective To explore whether hepatitis B virus X protein (HBx) gene specific small interfering RNA (siRNA) enhances the effect of sorafenib promoting liver cancer cell line PLC apoptosis and the possible mechanism.Methods PLC cells were treated with HBx-siRNA or in combination with sorafenib.The level of HBx mRNA was assessed by using real-time quantitative polymerase chain reaction (Real-time PCR).The apoptosis rate was detected by using flow cytometry.The changes in genes profiles of mitogen-activated protein kinase (MAPK) signal pathway before and after treatment with HBx-siRNA in combination with sorafenib in PLC cells were observed by using real-time gene array.Results After treatment with HBx-siRNA for 24 h,the expression of HBx mRNA in PLC cells was decreaseed significantly to 0.61 (P ＜ 0.05).HBx-specific siRNA could enhance apoptosis of PLC cells significantly (P ＜ 0.05) and the apoptosis rate was 43％.After treatment with sorafenib in PLC cells,the expression of two genes was increased to more than two times,and that of 11 decreased to below 0.5 times in the MAPK signal pathway by using real-time PCR array.After HBx siRNA and treatment with sorafenib,the expression of one gene was increased to more than two times,and that of 37 decreased to below 0.5 times in the MAPK signal pathway by using real-time PCR array.Conclusion HBx-si RNA suppresses Hbx gene expression and enhances the response of PLC cells to sorafenib.The mechanism is related to more systemic inhibitory effect on MAPK signal pathway.