CAJ | 학술논문

目的：探讨应用RNA干扰技术沉默 HBx 对 HepG2．2．15细胞中hTERT基因表达的影响。方法（1）将pSIHBV／X质粒转染 HepG2．2．15细胞，RT-PCR法评估沉默效率。（2）MTT法检测转染24 h、48 h、72 h后细胞增殖情况。（3）Real-time PCR和Western blot检测hTERT 表达情况。结果测序结果显示 pSIHBV／X 质粒构建正确；RT-PCR检测 HBV x 基因的沉默效率为53．6％；M T T检测结果显示转染24 h、48 h、72 h后 HepG2．2．15细胞的增殖受抑制，与对照组比较差异有统计学意义（P＜0．05）；Real-time PCR和Western blot检测结果均显示，转染pSIHBV／X质粒后 HepG2．2．15细胞中hTERT基因的mRNA水平和蛋白水平表达分别有不同程度的下调，与对照组比较差异有统计学意义（P＜0．05）。结论 siRNA沉默 HBx基因可抑制 HepG2．2．15细胞中癌症标志基因hT ERT的表达。
목적：탐토응용RNA간우기술침묵 HBx 대 HepG2．2．15세포중hTERT기인표체적영향。방법（1）장pSIHBV／X질립전염 HepG2．2．15세포，RT-PCR법평고침묵효솔。（2）MTT법검측전염24 h、48 h、72 h후세포증식정황。（3）Real-time PCR화Western blot검측hTERT 표체정황。결과측서결과현시 pSIHBV／X 질립구건정학；RT-PCR검측 HBV x 기인적침묵효솔위53．6％；M T T검측결과현시전염24 h、48 h、72 h후 HepG2．2．15세포적증식수억제，여대조조비교차이유통계학의의（P＜0．05）；Real-time PCR화Western blot검측결과균현시，전염pSIHBV／X질립후 HepG2．2．15세포중hTERT기인적mRNA수평화단백수평표체분별유불동정도적하조，여대조조비교차이유통계학의의（P＜0．05）。결론 siRNA침묵 HBx기인가억제 HepG2．2．15세포중암증표지기인hT ERT적표체。
Objective To observe the effect of silencing HBx by RNA interference on the expres-sion of hTERT in HepG2 .2 .15 cells .Methods The pSIHBV/X plasmid was transfected into HepG 2 .2 .15 cells ,RT-PCR was used to evaluate the efficaey of transfection .MTT was conducted to check the cell proliferation afer transfection 24 h ,48 h ,72 h .The expression of hTERT assayed by Real-time PCR and Western blot .Results The results of sequencing indicated that the recombinant plasmid pSIHBV/X sequence was correct .Efficient RNA interference of HBx gene was 53 .6% .The cell pro-liferation was inhibited after 24 h ,48 h ,and 72 h of the transfection ,and there were significantly differences between pSIHBV/X group and control (P<0 .05) .The results of real time PCR and west-ern blot analysis showed that the mRNA and protein expression of hTERT was decreased after trans-fection .Compared with control groups ,hTERT mRNA and protein in test groups was significantly lower (P<0 .05) .Conclusion Silencing HBx gene by siRNA can inhibit the expression of cancer mark-er gene hT ERT in HepG2 .2 .15 cell .