CAJ | 학술논문

目的研究大鼠肝脏卵圆细胞中乙型肝炎病毒编码X蛋白(HBX)对β型转化生长因子(TGFβ)1增殖抑制效应的影响.方法转染HBX基因的大鼠肝脏卵圆细胞系HBX-EGFP-LE/6作为实验组,大鼠肝脏卵圆细胞系LE/6和转染绿色荧光标记空载体的大鼠肝脏卵圆细胞系EGFP-LE/6作为空白对照和阴性对照组.采用real-time PCR和Western blot分别检测各组TGFβ受体2的mRNA及蛋白表达水平.各组细胞在体外培养中加入μg/L外源性TGFβ1,采用MTT增殖实验比较各组细胞对外源性TGFβ1细胞增殖抑制效应的不同反应性.结果实验组TGFβ受体2的mRNA和蛋白表达水平较两个对照组均明显下调(P<0.05).MTT增殖实验结果表明HBX在体外影响TGFβ1对卵圆细胞的增殖抑制作用,稳定转染HBX的卵圆细胞株HBX-EGFP-LE/6对TGFβ1的增殖抑制效应的反应性为18.1%±1.5%,低于其余两组的42.2%±2.8%和41.9%±5.0%(P<0.05).结论 HBX在肝脏卵圆细胞中的表达影响TGFβ受体2在卵圆细胞中的转录活性和蛋白合成,并且降低肝脏卵圆细胞对TGFβ1增殖抑制效应的敏感度.
목적 연구대서간장란원세포중을형간염병독편마X단백(HBX)대β형전화생장인자(TGFβ)1증식억제효응적영향.방법 전염HBX기인적대서간장란원세포계HBX-EGFP-LE/6작위실험조,대서간장란원세포계LE/6화전염록색형광표기공재체적대서간장란원세포계EGFP-LE/6작위공백대조화음성대조조.채용real-time PCR화Western blot분별검측각조TGFβ수체2적mRNA급단백표체수평.각조세포재체외배양중가입μg/L외원성TGFβ1,채용MTT증식실험비교각조세포대외원성TGFβ1세포증식억제효응적불동반응성.결과 실험조TGFβ수체2적mRNA화단백표체수평교량개대조조균명현하조(P<0.05).MTT증식실험결과 표명HBX재체외영향TGFβ1대란원세포적증식억제작용,은정전염HBX적란원세포주HBX-EGFP-LE/6대TGFβ1적증식억제효응적반응성위18.1%±1.5%,저우기여량조적42.2%±2.8%화41.9%±5.0%(P<0.05).결론 HBX재간장란원세포중적표체영향TGFβ수체2재란원세포중적전록활성화단백합성,병차강저간장란원세포대TGFβ1증식억제효응적민감도.
Objective To determine whether hepatitis B virus X (HBX) protein expression affect the oval cells' response to anti-proliferative effect of transforming growth factor β1 (TGFβ1) in oval cells. Methods Real-time PCR, Western blot analysis were performed to detect the expression of TGFpR II in HBX-transfected oval cells named HBX-EGFP-LE/6, and EGFP-LE/6, LE/6 control cells. In addition, exogenous TGFβ1 was added into all three oval cell lines, MTT assay was preformed to clarify different responses to the anti-proliferative effect of TGFβ1. Results The TGFβR II mRNA levels in LE/6 and EGFP-LE/6 cells were (10. 2 ± 1. 8) and (8. 8 ± 0. 9) folds of those in HBX-EGFP-LE/6 cells, the difference was significant (P < 0. 05). HBX protein expression also reduced the protein levels of TGFβR II in HBX-EGFP-LE/6 oval cells, compared to the control cells. The MTT results exhibited that, after TGFβ1 addition, proliferative inhibition rate in the HBX-EGFP-LE/6 cells was 18. 1% ± 1.5% while those in control cells were 42. 2% ± 2. 8% and 41.9% ± 5. 0% , the difference was significant (P < 0. 01). Conclusion HBX protein expression affects TGFβR II transcriptional activity and protein synthesis, and insensitive oval cells to anti-proliferative effect of TGFβ1.