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Possible mechanism for hepatitis B virus X gene to induce apoptosis of hepatocytes.

机译:乙型肝炎病毒X基因诱导肝细胞凋亡的可能机制。

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AIM: To investigate the possible mechanism for HBV X gene to induce apoptosis of hepatocyte HL-7702 cells. METHODS: HBV X gene eukaryon expression vector pcDNA3-X was established and transfected into HL-7702 cells by lipid-mediated transfection, including transient and stable transfection. Positive clones were screened by incubating in the selective medium with 600 mug/mL G418 and named HL-7702/HBV-encoded X protein (HBx) cells. The expressions of Fas/FasL, Bax/Bcl-2, and c-myc mRNA were measured by semi-quantitative RT-PCR in HL-7702/HBx and control group, respectively. RESULTS: RT-PCR analysis confirmed that HBV X gene was transfected into HL-7702 cells successfully. By semi-quantitative RT-PCR analysis, Bax and c-myc mRNA levels in HL-7702/HBx cells of transient transfection were significantly higher than those in control, FasL and c-myc mRNA levels in HL-7702/HBx cells of stable transfection were significantly higher than those in control, whereas the Bcl-2 mRNA levels in HL-7702/HBx cells of transient and stable transfection were significantly lower than those in control. CONCLUSION: HBV X gene may promote the apoptosis of hepatocytes by regulating the expressions of Fas/FasL, Bax/Bcl-2, and c-myc gene in a dose-dependent manner.
机译:目的:探讨HBV X基因诱导肝细胞HL-7702细胞凋亡的可能机制。方法:建立HBV X基因真核表达载体pcDNA3-X,通过脂质介导的转染,包括瞬时和稳定转染,转染到HL-7702细胞中。通过在选择性培养基中与600杯/毫升G418孵育,筛选出阳性克隆,并将其命名为HL-7702 / HBV编码的X蛋白(HBx)细胞。通过半定量RT-PCR分别检测HL-7702 / HBx和对照组Fas / FasL,Bax / Bcl-2和c-myc mRNA的表达。结果:RT-PCR分析证实HBV X基因已成功转染HL-7702细胞。通过半定量RT-PCR分析,瞬时转染的HL-7702 / HBx细胞中Bax和c-myc mRNA水平明显高于对照组,稳定转染的HL-7702 / HBx细胞中FasL和c-myc mRNA水平转染显着高于对照组,而瞬时和稳定转染的HL-7702 / HBx细胞中Bcl-2 mRNA的水平显着低于对照组。结论:HBV X基因可能通过调节Fas / FasL,Bax / Bcl-2和c-myc基因的表达来促进肝细胞凋亡,并呈剂量依赖性。

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