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作 者:张智[1] 郭鹏[2] 孙兴[1] 肖艺[1] 徐静[3] 彭涛[3] 赵国良[3] 吴曙粤[4] 黄海[1]
机构地区:[1]南宁市第一人民医院肝胆胰腺外科,南宁530022 [2]广西医科大学附属口腔医院口腔外科,南宁530021 [3]广西医科大学第一附属医院肝胆外科,南宁530021 [4]南宁市第一人民医院儿科,南宁530022
出 处:《中国癌症防治杂志》2017年第4期306-310,共5页CHINESE JOURNAL OF ONCOLOGY PREVENTION AND TREATMENT
基 金:广西医科大学青年科学基金资助项目(GXMUYSF201542);南宁市科学研究与技术开发计划资助项目(20163130)
摘 要:目的探讨过表达乙型肝炎病毒X基因(hepatitis B virus X gene,HBx)上调热休克转录因子1(heat shock transcription factor 1,HSF1)对肝癌Hep G2细胞生长的影响。方法通过pc DNA3.1载体与HBx基因连接构建pc DNA3.1-HBx过表达质粒,同时构建空载体pc DNA3.1对照质粒,采用脂质体介导转染肝癌Hep G2细胞,获得Mock/Hep G2细胞(对照组)和HBx/Hep G2细胞(实验组)。采用Western blot技术检测细胞内HSF1和HBx蛋白表达水平,MTT实验、平板克隆形成实验、划痕实验和Transwell实验观察细胞生长情况。结果与转染空载体pc DNA3.1的Mock/Hep G2细胞相比,转染过表达载体pc DNA3.1-HBx的HBx/Hep G2细胞中HBx和HSF1蛋白表达明显增加;细胞平板克隆形成[(46.13±1.25%vs 86.43±1.01%)]、侵袭能力[(22.47±2.05)%vs(51.01±1.75)%]和迁移能力[(24.18±0.98)%vs(59.03±0.83)%]明显增强,差异有统计学意义(P<0.05)。结论 HBx基因可上调肝癌Hep G2细胞HSF1和HBx蛋白的表达,促进细胞增殖、侵袭和迁移,导致细胞恶性生长。Objective To explore the effects of the hepatitis B virus X gene(HBx) on expression of heat shock transcription factor 1 (HSF1) and growth of HepG2 hepatocellular carcinoma cells. Methods HepG2 cells were transfected with empty pcDNA3.1 vector or pcDNA3.1 encoding HBx using Lipofectamine 2000. After 24 h,the two groups of transfected cells were compared in terms of HSF1 and HBx protein expression (Western blot),MTT assay,plate clone formation,wound healing,and Transwell assay. Results HBx and HSF1 proteins were expressed at significantly higher levels in the cells transfected with pcDNA3.1-HBx than in cells transfected with empty vector. Cells transfected with the HBx gene showed significantly greater colony formation(86.43±1.01% vs 46.13±1.25%, P〈0.05 ), invasion (51.01 ±1.75% vs 22.47±2.05% ,P〈0.05 ) and migration (59.03±0.83% vs 24.18 ±0.98% ,P〈0.05 ). Conclusion Transfecting the HBx gene into HepG2 cells up-regulates expression of HBx and HSF1 ,which promotes the growth of hepatoma cells.
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