华支睾吸虫Severin与HBV-X共同作用对肝细胞和肝癌细胞凋亡和周期影响的相关性研究
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摘要
目的研究华支睾吸虫Severin蛋白(Csseverin)与HBV-X蛋白(HBX蛋白)共同作用对肝细胞和肝癌细胞凋亡和周期的影响。方法构建表达HBX的稳转肝细胞和肝癌细胞株,经诱导凋亡后用纯化Severin蛋白孵育,检测Severin蛋白对细胞凋亡和周期影响,染色后用流式细胞术检测细胞各期凋亡比例和处于不同周期细胞比例。结果随Csseverin浓度升高,稳转HBX的肝细胞株凋亡比例显著降低,由(24.13±0.68)%降低到(10.72±0.29)%;细胞增殖期比例增加,由15.43%增加到27.72%,而空载质粒对照组无明显变化。随Csseverin浓度升高,稳转HBX的肝癌细胞株凋亡比例显著降低,由(27.75±1.21)%降低到(3.91±0.75)%;细胞增殖期比例增加,由31.96%增加到38.04%。结论 Csseverin蛋白与HBV-X共同作用有抑制肝癌细胞凋亡,促进细胞增殖的作用。
Objective To study the effect of Clonorchis sinensis severin protein(Csseverin) and HBV-X(HBX) on apoptosis and cell cycle of hepatocytes and hepatocellular carcinoma cells. Methods Apoptosis and periodic changes were detected by flow cytometry. Stable transgenic hepatocytes and liver cancer cell lines expressing HBX were constructed. After induction of apoptosis,different concentration of Csseverin was added. Flow cytometry was used to analyzed the effect of Csseverin on the percentage of apoptosis and cell cycle. Results With the increase of Csseverin concentration,the apoptotic rate of hepatocellular cell lines stably transfected with HBX decreased significantly from(24.13± 0.68)% to(10.72±0.29)%. The proportions of cell proliferation phase increased from 15.43% to 27.72%. However,there was no significant change trend in the control group with no-load plasmid. The apoptotic rate of hepatoma cell lines stably transfected with HBX decreased significantly from(27.75 ± 1.21)% to(3.91 ± 0.75)%,and the proportion of cell proliferation increased from 31.96% to 38.04%. Conclusion Csseverin protein and HBV-X could inhibit apoptosis of hepatocellular carcinoma cells and promote cell proliferation.
Objective To study the effect of Clonorchis sinensis severin protein(Csseverin) and HBV-X(HBX) on apoptosis and cell cycle of hepatocytes and hepatocellular carcinoma cells. Methods Apoptosis and periodic changes were detected by flow cytometry. Stable transgenic hepatocytes and liver cancer cell lines expressing HBX were constructed. After induction of apoptosis,different concentration of Csseverin was added. Flow cytometry was used to analyzed the effect of Csseverin on the percentage of apoptosis and cell cycle. Results With the increase of Csseverin concentration,the apoptotic rate of hepatocellular cell lines stably transfected with HBX decreased significantly from(24.13± 0.68)% to(10.72±0.29)%. The proportions of cell proliferation phase increased from 15.43% to 27.72%. However,there was no significant change trend in the control group with no-load plasmid. The apoptotic rate of hepatoma cell lines stably transfected with HBX decreased significantly from(27.75 ± 1.21)% to(3.91 ± 0.75)%,and the proportion of cell proliferation increased from 31.96% to 38.04%. Conclusion Csseverin protein and HBV-X could inhibit apoptosis of hepatocellular carcinoma cells and promote cell proliferation.
引文
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[2]Bouchard MJ,Wang LH,Schneider RJ.Calcium signaling by HBx protein in hepatitis B virus DNA replication[J].Science,2001,294(5550):2376-2378.
[3]石梦辰,黄艳,余新炳,等.华支睾吸虫severin蛋白对肝癌细胞SMMC-7402增殖、迁移及侵袭作用的初步研究[J].热带医学杂志,2016,16(2):127-130.
[4]黄艳,余新炳.食源性寄生虫病流行趋势、研究与发展方向[J].中国寄生虫学与寄生虫病杂志,2015,33(6):436-442.
[5]Shin HR,Oh JK,Masuyer E,et al.Epidemiology of cholangiocarcinoma:An update focusing on risk factors[J].Cancer Science,2010,101(3):579-585.
[6]Qian MB,Utzinger J,Keiser J,et al.Clonorchiasis[J].Lancet,2016,387(10020):800-810.
[7]石梦辰.华支睾吸虫Severin与肝/胆管癌细胞凋亡、迁移和侵袭过程的相关性研究[D].广州:中山大学,2016.
[8]Cho JE,Park W,Kim DC,et al.Down-regulation of gelsolin may play a role in the progression of inverted papilloma through an antiapoptotic mechanism[J].Am J Rhinol Allergy,2012,26(3):177-182.
[9]石梦辰.华支睾吸虫分泌排泄蛋白Severin的分子特性及其抗原发性肝癌细胞PLC凋亡作用的研究[C].2013年全国寄生虫学与热带医学学术研讨会,2013.
[10]郑丹华,王小众.乙型肝炎病毒X蛋白的研究进展[J].中国病原生物学杂志,2018,13(10):1158-1161.
[11]池雪静,黄艳,余新炳.HBV-X蛋白与肝细胞癌关系的研究进展[J].热带医学杂志,2017,17(12):1690-1693.
[12]王玉凤,何松.HBV-X突变与TGF-β1致肝细胞肝癌相关性分析[J].重庆医科大学学报,2018,43(5):646-650.2019-03-29