Wang Peng, Wang Zhi-wei, Qian Hai-xin, Guo Qing-song
Department of Hepatobiliary Surgery,Affiliated Hospital of Nantong University, Nantong 226000, China.
Email:
Zhonghua Yi Xue Za Zhi. 2013 Nov 26;93(44):3556-8.
To investigate the role of autophagy in the injury of HepG-2 cells induced by hepatitis B virus x protein (HBx).
After HBx transfection, the cells were used to detect the formation of autophagosomes and observed by transmission electron microscopy, monodansylcadaverine (MDC) staining autophagic vacuole (AV), immunofluorescent ce staining microtubule-associated protein light chain 3 ( MAP1-LC3 ) protein, and Western blotting examining the ratio of LC3-II/LC3-I (gray level: 0.760 ± 0.078 vs 0.520 ± 0.086, P < 0.05), beclin 1 (gray level: 0.875 ± 0.093 vs 0.220 ± 0.087, P < 0.05)and lysosome associated membrane protein 2a ( lamp2a ) protein (gray level: 0.320 ± 0.061 vs 0.120 ± 0.064, P < 0.05) levels.
(1) HBx transfected upregulated the expression of LC3-II, LC3-I, beclin 1 and lamp2a protein. (2) HBx transfected brought about an increase in the formation of autophagosomes and autolysosomes.
HBx activates the autophagic lysosome pathway in HepG-2 cells through the LC3/beclin1 pathway.
探讨自噬在乙型肝炎病毒X蛋白（HBx）诱导的HepG-2细胞损伤中的作用。
转染HBx后，检测细胞自噬体的形成，通过透射电子显微镜观察，用单丹磺酰尸胺（MDC）染色自噬泡（AV），免疫荧光细胞化学法检测微管相关蛋白轻链3（MAP1-LC3）蛋白，并采用蛋白质免疫印迹法检测LC3-II/LC3-I的比值（灰度值：0.760±0.078比0.520±0.086，P<0.05）、beclin 1（灰度值：0.875±0.093比0.220±0.087，P<0.05）和溶酶体相关膜蛋白2a（lamp2a）蛋白（灰度值：0.320±0.061比0.120±0.064，P<0.05）水平。
（1）转染HBx后上调了LC3-II、LC3-I、beclin 1和lamp2a蛋白的表达。（2）转染HBx后自噬体和自溶酶体的形成增加。
HBx通过LC3/beclin1途径激活HepG-2细胞中的自噬溶酶体途径。
