Kuo Chan-Yen, Wang Jing-Chyi, Hsu Shih-Lan, Hwang Guang-Yuh
Department of Life Science, Tunghai University, Taichung, Taiwan.
Intervirology. 2008;51(4):253-60. doi: 10.1159/000158522. Epub 2008 Sep 30.
This study aimed to determine the effects of HBx on the inhibition of tumorigenesis in nude mice injected with CCL13-HBx cells. Therefore, the characteristics of the induced tumors and the phenomenon of apoptosis were assessed.
The induced tumors were identified using the specific marker of hepatocellular carcinoma (HCC), anti-alpha-fetoprotein (AFP), and their characteristics were pathologically examined. Apoptosis was detected by DNA fragmentation, and the expression of the proapoptotic proteins p53, Bax, Bad, caspase-3, and caspase-8 and the anti-apoptotic protein Bcl-2 was detected by Western blotting. To identify possible molecules involved in the inhibition of tumorigenesis, extracts of the induced tumors were separated by 2D-PAGE, and the proteins were identified by MS.
The tumors of the nude mice injected with CCL13 and CCL13-HBx cells were identified as HCCs. Moreover, HBx was found to suppress tumor growth via apoptosis in the nude mice injected with CCL13-HBx cells. The MS findings revealed that phosphorylated myosin light chain was a candidate molecule involved in the inhibition of tumorigenesis.
HBx suppressed tumorigenesis in the nude mice injected with CCL13-HBx cells, which proved to be a good animal model for the in vivo study of the effects of HBx on tumorigenesis.
本研究旨在确定HBx对注射CCL13 - HBx细胞的裸鼠肿瘤发生抑制作用的影响。因此，对诱导肿瘤的特征及凋亡现象进行了评估。
使用肝细胞癌（HCC）的特异性标志物抗甲胎蛋白（AFP）鉴定诱导肿瘤，并对其特征进行病理检查。通过DNA片段化检测凋亡，通过蛋白质印迹法检测促凋亡蛋白p53、Bax、Bad、半胱天冬酶 - 3和半胱天冬酶 - 8以及抗凋亡蛋白Bcl - 2的表达。为了鉴定参与肿瘤发生抑制的可能分子，通过二维聚丙烯酰胺凝胶电泳（2D - PAGE）分离诱导肿瘤的提取物，并通过质谱（MS）鉴定蛋白质。
注射CCL13和CCL13 - HBx细胞的裸鼠肿瘤被鉴定为肝癌。此外，发现HBx通过注射CCL13 - HBx细胞的裸鼠中的凋亡抑制肿瘤生长。质谱结果显示磷酸化肌球蛋白轻链是参与肿瘤发生抑制的候选分子。
HBx抑制了注射CCL13 - HBx细胞的裸鼠中的肿瘤发生，这被证明是用于体内研究HBx对肿瘤发生影响的良好动物模型。
