Shao Zhuo, Ji Weiping, Liu Anan, Qin Ancheng, Shen Li, Li Gang, Zhou Yingqi, Hu Xiangui, Yu Enda, Jin Gang
Department of General Surgery, Changhai Hospital, Second Military Medical University, Shanghai, China (mainland).
Department of General Surgery, Changzheng Hospital, Second Military Medical University, Shanghai, China (mainland).
Med Sci Monit. 2015 Nov 5;21:3371-9. doi: 10.12659/msm.894447.
The tumor susceptibility gene 101 (TSG101) was originally identified as a tumor-suppressor gene that mediates many molecular and biological processes, such as ubiquitination, endosomal trafficking, cell survival, and virus budding, but its role in hepatocellular carcinoma (HCC) is currently unknown.
We assessed the expression of TSG101 in HCC and paracancerous tissues using qPCR. Then, we used the TSG101-specific siRNA mix to disrupt the expression of TSG101 to investigate the subsequent effect on human hepatoma-7 (Huh7) cells. Western blot was used to detect the protein expression of TSG101 and other molecules. Cell growth assay was performed using CCK8. Transwell assay was used to investigate the migration and invasion ability of Huh7 cells after transfection with of TSG101 siRNA. Flow cytometry was used to estimate the effect of TSG101 knockdown on cell cycle and apoptosis. Confocal laser scanning microscopy was used to observe the actin filaments change and the formation of autophagy.
TSG101 was over-expressed in HCC tissues. TSG101 silence was able to suppress Huh7 cell proliferation, migration, and invasion. Furthermore, silencing of TSG101 could induce cell cycle arrest at G1 phase and inhibit the expression of cyclin A and cyclin D, while up-regulating the expression of CDK2. The mechanism might be induction of autophagic cell death and inactivation of Akt and ERK1/2.
TSG101 plays an important role in the development of HCC and may be a target for molecular therapy.
肿瘤易感基因101（TSG101）最初被鉴定为一种肿瘤抑制基因，它介导许多分子和生物学过程，如泛素化、内体运输、细胞存活和病毒出芽，但其在肝细胞癌（HCC）中的作用目前尚不清楚。
我们使用qPCR评估TSG101在肝癌组织和癌旁组织中的表达。然后，我们使用TSG101特异性siRNA混合物破坏TSG101的表达，以研究其对人肝癌-7（Huh7）细胞的后续影响。使用蛋白质印迹法检测TSG101和其他分子的蛋白质表达。使用CCK8进行细胞生长测定。使用Transwell测定法研究转染TSG101 siRNA后Huh7细胞的迁移和侵袭能力。使用流式细胞术评估敲低TSG101对细胞周期和凋亡的影响。使用共聚焦激光扫描显微镜观察肌动蛋白丝的变化和自噬的形成。
TSG101在肝癌组织中过表达。TSG101沉默能够抑制Huh7细胞的增殖、迁移和侵袭。此外，沉默TSG101可诱导细胞周期停滞在G1期，抑制细胞周期蛋白A和细胞周期蛋白D的表达，同时上调CDK2的表达。其机制可能是诱导自噬性细胞死亡以及使Akt和ERK1/2失活。
TSG101在肝癌发生发展中起重要作用，可能成为分子治疗的靶点。
