Department of Hepatobiliary Surgery, Union Hospital, Fujian Medical University, Fuzhou 350001, China.
Front Biosci (Landmark Ed). 2012 Jan 1;17(4):1589-98. doi: 10.2741/4005.
We recently determined that synuclein-gamma (SNCG) is highly expressed in human gallbladder cancer (GBC), and its abnormal expression is associated with tumor aggressiveness. To investigate the effects of SNCG gene silencing on the tumorigenic profiles of the GBC cell line, NOZ, short-hairpin RNA (shRNA) interference was employed. Specifically, the SNCG transcript was targeted by SNCG-shRNA lentiviral particles designed to silence SNCG gene expression. Following selection of NOZ cells stably expressing SNCG-shRNA, SNCG expression was examined by western blot and semi-quantitative RT-PCR analyses. Phenotypic hallmarks of gallbladder carcinogenesis were assayed by CCK-8, soft agar (colony formation), modified Boyden-Chamber (invasion), and flow cytometry (cell-cycle and apoptosis) assays. Our results showed that SNCG gene silencing in NOZ cells inhibited cell growth, colony formation, and invasion. In addition, it directly increased the effectiveness of paclitaxel in inducing G2/M cell-cycle arrest and cell apoptosis. Data from our in vivo study showed a decrease in tumor growth and weight in mice injected with SNCG-silenced NOZ cells. Together, these findings suggest that SNCG plays an important role in the progression of GBC.
我们最近发现，γ-突触核蛋白（SNCG）在人胆囊癌（GBC）中高度表达，其异常表达与肿瘤侵袭性有关。为了研究 SNCG 基因沉默对 GBC 细胞系 NOZ 的致瘤特性的影响，我们采用短发夹 RNA（shRNA）干扰进行了研究。具体来说，设计了靶向 SNCG 转录本的 SNCG-shRNA 慢病毒颗粒来沉默 SNCG 基因表达。在稳定表达 SNCG-shRNA 的 NOZ 细胞中进行选择后，通过 Western blot 和半定量 RT-PCR 分析检测 SNCG 表达。通过 CCK-8、软琼脂（集落形成）、改良 Boyden-Chamber（侵袭）和流式细胞术（细胞周期和凋亡）检测胆囊癌发生的表型特征。我们的结果表明，NOZ 细胞中 SNCG 基因沉默抑制细胞生长、集落形成和侵袭。此外，它直接增加了紫杉醇诱导 G2/M 细胞周期阻滞和细胞凋亡的效果。体内研究数据表明，注射 SNCG 沉默的 NOZ 细胞的小鼠肿瘤生长和重量减少。综上所述，这些发现表明 SNCG 在 GBC 的进展中发挥重要作用。
