Yao Min, Wang Li, Dong Zhizhen, Qian Qi, Shi Yun, Yu Dandan, Wang Shiye, Zheng Wenjie, Yao Dengfu
Research Center of Clinical Medicine, Affiliated Hospital of Nantong University, 20 West Temple Road, 226001, Nantong, Jiangsu Province, China.
Tumour Biol. 2014 Jun;35(6):5857-68. doi: 10.1007/s13277-014-1776-5. Epub 2014 Mar 16.
Glypican-3 (GPC-3), a membrane-associated heparan sulfate proteoglycan, plays a crucial role in cell proliferation and metastasis, particularly in hepatocellular carcinoma (HCC) progression, and perhaps is a valuable target for its gene therapy. However, its mechanism remains to be explored. In the present study, the biological behaviors of HCC cells were investigated by interfering GPC-3 gene transcription. After the cells were transfected with specific GPC-3 short hairpin RNA (shRNA), the inhibition of GPC-3 expression was 75.6 % in MHCC-97H or 73.8 % in Huh7 cells at mRNA level; the rates of proliferation and apoptosis were 53.6 and 60.5 % in MHCC-97H or 54.9 and 54.4 % in Huh7 cells, with the cell cycles arrested in the G1 phase; the incidences of cell migration, metastasis, and invasion inhibition were 80.1, 56.4, and 69.1 % in MHCC-97H or 80.9, 59.6, and 58.3 % in Huh7 cells, respectively. The cell biological behaviors were altered by silencing GPC-3 with down-regulation of β-catenin, insulin-like growth factor-II and vascular endothelial growth factor, and Gli1 up-regulation. The cell proliferation was significantly inhibited (up to 95.11 %) by shRNA plus anti-cancer drugs, suggesting that GPC-3 gene should be a potential target for promoting hepatoma cell apoptosis and inhibiting metastasis through the Wnt/β-catenin and Hh singling pathways.
磷脂酰肌醇蛋白聚糖-3（GPC-3）是一种膜相关硫酸乙酰肝素蛋白聚糖，在细胞增殖和转移中起关键作用，尤其是在肝细胞癌（HCC）进展过程中，或许是其基因治疗的一个有价值的靶点。然而，其机制仍有待探索。在本研究中，通过干扰GPC-3基因转录来研究HCC细胞的生物学行为。用特异性GPC-3短发夹RNA（shRNA）转染细胞后，在mRNA水平上，MHCC-97H细胞中GPC-3表达的抑制率为75.6%，Huh7细胞中为73.8%；MHCC-97H细胞的增殖率和凋亡率分别为53.6%和60.5%，Huh7细胞中为54.9%和54.4%，细胞周期停滞在G1期；MHCC-97H细胞的细胞迁移、转移和侵袭抑制发生率分别为80.1%、56.4%和69.1%，Huh7细胞中分别为80.9%、59.6%和58.3%。通过沉默GPC-3改变细胞生物学行为，同时β-连环蛋白、胰岛素样生长因子-II和血管内皮生长因子表达下调，Gli1表达上调。shRNA联合抗癌药物可显著抑制细胞增殖（高达95.11%），表明GPC-3基因应是通过Wnt/β-连环蛋白和Hh信号通路促进肝癌细胞凋亡和抑制转移的潜在靶点。
