关键词: soluble blypican-3, glypican-3, 真核表达载体, 高转移性肝癌细胞, 生物学行为

Abstract: This work aims to construct the recombinant eukaryotic expression vector for sGPC3/GPC3（soluble glypican-3/glypican-3）gene，and to study the effects of overexpressing sGPC3/GPC3 on the biological behaviors of human highly metastatic hepatocarcinoma cells HCCLM3. The gene fragment encoding sGPC3/GPC3 was obtained by PCR and cloned into eukaryotic expression vector pcDNA3.0-GFP，then with which HCCLM3 was transfected by liposome-mediated method. The expression of GPC3 was detected by RT-PCR and Western blot. The effects of sGPC3/GPC3 on the biological behavior of high metastatic hepatocarcinoma cells were detected by CCK-8，clonal plate formation experiment，cell scratch test and flow cytometry. Results showed that the recombinant eukaryotic expression vector of sGPC3/GPC3 gene was successfully constructed by restriction enzyme digestion and sequencing. The results by RT-PCR and Western blot demonstrated that sGPC3/GPC3 was successfully overexpressed in HCCLM3. Compared with normal group and transfected control vector，the proliferation ability and clonogenic ability of HCCLM3 were significantly inhibited by the overexpression of sGPC3/GPC3（P< 0.05），the mobility significantly decreased and the cell cycle was arrested in G1 phase，thus，the rate of apoptosis increased. Moreover，the effect by overexpressing sGPC3 was significantly better than by GPC3（P < 0.05）. Conclusively，the up-regulation of sGPC3/GPC3 inhibits the growth and migration of highly metastatic hepatocarcinoma cells HCCLM3，induces the cell apoptosis and affects the redistribution of cell cycle，which plays an important role in the development of highly metastatic liver cancer.

Key words: soluble glypican-3, glypican-3, eukaryotic expression vector, highly metastatic hepatocarcinoma, biological behavior