目的：探讨慢病毒介导的N-乙酰氨基葡萄糖转移酶Ⅳa （N-acetylglucosaminyltransferase Ⅳa，GnTⅣa）表达下调对小鼠肝癌Hca-F细胞体外增殖及侵袭能力的影响。方法：针对GnTⅣa的编码基因Mgat4a，构建含shRNA干扰序列的慢病毒表达载体pll3.7/Mgat4a-shRNA并包装慢病毒颗粒，感染Hca-F细胞。RT-PCR 及流式细胞术分别检测Mgat4a-shRNA重组慢病毒感染对Hca-F细胞Mgat4a mRNA及GnTⅣa表达的影响，CCK-8法和Transwell侵袭实验分别检测Mgat4a-shRNA重组慢病毒感染在体外对Hca-F细胞增殖和侵袭能力的影响。结果：慢病毒能够高效感染小鼠肝癌细胞Hca-F，Mgat4a-shRNA组感染效率达72%。与Hca-F细胞和Mock-shRNA组相比，Mgat4a-shRNA组细胞内Mgat4amRNA\[（0.53±0.2） vs（2.97±02）、（2.90±0.1）；均P<0.05\]和GnTⅣa\[（47.80±2.25）vs（394.61±5.27）、（378.61±4.38）；均P<0.05\]表达显著降低；增殖能力\[细胞增殖抑制率：（46.9±0.02）%vs （0.82±0.03）%、 0，P<0.01\]和体外侵袭能力\[穿过滤膜的细胞数：（18±3）vs （42±4）、（38±3）个，均P<0.05\]显著下降。结论：慢病毒介导shRNA下调GnTⅣa的表达能够抑制小鼠肝癌细胞 Hca-F的增殖及体外侵袭。

Objective:To investigate the effect of down-regulation of lentivirus-mediated N-acetylglucosaminyltransferase Ⅳa (GnTⅣa) on the proliferation and invasion of mouse hepatoma Hca-F cells in vitro. Methods: Expression plasmid pll3.7/Mgat4a-shRNA with interference shRNA sequences gene target Mgat4agene, were encoding GnTⅣa was constructed. The RNAi lentiviral particles were packaged to infect Hca-F cells. The effect of Mgat4a-shRNA infection on the Mgat4a mRNA and GnTⅣa expression of Hca-F cells was detected by RT-PCR and flowcytometry, respectively. The effect of recombinant lentiviral infection on the proliferation and invasion capability of Hca-F cells was determined by CCK-8 and transwell assay in vitro, respectively. Results: The mouse hepotoma Hca-F cells could be infected by the lentivirus with a high efficiency, and the infection efficiency of Mgat4a-shRNA group reached 72%. Compared with the Hca-F cells and Mock-shRNA group, the Mgat4a mRNA (\[0.53±0.2\] vs \[2.97±0.2\], \[2.90±0.1\]; P<0.05) and GnTⅣa (\[47.80±2.25\] vs \[39461±5.27), \[378.61±4.38\]) expression of the Mgat4a-shRNA group was significantly decreased, and the cell proliferation (inhibitory rate \[46.9±0.02\]% vs \[0.82±0.03\]%, 0; all P<0.01) and invasion (invasive cell numbers: \[18±3\] vs \[42±4\], \[38±3\], all P<0.05) capability were inhibited significantly. Conclusion: Down-regulation of lentivirus-mediated GnTⅣa expression inhibits the proliferation and invasion of mouse hepatoma Hca-F cells in vitro.

国家重点基础研究发展计划（973计划）资助项目（No. 2012CB822103）；国家自然科学基金资助项目（No. 31000372，31170774）