中文摘要：

目的：探讨慢病毒介导的N-乙酰氨基葡萄糖转移酶Ⅳa（N-acetylglucosaminyltran8feraseIVa，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p113．7／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Mgat4amRNA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Mgat4amRNA[（0．53±0．2）VS（2．97±0．2）、（2．90±0．1）；均P〈0．05]和GnTIVa[（47．80±2．25）口s（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）％w（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）摊（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。

英文摘要：

Objective：To investigate the effect of down-regulation of lentivirus-mediated N-acetylglucosaminyltrans ferase 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 GnT[Va 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 （ E 0.53 ± 0.2 ] vs [ 2.97± 0.2 ], [ 2.90 ± 0.1 ] ; P 〈 0.05 ） and GnT1V a （ [47.80 ±2.25] vs [394. 61 ±5.27）, [378.61 ±4.38] ） expression of the Mgat4a-shRNA group was significantly de creased, and the cell proliferation （inhibitory rate E46.9 ± 0.02 ] % vs [O. 82 ± 0.03 ] %, 0; all P 〈 0.01 ） and invasion（ invasive cell numbers ： [ 18 ＋ 31 vs [42 ± 4 ], [ 38± 3 ], all P 〈 0.05 ） capability were inhibited significantly. Conclu- sion： Down-regulation of lentivirus-mediated GnTIV a expression inhibits the proliferation and invasion of mouse hepatoma Hca-F cells in vitro.