下调GnTⅣa表达对肝癌Hca-F细胞增殖及侵袭的影响  

Effects of down-regulation of lentivirus-mediated GnT Ⅳ a on the proliferation and invasion of mouse hepatoma Hca-F cells

作　　者：樊建楚[1] 汪淑晶[1] 于生金[1,2] 何静娜 郑伟龙[1] 张嘉宁[1] 

机构地区：[1]大连医科大学糖生物学研究所,生物化学与分子生物学教研室,辽宁大连116044 [2]辽东学院医学院基础医学部,辽宁丹东118003

出　　处：《中国肿瘤生物治疗杂志》2013年第6期673-678,共6页Chinese Journal of Cancer Biotherapy

基　　金：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)~~

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-acetylglucosaminyltransferaseⅣa,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-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.

关 键 词：N-乙酰氨基葡萄糖转移酶Ⅳa 肝癌 增殖 Hca-F细胞 侵袭 慢病毒 RNAI 

分 类 号：R735.7[医药卫生—肿瘤] R730.54[医药卫生—临床医学]