军事医学
軍事醫學
군사의학
BULLETIN OF THE ACADEMY OF MILITARY MEDICAL SCIENCES
2015年
8期
593-596,601
,共5页
ALKBH5%肝癌%细胞增殖
ALKBH5%肝癌%細胞增殖
ALKBH5%간암%세포증식
ALKBH5%liver cancer%cell proliferation
目的：探讨AlkB同源蛋白5（AlkB homologue 5，ALKBH5）对正常肝细胞系L－02和肝癌细胞系HepG2的增殖、周期、凋亡的影响。方法通过慢病毒稳定转染ALKBH5基因的重组载体（pEGFP－C1b－ALKBH5）至肝癌细胞系HepG2和肝细胞系L－02中，Western印迹鉴定绿色荧光蛋白（ GFP－ALKBH5）的稳定表达；实验分GFP－ALKBH5慢病毒组和GFP慢病毒组；细胞增殖实验（cell counting kit－8，CCK－8）检测两组细胞的增殖能力；流式细胞术检测两组细胞周期和细胞凋亡的情况；平板克隆形成实验检测两组细胞的生长能力。结果与GFP对照组相比，过表达GFP－ALKBH5可显著抑制HepG2和L－02细胞的增殖，引起HepG2和L－02细胞的周期阻滞，但不影响细胞的凋亡。结论 ALKBH5可显著抑制肝癌细胞HepG2和肝细胞L－02的增殖能力，扮演了抑癌基因的角色，可能在肝细胞癌的发生和发展中起重要作用。
目的：探討AlkB同源蛋白5（AlkB homologue 5，ALKBH5）對正常肝細胞繫L－02和肝癌細胞繫HepG2的增殖、週期、凋亡的影響。方法通過慢病毒穩定轉染ALKBH5基因的重組載體（pEGFP－C1b－ALKBH5）至肝癌細胞繫HepG2和肝細胞繫L－02中，Western印跡鑒定綠色熒光蛋白（ GFP－ALKBH5）的穩定錶達；實驗分GFP－ALKBH5慢病毒組和GFP慢病毒組；細胞增殖實驗（cell counting kit－8，CCK－8）檢測兩組細胞的增殖能力；流式細胞術檢測兩組細胞週期和細胞凋亡的情況；平闆剋隆形成實驗檢測兩組細胞的生長能力。結果與GFP對照組相比，過錶達GFP－ALKBH5可顯著抑製HepG2和L－02細胞的增殖，引起HepG2和L－02細胞的週期阻滯，但不影響細胞的凋亡。結論 ALKBH5可顯著抑製肝癌細胞HepG2和肝細胞L－02的增殖能力，扮縯瞭抑癌基因的角色，可能在肝細胞癌的髮生和髮展中起重要作用。
목적：탐토AlkB동원단백5（AlkB homologue 5，ALKBH5）대정상간세포계L－02화간암세포계HepG2적증식、주기、조망적영향。방법통과만병독은정전염ALKBH5기인적중조재체（pEGFP－C1b－ALKBH5）지간암세포계HepG2화간세포계L－02중，Western인적감정록색형광단백（ GFP－ALKBH5）적은정표체；실험분GFP－ALKBH5만병독조화GFP만병독조；세포증식실험（cell counting kit－8，CCK－8）검측량조세포적증식능력；류식세포술검측량조세포주기화세포조망적정황；평판극륭형성실험검측량조세포적생장능력。결과여GFP대조조상비，과표체GFP－ALKBH5가현저억제HepG2화L－02세포적증식，인기HepG2화L－02세포적주기조체，단불영향세포적조망。결론 ALKBH5가현저억제간암세포HepG2화간세포L－02적증식능력，분연료억암기인적각색，가능재간세포암적발생화발전중기중요작용。
Objective To investigate the effect of AlkB homologue 5 ( ALKBH5 ) on proliferation, cell cycle, and apoptosis of HepG2 and L-02 cells.Methods Recombinant plasmid vector containing the CDS region of ALKBH5 (pEGFP-C1b-ALKBH5) was stably transfected into HepG2 and L-02 cells.Western blotting was used to detect the expression of green fluonescence protein ( GFP )-ALKBH5.There were two groups in our experiment: GFP-ALKBH5 lentivirus group and GFP lentivirus group.Characteristics, such as proliferation, cell cycle and apoptosis of HepG2 and L-02,were detected through Cell Counting Kit-8 (CCK-8) assay, flow cytometry and clone formation, respectively.Results The result of Western blotting revealed that ALKBH5 was efficiently up-regulated at protein levels.Despite apoptosis, phenotypic analysis revealed that the proliferation and cell phases were significantly inhibited in ALKBH5 overexpressed stable cell strains compared with the control cells (both P<0.05).Conclusion ALKBH5 can restrain fetal liver cell (L-02) and hepatocellular carcinoma cell (HepG2) from proliferating.Taken together, our results strongly suggest that ALKBH5 can play a key role in the generation and progression in HCC as a tumor suppressor.
