敲低及过表达G6PD对肝癌细胞增殖和迁移的影响 .pdfVIP

敲低及过表达G6PD对肝癌细胞增殖和迁移的影响

冯笑;刘兆宇;胡腊;陈纪涛;曾子成;刘季芳

【摘要】目的研究在人肝癌细胞株PLC/PRF/5中,敲低及过表达G6PD对细胞株

增殖、生长及迁移能力的影响.方法包装慢病毒颗粒,选用人肝癌细胞株

PLC/PRF/5建立G6PD敲低及过表达稳转细胞株,通过PCR及Westernblot检验

过表达及敲低效果,利用建立好的细胞株进行功能实验:通过实时细胞分析系统

(RTCA)记录细胞增殖及迁移曲线,EDU实验可直观显示处于DNA复制阶段的细胞

比例,以克隆形成实验反映细胞的生长能力.结果在G6PD敲低株中,细胞的倍增时

间较对照组延长,生长速率明显下降,EDU实验中处于增殖期细胞的比例较对照组下

降43.2%,克隆形成率明显下调(P0.05),敲低组的迁移速率亦明显降低,曲线分离显

著;而在过表达株及其对照组两株细胞中,增殖、生长及迁移实验结果无明显差异.结

论在肝癌细胞中降低G6PD的表达将抑制肝癌的增殖生长,为进一步研究肝癌的发

生机制及治疗方案打下了基础.%ObjectiveToinvestigatetheeffectof

knockdownoroverexpressionofG6PDonproliferation,growthand

migrationofhumanhepatocellularcarcinomacellPLC/PRF/5.Methods

Lentivirus-mediatedknock-downoroverexpressionofG6PDwasachieved

inhumanhepatocellularcarcinomacelllinePLC/PRF/5.RT-PCRand

Westernblottingassaywereusedtodetecttheoverexpressionor

knockdownofG6PD.Cellproliferationandmi-grationcurveswere

recordedbyreal-timecellanalysissystem(RTCA),thecellproportioninthe

DNAreplicationphasecanbedirectlydisplayedwithEDUexperiment,cell

growthabilitywasdetectedbycolonyformingassay.ResultsThedoubling

timeofcellsinG6PDknockdowngroupwaslongerthanthatofthecontrol

group,andthecellgrowthratedecreasedsignificantly,theproportionof

cellsinproliferativephase(43.2%)waslowerthanthatinthecontrol

group,buttheratescolonyformationandmigrationweresignificantly

decreased(P0.05,respective-ly),andthemigrationcurvesseparated

apparently.Whilenosignificantdifferencesinproliferation,growthandmi-

grationofPLC/PRF/5cellswerefoundbetweentheover-expressedstrain

andthecontrolgroup.ConclusionThereductionofG6PDexpressionin

HCCcellsinhibitstheproliferationandgrowthofHCC,whichmaylaya

foun-dationforthefurtherstudyofthepathogenesisandtreatmentof

HCC.

【期刊名称】《实用医学杂志》

【年(卷),期】2018(034)005

【总