含Sushi重复蛋白X连锁2基因敲除对肺腺癌A549细胞生物学特性的影响  

Effect of Sushi repeat containing protein,X-linked 2 gene knockout on the biological characteristics of lung adenocarcinoma A549 cells

作　　者：赵隽 温松 于法明[2] 姜东亮 李慎柯[1,2] ZHAO Jun;WEN Song;YU Faming;JIANG Dongliang;LI Shenke(Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Department of Respiratory,Puyang Oilfield General Hospital,Puyang 457001,Henan Province,China)

机构地区：[1]新乡医学院,河南新乡453003 [2]濮阳市油田总医院呼吸科,河南濮阳457001

出　　处：《新乡医学院学报》2022年第10期901-906,共6页Journal of Xinxiang Medical University

基　　金：2019年度河南省医学科技攻关计划联合共建项目(编号:LHGJ20191375)。

摘　　要：目的探讨含Sushi重复蛋白X连锁2(SRPX2)基因敲除对肺腺癌人类肺泡基底上皮细胞A549细胞生物学特性的影响。方法根据是否敲除SRPX2基因将对数生长期的A549细胞分为对照组和敲除组,敲除组细胞采用成簇规律间隔短回文重复序列(CRISPR)-CRISPR相关蛋白9基因编辑技术敲除SRPX2基因,对照组细胞常规传代培养。采用聚合酶链式反应法、琼脂糖凝胶电泳以及DNA测序验证是否成功敲除SRPX2基因。采用流式细胞术检测2组细胞周期,细胞计数试剂盒法检测2组细胞增殖情况,Transwell小室实验检测2组细胞迁移能力。结果琼脂糖凝胶电泳及DNA测序结果均显示敲除组细胞成功敲除68 bp SRPX2基因部分序列。培养24 h时,敲除组G_(0)/G_(1)及S期细胞所占比例显著低于对照组,G_(2)/M期细胞所占比例显著高于对照组(P<0.05);培养48 h时,敲除组S期细胞所占比例显著低于对照组,G_(2)/M期细胞所占比例显著高于对照组(P<0.05);培养48 h时,2组G_(0)/G_(1)期细胞所占比例比较差异无统计学意义(P>0.05)。培养0、24、48、72、96 h时,对照组和敲除组细胞的增殖能力随时间的延长均呈升高趋势(F=244.463、87.198,P<0.05);培养0、24、48、72、96 h时,2组细胞的增殖能力比较差异均无统计学意义(P>0.05)。培养24 h时,对照组和敲除组迁移细胞数分别为(24.200±3.899)、(6.800±1.304)个;培养48 h时,对照组和敲除组迁移细胞数分别为(92.200±6.419)、(48.800±2.280)个;培养24、48 h时,敲除组迁移细胞数显著少于对照组(t=9.464、14.247,P<0.05)。结论敲除肺腺癌A549细胞中SRPX2基因能显著抑制细胞的迁移、阻滞细胞周期,SRPX2基因可能成为肺腺癌治疗的新靶点。Objective To investigate the effect of Sushi repeat containing protein,X-linked 2(SRPX2)gene knockout on the biological characteristics of lung adenocarcinoma human alveolar basal epithelial cells A549.Methods A549 cells in logarithmic growth phase were divided into the control group and the knockout group according to whether the SRPX2 gene was knocked out.The SRPX2 gene in the cells in the knockout group was knocked out by clustered regularly interspaced short palindromic repeats(CRISPR)-CRISPR associated protein 9 gene editing technology,and the cells in the control group were routinely subcultured.Whether the SRPX2 gene was successfully knocked out was verifyed by polymerase chain reaction,agarose gel electrophoresis and DNA sequencing.The cell cycle in the two groups was detected by flow cytometry.The cell proliferation in the two groups was detected by cell counting kit.The cell migration ability in the two groups was detected by Transwell assay.Results The results of agarose gel electrophoresis and DNA sequencing showed that the cells in the knockout group were successfully knocked out the 68 bp partial sequence of SRPX2 gene.When cultured for 24 h,the ratio of cells in G_(0)/G_(1) and S phase in the knockout group were significantly lower than those in the control group,and the ratio of cells in G_(2)/M phase was significantly higher than that in the control group(P<0.05).When cultured for 48 h,the ratio of cells in S phase in the knockout group was significantly lower than that in the control group,and the ratio of cells in G_(2)/M phase was significantly higher than that in the control group(P<0.05);there was no significant difference in the ratio of cells in G_(0)/G_(1) phase between the two groups(P>0.05).When cultured for 0,24,48,72 and 96 h,the proliferation ability of cells in the control group and knockout group increased with time(F=244.463,87.198;P<0.05);at 0,24,48,72 and 96 h of culture,there was no significant difference in the proliferation ablitity of cells between the two groups(P>0.05).Whe

关 键 词：含Sushi重复蛋白X连锁2 基因敲除 肺腺癌 A549细胞 生物学特性 

分 类 号：R734.2[医药卫生—肿瘤]