LncRNA NNT-AS1对胃癌细胞增殖、凋亡作用机制研究  

Effect of LncRNA NNT-AS1 on proliferation and apoptosis of gastric carcinoma cells and its mechanism

作　　者：帅勇锋[1] 王小军[1] 张一中[1] SHUAI Yongfeng,;WANG Xiaojun;ZHANG Yizhong.(epartment of Gastrointestinal Surgery, the Affiliated Hospital of Ningbo University School of Medicine, Ningbo 315000, China)

机构地区：[1]宁波大学医学院附属医院胃肠外科,315000

出　　处：《浙江医学》2018年第5期439-442,478,共5页Zhejiang Medical Journal

摘　　要：目的探讨LncRNA NNT-AS1对胃癌细胞增殖、凋亡的作用机制。方法采用实时荧光定量PCR法检测胃癌细胞系KATO-III、NUGC-3、AGS、N87及正常胃成纤维细胞系NSFC中LncRNA NNT-AS1的表达水平;将KATO-III细胞系分成对照组、sh-vector组及sh-NNT-AS1组,分别不感染慢病毒、感染阴性shRNA慢病毒及感染lenti-sh-NNT-AS1慢病毒。采用MTT实验检测细胞增殖情况,流式细胞术检测细胞凋亡情况,Western blot检测细胞凋亡蛋白Caspase 3、8、9的表达水平,并进行组间比较。结果胃癌细胞系KATO-III、NUGC-3、AGS、N87 LncRNA NNT-AS1的表达水平均明显高于正常胃成纤维细胞系NSFC(均P<0.01)。细胞培养12、24、48、72 h后,相比sh-vector组、对照组,sh-NNT-AS1组吸光度值(OD490值)明显降低(均P<0.05)。细胞培养7d后,sh-NNT-AS1组克隆形成数明显低于sh-vector组、对照组(均P<0.05)。sh-NNT-AS1组细胞凋亡率明显高于sh-vector组、对照组(均P<0.01)。sh-NNT-AS1组细胞凋亡蛋白Caspase3、9表达水平均高于sh-vector组(均P<0.05),而Caspase8表达水平比较差异无统计学意义(P>0.05)。结论 LncRNA NNT-AS1促进胃癌细胞增殖,抑制凋亡,机制可能与上调凋亡蛋白Caspase3、9表达有关。Objective To investigate the effect of LncRNA NNT-AS1on proliferation and apoptosis of gastric carcinoma cells and its mechanism.Methods The expression levels of LncRNA NNT-AS1in the gastric carcinoma cell lines KATO-III,NUGC-3,AGS,N87and normal gastric cell line NSFC was determined by qRt-PCR.The KATO-III cells were transfected with negative shRNA-Lenti(vector group)or Lenti-sh-NNT-AS1(sh-NNT-AS1group).The cell proliferation ability was measured by MTT assay,apoptosis was tested by flow cytometry,the expression of Caspase3,8,9protein was measured by Western blot.Results The relative express levels of LncRNA NNT-AS1in KATO-III,NUGC-3,AGS and N87cells were significantly higher than that in normal gastric NSFC cells(P<0.05).MTT assay demonstrated that the OD490value of sh-NNT-AS1group was significantly lower than control and vector group after culturing for12,24,48and72h(P<0.05).The number of clone formation cells in sh-NNT-AS1group was significantly lower than that in control group and vector group(P<0.01).The apoptosis rate of sh-NNT-AS1group was significantly higher than that in vector and control group(P<0.01).The expression level of cleaved Caspase3,9protein in sh-NNT-AS1group was significantly higher than that in vector group(P<0.05,P<0.01);while,there was no significantly difference in expression of cleaved caspase-8protein between sh-NNT-AS1and vector groups(P>0.05).Conclusion LncRNA NNT-AS1may promote proliferation and inhibit apoptosis of gastric carcinoma cells,which may is associated with up-regulation of Caspase3and9.

关 键 词：LncRNA NNT-AS1 胃癌 增殖 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]