摘要
目的:探究长链非编码RNA(LncRNA) BC002811对胃癌细胞活性、细胞间质转化及p27kip1活化的影响。方法 :收集本院原发性胃癌手术切除样本50例,采用TaqMan RNA试剂盒检测BC002811的相对表达量。将胃癌细胞株HGC-27细胞培养及转染后分为HGC-27组(无转染的HGC-27细胞)、BC002811-NC组(HGC-27细胞+BC002811 siRNA-NC)、BC002811组(HGC-27细胞+BC002811 siRNA);HGC-27组、空载体组(HGC-27细胞+p27kip1空质粒)、p27kip1组(HGC-27细胞+p27kip1质粒);HGC-27组、BC002811+空载体组(HGC-27细胞转染BC002811+p27kip1空质粒)、BC002811+p27kip1组(HGC-27细胞转染BC002811+p27kip1)。Transwell分别检测各组HGC-27细胞侵袭、转移能力,采用免疫印迹检测HGC-27细胞vimentin、E-cadherin蛋白表达,采用qRT-PCR检测HGC-27细胞BC002811、p27kip1表达。结果 :与胃癌组织比较,癌旁组织BC002811的表达显著降低。与HGC-27组比较,BC002811组HGC-27细胞侵袭、迁移能力及E-cadherin表达水平显著降低,vimentin表达水平升高,HGC-27组与BC002811-NC组差异无统计学意义。与HGC-27组比较,p27kip1组HGC-27细胞的侵袭、迁移能力及E-cadherin表达水平显著降低,vimentin表达水平升高,HGC-27组与空载体组差异无统计学意义。与HGC-27组比较,BC002811+空载体组HGC-27细胞侵袭、迁移能力及E-cadherin表达水平显著降低,vimentin表达水平升高;与BC002811+空载体组比较,BC002811+p27kip1组HGC-27细胞侵袭、迁移能力及E-cadherin表达水平显著降低,vimentin表达水平升高。Pearson相关分析结果显示,在HGC-27细胞侵袭、迁移中,BC002811与vimentin水平呈负相关性,与E-cadherin水平呈正相关;vimentin与E-cadherin水平呈负相关性。结论 :在胃癌细胞中BC002811-siRNA激活p27kip1水平、抑制胃癌细胞的迁移及侵袭,其作用机制可能与调控胃癌细胞间质转化蛋白表达相关。
Objective:To explore effect of p27kip1 activated by long noncoding RNA(LncRNA)BC002811 on the activity and interstitial transformation of gastric cancer cells.Methods:Fifty surgical resection samples of primary gastric cancer in our hospital were collected,and the relative expression of BC002811 was detected by TaqMan RNA kit.HGC-27 cells were cultured and transfected into the HGC-27 group(HGC-27 cells without transfection),BC002811-NC group(HGC-27 cells+BC002811 siRNA-NC),BC002811 group(HGC-27 cells+BC002811 siRNA),HGC-27 group,empty vector group(HGC-27 cells+p27kip1 empty plasmid),p27kip1 group(HGC-27 cells+p27kip1 plasmid);HGC-27 group,BC002811+empty vector group(HGC-27 cells transfected with BC002811+p27kip1 empty plasmid),and BC002811+p27kip1 group(HGC-27 cells transfected with BC002811+p27kip1).The invasion and metastasis of HGC-27 cells in each group detected by Transwell.The expression of vimentin and E-cadherin protein was detected by Western blotting,and the expression of BC002811 and p27kip1 was detected by qRT-PCR.Results:Compared with the gastric cancer tissue,the expression of BC002811 in the paracancerous tissue was decreased significantly.Compared with the HGC-27 group,the invasion,migration and E-cadherin expression of HGC-27 cells in BC002811 group were significantly decreased,while vimentin expression was increased.There was no difference between the HGC-27 group and BC002811-NC group.Compared with the HGC-27 group,the invasion,migration and E-cadherin expression of HGC-27 cells in the p27kip1 group were significantly decreased,while the vimentin expression was increased.There was no difference between the HGC-27 group and the empty verctor group.Compared with HGC-27 group,HGC-27 cell invasion,migration and E-cadherin expression were significantly decreased in BC002811+empty vector group,while vimentin expression was increased.Compared with BC002811+empty vector group,HGC-27 cell invasion,migration and E-cadherin in BC002811+p27kip1 group were significantly decreased,while vimentin expression was increased.Pearson correlation analysis confirmed that BC002811 was negatively correlated with vimentin expression during HGC-27 cell invasion and migration.BC002811 was positively correlated with E-cadherin expression,and vimentin expression was negatively correlated with E-cadherin expression.Conclusion:In gastric cancer,BC002811-siRNA activates the level of p27kip1,inhibits the migration and invasion of gastric cancer cells,and its mechanism of action may be related to the regulation of stromal transition protein expression of gastric cancer cells.
作者
陈娟
徐涛
薛建波
孔阳
梅倩
Chen Juan;Xu Tao;Xue Jianbo;Kong Yang;Mei Qian(Department of Gastroenterology,Daxing District People’s Hospital,Beijing 102600;Department of Gastroenterology,Chinese People’s Liberation Army General Hospital,Beijing 100039,China)
出处
《解剖学杂志》
CAS
2023年第3期190-196,F0002,共8页
Chinese Journal of Anatomy
基金
国家自然科学基金(81473268)。
