摘要
目的分析微小RNA(miRNA,miR)-95与胃癌细胞增殖、凋亡的关系,并探讨其作用机制。方法实时定量聚合酶链反应(Real-time PCR)法检测70例胃癌手术标本的肿瘤组织及癌旁黏膜组织中miR-95的表达;记录患者的临床病理特征,分析miR-95与患者这些临床病理特征之间的关系。应用miR-95抑制物(anti-miR-95)转染人胃癌细胞株BGC823,检测转染前后细胞活性、细胞周期、凋亡率;同时应用Real-time PCR及Western blot检测增殖及凋亡相关基因增殖细胞核抗原(PCNA)、细胞周期蛋白(Cyclin)D1、p21、生存素(Survivin)、Livin、Bad的表达。结果胃癌组织中miR-95水平明显高于癌旁黏膜组织(t=-3.399,P<0.05)。miR-95表达与肿瘤的分化程度及淋巴结转移情况有关:分化差的肿瘤组织miR-95水平明显高于分化好者,存在淋巴结转移的肿瘤组织miR-95水平明显高于无淋巴结转移者(t=-3.021,P<0.05;t=3.348,P<0.05)。胃癌细胞株AGS、SGC7901、MKN45、BGC823及胃上皮细胞株GES-1中,miR-95在胃癌细胞株表达均高于GES-1,在BGC823细胞表达最高(F=5.998,P<0.05)。anti-miR-95转染BGC823后细胞的活性明显降低,细胞G0/G1期比例升高(F=39.758,P<0.01),而G2/M(F=10.349,P<0.05)、S期比例均降低(F=15.118,P<0.01);anti-miR-95转染后凋亡率明显增高(F=25.182,P<0.01);转染后增殖、凋亡相关基因PCNA、Cyclin D1、Survivin的mRNA和蛋白表达明显降低,而p21、Bad的mRNA和蛋白表达明显增高(P<0.05)。结论胃癌组织中miR-95的表达增高,与肿瘤分化及淋巴结转移有关,miR-95可能通过调节一些增殖、凋亡基因表达而参与胃癌进展。
Objective To investigate the role of microRNA(miRNA,miR)-95 in the proliferation and apoptosis of gastric cancer cells and the possible mechanism.Methods Real-time quantitative polymerase chain reaction(Real-time PCR)was used to detect the expression of miR-95 in tumor tissues and gastric mucosal tissues from 70 patients with gastric cancer.The patient’s clinicopathological features were recorded and the relationship between miR-95 and these clinicopathologic features was analyzed.The human gastric cancer cell line BGC823 was transfected with anti-miR-95 and the cell viability,cell cycle and apoptosis rate were examined before and after transfection.Simultaneously,Real-time PCR and Western blotting were used to detect the expression of proliferation-and apoptosis-related genes proliferating cell nuclear antigen(PCNA),Cyclin D1,p21,Survivin,Livin and Bad.Results The miR-95 level in gastric cancer was significantly higher than that in gastric mucosal tissues(t=-3.399,P<0.05).Its expression was related to the degree of tumor differentiation and lymph node metastasis.The level of miR-95 in poorly differentiated tumors and in tumors with lymph node metastasis was significantly higher than that in well-differentiated tumors and without lymph node metastasis(t=-3.021,P<0.05;t=3.348,P<0.05).In gastric cancer cell lines AGS,SGC7901,MKN45 and BGC823,and gastric epithelial cell line GES-1,the miR-95 expression was higher in gastric cancer cell lines than GES-1,and highest in BGC823 cells(F=5.998,P<0.05).After transfection of anti-miR-95,the activity of the cells significantly decreased,and the ratio of G0/G1 phase increased(F=39.758,P<0.01),but the ratio of G2/M(F=10.349,P<0.05)and S phases decreased(F=15.118,P<0.01);apoptosis rate increased significantly after anti-miR-95 transfection(F=25.182,P<0.01).The mRNA and protein expression levels of PCNA,Cyclin D1 and Survivin significantly decreased,and those of p21 and Bad increased(P<0.05).Conclusion The up-regulated expression of miR-95 in gastric cancer is associated with tumor differentiation and lymph node metastasis,which may be involved in the progression of gastric cancer by regulating the expression of some proliferative and apoptotic genes.
作者
温转
檀碧波
李勇
赵群
范立侨
王冬
尔丽绵
Wen Zhuan;Tan Bibo;Li Yong;Zhao Qun;Fan Liqiao;Wang Dong;Er Limian(Quality Control Office,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China;The Third Department of Surgery,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China;Department of Endoscopy,the Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China)
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2019年第4期748-751,共4页
Chinese Journal of Experimental Surgery
基金
河北省医学科学研究重点课题计划项目(20170145).
关键词
胃癌
微小RNA-95
增殖
凋亡
临床病理特征
Gastric cancer
MicroRNA-95
Proliferation
Apoptosis
Clinicopathological features
