摘要
目的探讨microRNA-133(miR-133)在胃癌组织及细胞系中的表达情况,并探讨其对胃癌细胞凋亡和侵袭的影响。方法采用实时定量PCR(q PCR)检测64例胃癌组织及对应癌旁组织中的miR-133水平,分析miR-133表达与临床病理参数(性别、年龄、肿瘤位置、临床分期、分化程度、浸润深度及淋巴结转移)的关系,并检测其在胃癌细胞株AGS、SGC-7901、MKN-1、MKN-45、MGC-803和BGC-823及正常胃黏膜细胞GES-1细胞的表达情况,同时选取miR-133水平最低的胃癌细胞并转染过表达miR-133的真核重组质粒p CDNA3.1+miR-133,转染后分别采用流式细胞仪PI/Annexin V双染法及Transwell法检测miR-133对细胞凋亡和侵袭能力的影响。结果胃癌组织的miR-133相对表达量为0.347±0.024,低于癌旁组织(P<0.05),且其表达与临床分期、分化程度、浸润深度及淋巴结转移有关均有关(P<0.05);与GES-1细胞相比(其miR-133表达水平设为1.00),胃癌细胞的miR-133水平均较低(P<0.05),且MKN-45的表达水平最低;与对照组和空转染组相比,过表达组转染48、96 h后的细胞凋亡水平均升高,且穿膜细胞数均降低(P<0.05)。结论 miR-133在胃癌组织和细胞中均为低表达,上调miR-133水平可抑制胃癌细胞的侵袭并诱导凋亡。
Objective To investigate the expression of microRNA-133 （miR-133） in gastric cancer tissues and cell lines as well as its effect on apoptosis and invasion of gastric cancer cells. Methods The real-time quantitative PCR （qPCR） was used to de- tect the miR-133 level in 64 eases of gastric cancer tissues and corresponding adjacent normal tissues. Then, we analyzed the relation- ship between miR-133 expression and clinical pathological parameters （age, gender, tumor location, clinical stage, differentiation de- gree, invasion depth and lymph node metastasis）. The expression of miR-133 was detected in gastric cancer cell line AGS, SGC-7901, MKN-1, MKN-45, MGC-803, BGC-823 and GES-1, and the normal gastric mucosa cell line, GES-1, was used as the control. The gastric cancer cell with the lowest level of miR-133 was chosen for the transfection with the recombinant plasmid pCDNA3, 1＋miR-133. PI/Annexin V double staining and Transwell assay were used to detect the effect of miR-133 on apoptosis and invasion of cells. Results The relative expression of miR-133 in gastric carcinoma tissues was 0. 347±0. 024, lower than that of the adjacent tissues （P〈 0.05 ）. Its expression was related to clinical stage, differentiation, depth of invasion and lymph node metastasis （P〈0. 05）. Compared with the GES-1 cells, the miR-133 levels of gastric cancer cells were lower （P〈0.05） , and the expression level of MKN-45 was low- est. Compared with other two groups at 48 and 96 h after transfection, the apoptosis level was higher but the number of the cells was de- creased in the overexpression group （P〈0.05）. Conclusion miR-133 was lowly expressed in gastric cancer tissues and cells, and up- regulation of miR-133 can inhibit the invasion and apoptosis of gastric cancer cells.
出处
《临床肿瘤学杂志》
CAS
2015年第7期588-592,共5页
Chinese Clinical Oncology
基金
国家自然科学基金资助项目(81260349)
海南省自然科学基金项目(309063)
海南省应用技术研发与示范推广专项项目(ZDXM2015080)
关键词
胃癌
miR-133
增殖
侵袭
凋亡
Gastric cancer
miR-133
Proliferation
Invasion
Apoptosis
