环状RNA-波形蛋白对结直肠癌细胞增殖和凋亡的作用及其机制  

Role and mechanism of circular RNA-vimentin in the proliferation and apoptosis of colorectal cancer cells

作　　者：姚建宁[1] 张学秀[1] 张延祯 李艳乐[1] 王春峰[1] 张连峰[1] Yao Jianning;Zhang Xuexiu;Zhang Yanzhen;Li Yanle;Wang Chunfeng;Zhang Lianfeng(Department of Gastroenterology,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院消化内科,450052

出　　处：《中华消化杂志》2019年第7期458-465,共8页Chinese Journal of Digestion

摘　　要：目的 探讨环状RNA-波形蛋白(circ-VIM)调控结直肠癌细胞增殖和凋亡的作用及其机制.方法 收集2016年12月至2017年12月,于郑州大学第一附属医院普外科行结直肠癌根治术并经术后病理检查确诊的100例结直肠癌患者的病历资料,以及其癌组织和所对应的癌旁组织(阴性对照)标本.采用实时荧光定量PCR法检测circ-VIM在结直肠癌中的表达水平.采用细胞活性检测试剂盒检测结直肠癌HCT-116和HT29细胞的增殖水平.采用膜联蛋白Ⅴ/碘化丙啶实验检测HCT-116和HT29细胞的凋亡比例.采用5,5',6,6'-四氯-1,1',3,3'-四乙基苯并咪唑羰花青碘化物(JC-1)法检测HCT-116和HT29细胞线粒体势能的变化.采用蛋白质印迹法检测蛋白激酶B/哺乳动物雷帕霉素靶蛋白(mTOR)信号通路的蛋白质表达变化.采用miRDB软件预测circ-VIM的靶miRNA.统计学分析采用t检验和卡方检验.结果 结直肠癌组织中的circ-VIM表达水平为2.387±0.536,高于所对应癌旁组织中的1.110±0.134,差异有统计学意义(t=23.096,P<0.01).circ-VIM表达水平在不同肿瘤体积、TNM分期和有无淋巴结转移患者中差异均有统计学意义(P均<0.05).lenti-circ-VIM组HCT-116和HT29细胞增殖水平分别为0.737±0.023和0.835±0.025,分别高于对照组的0.449±0.020和0.531±0.019,差异均有统计学意义(t=20.706、-15.374,P均<0.01).lenti-circ-VIM-shRNA组HCT-116和HT29细胞增殖水平分别为0.236±0.027和0.243±0.019,分别低于对照组,差异均有统计学意义(t=24.557、-23.197,P均<0.01).lenti-circ-VIM-shRNA组HCT-116和HT29细胞的凋亡比例分别为(18.00±1.82)%和(20.80±0.61)%,分别高于对照组的(6.64±2.01)%和(7.35±1.36)%,差异均有统计学意义(t=8.826、17.454,P均<0.01).lenti-circ-VIM-shRNA组HCT-116和HT29细胞中的JC-1聚合物和JC-1单体荧光强度比值分别为2.21±0.12和1.40±0.11,分别低于对照组的14.54±1.00和9.24±1.18,差异均有统计学意义(t=-19.558、-15.685,P均<0.01),说明线粒体膜�Objective To investigate the role and mechanism of circular RNA-vimentin(circ-VIM)in the proliferation and apoptosis of colorectal cancer cells.Methods From December 2016 to December 2017,at Department of General Surgery of The First Affiliated Hospital of Zhengzhou University,the clinical data of 100 patients who underwent radical resection of colorectal cancer and were confirmed by pathological examination after operation were collected.The tumor tissues and corresponding paracancerous tissues(negative control)were also collected.The expression of circ-VIM in the colorectal cancer tissues was determined by quantitative real-time polymerase chain reaction(qRT-PCR).The proliferation of HCT-116 and HT29 colorectal cancer cells was detected by cell counting kit-8 assay.The ratio of apoptosis of HCT-116 and HT29 cells was measured by annexinⅤ/propidium iodide double staining assay.The mitochondrial membrane potential of HCT-116 and HT29 cells was examined by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethyl-imidacarbocyanine iodide(JC-1)assay.The expression changes of protein kinase B and mammalian target of rapamycin were tested by Western blotting.The target miRNA of circ-VIM was predicted by miRDB software.T-test and chi-square test were performed for statistical analysis.Results The expression of circ-VIM in colorectal cancer tissues was 2.387±0.536,which was higher than that in corresponding paracancerous tissues(1.110±0.134),and the difference was statistically significant(t=23.096,P<0.01).And the expression levels of circ-VIM were significantly different in patients with different tumor size,TNM stage and lymph node metastasis(all P<0.05).The proliferation of HCT-116 cells and HT29 cells in lenti-circ-VIM group was 0.737±0.023 and 0.835±0.025,respectively,which were both higher than those in control group(0.449±0.020 and 0.531±0.019),and the differences were statistically significant(t=20.706 and-15.374,both P<0.01).The proliferation of HCT-116 cells and HT29 cells in lenti-circ-VIM-shRNA group was 0.236±0.

关 键 词：结直肠肿瘤 细胞增殖 细胞凋亡 环状RNA-VIM 

分 类 号：R73[医药卫生—肿瘤]