摘要
目的探讨下调长链非编码RNA(lncRNA)HOXA-AS3靶向促进微小RNA-29b(miR-29b)对结肠癌细胞LOVO凋亡和奥沙利铂敏感性的影响。方法实时聚合酶链反应测定结肠癌细胞LOVO、SW1116、HCT116、SW620和正常结肠上皮细胞NCM460中HOXA-AS3表达变化。结肠癌细胞LOVO分成control组、si-NC组、si-HOXA-AS3组、L-OHP+si-NC组、L-OHP+si-HOXA-AS3组、si-HOXA-AS3+Anti-miR-NC组、si-HOXA-AS3+Anti-miR-29b组、L-OHP+si-HOXA-AS3+Anti-miR-NC组和L-OHP+si-HOXA-AS3+Anti-miR-29b组。采用MTT实验、流式细胞术和Western blot分别分析细胞增殖、凋亡以及细胞中Bax、Bcl-2蛋白表达情况。生物学信息学软件预测HOXA-AS3的靶miRNA,荧光素酶活性测定试剂盒验证HOXA-AS3和miR-29b的靶向关系。结果与正常结肠上皮细胞NCM460相比,结肠癌细胞LOVO、SW1116、HCT116、SW620中HOXA-AS3表达水平均升高,差异有统计学意义(P<0.05)。结肠癌细胞LOVO中HOXA-AS3表达水平均高于结肠癌细胞SW1116、HCT116、SW620,差异有统计学意义(P<0.05)。与control组、si-NC组比较,si-HOXA-AS3组结肠癌细胞LOVO增殖活性降低,凋亡率升高,细胞中Bax蛋白表达水平升高,Bcl-2蛋白表达减少,差异有统计学意义(P<0.05)。与si-HOXA-AS3组、L-OHP+si-NC组比较,L-OHP+si-HOXA-AS3组结肠癌细胞LOVO增殖活性降低,凋亡率升高,细胞中Bax蛋白表达水平升高,Bcl-2蛋白表达减少,差异有统计学意义(P<0.05)。与si-HOXA-AS3+Anti-miR-NC组比较,si-HOXA-AS3+Anti-miR-29b组结肠癌细胞LOVO增殖活性升高,细胞凋亡率降低,细胞中Bax蛋白表达减少,Bcl-2蛋白表达增多,差异有统计学意义(P<0.05)。与L-OHP+si-HOXA-AS3+Anti-miR-NC组比较,L-OHP+si-HOXA-AS3+Anti-miR-29b组结肠癌细胞LOVO增殖活性升高,细胞凋亡率降低,细胞中Bax蛋白表达减少,Bcl-2蛋白表达增多,差异有统计学意义(P<0.05)。下调HOXA-AS3靶向促进miR-29b表达。结论下调HOXA-AS3可通过靶向促进miR-29b诱导结肠癌细胞LOVO凋亡并提高奥沙利�
Objective To investigate effect of down-regulating lncRNA HOXA-AS3 targeted promotion of miR-29 b on the apoptosis of colon cancer cells LOVO and the sensitivity of oxaliplatin. Methods Real-time polymerase chain reaction was used to detect the changes of HOXA-AS3 expression in colon cancer cells LOVO, SW1116, HCT116, SW620 and normal colon epithelial cells NCM460. Colon cancer cells LOVO were divided into control group, si-NC group, si-HOXA-AS3 group, L-OHP+si-NC group, L-OHP+si-HOXA-AS3 group, si-HOXA-AS3+Anti-miR-NC group, si-HOXA-AS3+Anti-miR-29 b group, L-OHP+si-HOXA-AS3+Anti-miR-NC group and L-OHP+si-HOXA-AS3+Anti-miR-29 b group. MTT assay, flow cytometry and Western blot were used to analyze cell proliferation, apoptosis and protein expression of Bax and Bcl-2, respectively. The bioinformatics software predicted the target miRNA of HOXA-AS3, and the luciferase activity assay kit was used to verify the targeting relationship between HOXA-AS3 and miR-29 b. Results Compared with normal colonic epithelial cells NCM460,the expression of HOXA-AS3 in colon cancer cells LOVO,SW1116,HCT116 and SW620 were significantly higher( P < 0. 05). The expression level of HOXA-AS3 in colon cancer cell LOVO was significantly higher than colon cancer cell SW1116,HCT116 and SW620( P < 0. 05). Compared with the control and si-NC groups,the si-HOXA-AS3 colon cancer cell LOVO proliferation activity decreased,the apoptosis rate increased,the Bax protein expression level in the cells increased,and the Bcl-2 protein expression decreased( P < 0. 05). Compared with the si-HOXA-AS3 and L-OHP + si-NC groups,the colon cancer cell LOVO proliferation activity in the L-OHP + si-HOXA-AS3 group decreased,the apoptosis rate increased,and the expression level of Bax protein in the cells increased,the expression of Bcl-2 protein decreased( P < 0. 05). Compared with the si-HOXA-AS3 + Anti-miR-NC group,the si-HOXA-AS3 + Anti-miR-29 b colon cancer cell LOVO proliferation activity increased,the apoptosis rate decreased,the Bax protein expression in th
作者
游进
张佳
徐飞
王志伟
YOU Jin;ZHANG Jia;XU Fei;WANG Zhiwei(Department of Colorectal and Anal Surgery,Qingdao Central Hospital in Shandong Province,Qingdao,Shandong,266042)
出处
《实用临床医药杂志》
CAS
2022年第2期69-75,共7页
Journal of Clinical Medicine in Practice
基金
山东省医药卫生科技发展项目(2018WS360)。
