下调长链非编码RNA <i>HOXA-AS3</i>靶向促进微小RNA-29b对结肠癌细胞LOVO凋亡和奥沙利铂敏感性的影响

游进; 张佳; 徐飞; 王志伟

doi:10.7619/jcmp.20213051

下调长链非编码RNA HOXA-AS3靶向促进微小RNA-29b对结肠癌细胞LOVO凋亡和奥沙利铂敏感性的影响

Effect of down-regulating lncRNA HOXA-AS3 targeted promotion of miR-29b on the apoptosis of colon cancer cells LOVO and the sensitivity of oxaliplatin

摘要

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凋亡并提高奥沙利铂敏感性。

Abstract:
Objective To investigate effect of down-regulating lncRNA HOXA-AS3 targeted promotion of miR-29b 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-29b group, L-OHP+si-HOXA-AS3 +Anti-miR-NC group and L-OHP+si-HOXA-AS3 +Anti-miR-29b 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-29b.
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-29b colon cancer cell LOVO proliferation activity increased, the apoptosis rate decreased, the Bax protein expression in the cells decreased, and Bcl-2 protein expression increased (P < 0.05). Compared with the L-OHP+si-HOXA-AS3 +Anti-miR-NC group, the colon cancer cell LOVO proliferation activity in the L-OHP+si-HOXA-AS3 +Anti-miR-29b group increased, and the apoptosis rate decreased, the expression of Bax protein in cells decreased, and the expression of Bcl-2 protein increased (P < 0.05). Down-regulation of HOXA-AS3 targeted to promote miR-29b expression.
Conclusion Down-regulation of HOXA-AS3 targeting promotes miR-29b to induce the apoptosis of colon cancer cells LOVO and increase the sensitivity of oxaliplatin.

HTML全文

参考文献(21)

施引文献

资源附件(0)