论文部分内容阅读
目的探讨miR-24在结肠癌中发挥的作用并研究其是否和多柔比星的体外治疗有关。方法用RT-q PCR法检测miR-24在结肠癌患者血浆中及结肠癌细胞系中的表达水平。MTT法检测miR-24反义核酸对多柔比星杀伤结肠癌细胞能力的影响。利用生物信息学、定量PCR及Western blot法验证miR-24是否调节结肠癌细胞BIM的表达。运用JC-1染色、Annexin V染色及Western blot法研究miR-24反义核酸影响多柔比星疗效的信号通路。结果结肠癌患者血浆及结肠癌细胞系中miR-24表达水平显著升高。miR-24反义核酸可显著增强多柔比星对SW480细胞的杀伤活性。定量PCR及Western blot实验表明miR-24的靶基因可能为BIM。miR-24反义核酸联合多柔比星可引起SW480细胞线粒体膜电位的丧失并诱导线粒体内Smac/DIABLO的释放,进而引起caspase-3的活化和凋亡的发生。转染BIM si RNA后miR-24反义核酸联合多柔比星对SW480细胞的凋亡诱导效应显著降低。结论 Mi R-24反义核酸通过BIM-Smac/Diablo途径促进多柔比星对结肠癌细胞的凋亡诱导效应。
Objective To investigate the role of miR-24 in colon cancer and investigate whether it is related to the in vitro treatment of doxorubicin. Methods RT-qPCR method was used to detect the expression of miR-24 in the plasma of colon cancer patients and colon cancer cell lines. Effect of miR-24 antisense oligonucleotide on the ability of doxorubicin to kill colon cancer cells by MTT assay. Bioinformatics, quantitative PCR and Western blot were used to verify whether miR-24 regulates BIM expression in colon cancer cells. JC-1 staining, Annexin V staining and Western blot assay miR-24 antisense nucleic acid signaling pathway affecting the efficacy of doxorubicin. Results The expression of miR-24 in plasma and colon cancer cell lines was significantly increased in colon cancer patients. miR-24 antisense nucleic acid can significantly enhance the doxorubicin on SW480 cell killing activity. Quantitative PCR and Western blot showed that miR-24 target gene may be BIM. The combination of miR-24 antisense and doxorubicin can induce the loss of mitochondrial membrane potential and induce the release of Smac / DIABLO in SW480 cells, leading to the activation and apoptosis of caspase-3. The apoptosis-inducing effect of miR-24 antisense nucleic acid combined with doxorubicin on SW480 cells was significantly reduced after BIM si RNA transfection. Conclusion Mi R-24 antisense oligonucleotide promotes the apoptosis-inducing effect of doxorubicine on colon cancer cells through the BIM-Smac / Diablo pathway.