miR-134-5p靶向MRPL58调控乳腺癌细胞MCF7的凋亡研究

目的:探讨miR-134-5p靶向MRPL58调控乳腺癌细胞MCF7凋亡的潜在分子机制。方法:通过高通量测序检测正常乳腺上皮细胞MCF10A与乳腺癌细胞MCF7中的差异miRNA。在乳腺癌细胞MCF7中过表达差异倍数大于7的miRNA,检测其凋亡水平。通过miRDB在线分析miRNA潜在的靶标基因。通过过表达或敲低miRNA和荧光素酶报告实验确定miRNA的靶标基因。结果:正常乳腺上皮细胞MCF10A与乳腺癌细胞MCF7的总miRNA高通量测序结果发现,MCF10A细胞中miRNA比乳腺癌细胞MCF7表达量大于7倍的有30种。过表达上述miRNA后,miR-134-5p增强了乳腺癌细胞MCF7的凋亡水平(P<0.05)。敲低miR-134-5p后,乳腺癌细胞MCF7的凋亡水平下降(P<0.05)。miR-134-5p在癌旁组织中的表达水平高于乳腺癌组织(P<0.05)。过表达miR-134-5p后,KIAA0040,ZMAT5,RAB27A,TESMIN,MRPL58,ZFPM2,NUP98的表达水平下降(P<0.05)。敲低上述基因后,敲低MRPL58时乳腺癌细胞MCF7的凋亡水平上升(P<0.05)。过表达MRPL58后,乳腺癌细胞MCF7的凋亡水平下降。敲低miR-134-5p后,MRPL58的表达水平上升。过表达miR-134-5p后,MRPL58的表达水平下降。同时,miR-134-5p靶向MRPL58的3端非编码区(P<0.05)。同时敲低miR-134-5p和MRPL58后,乳腺癌细胞MCF7的凋亡水平没有显著改变(P>0.05)。同时过表达miR-134-5p和MRPL58后,乳腺癌细胞MCF7的凋亡水平没有显著改变(P>0.05)。结论:miR-134-5p通过靶向MRPL58 mRNA的3端非编码区以降低MRPL58的翻译水平,最终促进了乳腺癌细胞MCF7的凋亡。

Study on miR-134-5p targeting MRPL58 to regulate the apoptosis of breast cancer cell MCF7

Objective:To investigate the potential molecular mechanism of miR-134-5p targeting MRPL58 to regulate breast cancer cell MCF7 apoptosis.Methods:High-throughput sequencing was used to detect the differential miRNAs in normal breast epithelial cells MCF10A and breast cancer cells MCF7.Overexpression of miRNAs with a differential multiple of greater than 7 in breast cancer cell MCF7,and detect their apoptosis levels.Online analysis of miRNA potential target genes through miRDB.Determine the target gene of miRNA by overexpression or knockdown of miRNA and luciferase reporter experiment.Results:High-throughput sequencing of total miRNA in normal breast epithelial cells MCF10A and breast cancer cells MCF7 showed that there were 30 miRNAs in MCF10A cells that expressed more than 7 times higher than breast cancer cells MCF7.After overexpressing the above miRNA,miR-134-5p enhanced the apoptosis level of breast cancer cell MCF7(P<0.05).After knocking down miR-134-5p,the apoptosis level of breast cancer cell MCF7 decreased(P<0.05).In addition,the expression level of miR-134-5p in adjacent tissues was higher than that in breast cancer tissues(P<0.05).After overexpression of miR-134-5p,the expression levels of KIAA0040,ZMAT5,RAB27A,TESMIN,MRPL58,ZFPM2,and NUP98 were found to decrease(P<0.05).After knocking down the above genes,the apoptosis level of breast cancer cell MCF7 increased when MRPL58 was knocked down(P<0.05).After overexpression of MRPL58,the apoptosis level of breast cancer cell MCF7 decreased.After knocking down miR-134-5p,the expression level of MRPL58 increased.After overexpression of miR-134-5p,the expression level of MRPL58 decreased.At the same time,miR-134-5p targeted the 3rd non-coding region of MRPL58(P<0.05).After knocking down miR-134-5p and MRPL58 at the same time,the apoptosis level of breast cancer cell MCF7 did not change significantly(P>0.05).After overexpression of miR-134-5p and MRPL58 at the same time,the apoptosis level of breast cancer cell MCF7 did not change significantly(P>0.05).Conclusion:miR-134-5p reduces the translation level of MRPL58 by targeting the 3 non-coding region of MRPL58 mRNA,and ultimately promotes the apoptosis of breast cancer cell MCF7.