Poor expression of microRNA-135b results in the inhibition of cisplatin resistance and proliferation and induces the apoptosis of gastric cancer cells through MST1-mediated MAPK signaling pathway
Poor expression of microRNA-135b results in the inhibition of cisplatin resistance and proliferation and induces the apoptosis of gastric cancer cells through MST1-mediated MAPK signaling pathway
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microRNA-135b低表达通过MST1介导的MAPK信号通路抑制顺铂耐药和增殖并诱导胃癌细胞凋亡
DOI：
10.1096/fj.201800618rrr
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发表时间：
2019-03-01
期刊：
影响因子：
4.8
通讯作者：
Chen，Qing
中科院分区：
文献类型：
--
作者：
Zhou，Jie;Chen，Qing
胃癌(GC)已被列为全球癌症相关死亡的第四大原因。由于能够抑制靶基因的表达，microRNAs(MiRNAs)被认为是参与肿瘤形成和发展的关键因素之一。因此，本研究通过MAPK信号通路，靶向哺乳动物Ste20-like kinase1(MST1)，研究microRNA-135B(miR-135B)对胃癌细胞顺铂[顺铂]耐药性的影响。采用基因表达芯片技术筛选与胃癌相关的差异表达基因。3-[4，5-二甲基噻唑-2-基]-2，5-二苯基四氮唑溴化法测定GC细胞对顺铂的敏感性。利用生物信息学数据库和双荧光素酶报告基因分析验证MST1是否是miR-135B的直接靶基因。制备对顺铂耐药的胃癌细胞系，分别对其进行培养和转染，然后将其接种到裸鼠体内，再用顺铂处理，以探讨miR-135B与MST1在胃癌发生发展中的作用机制。这些结果高度表明MST1在GC的发生和GC对CDDP的敏感性中所起的关键作用。MIR-135B被发现调节MST1，MST1反过来对GC的发展产生影响。MKN28对CDDP最敏感，而MKN45对CDDP最不敏感。此外，miR-135B表达下调导致MAPK信号通路失活，MST1和Bax表达增加，p-p38MAPK、p-ERK1/2、P-糖蛋白、p38MAPK、ERK1/2、多药耐药蛋白1、多药耐药相关蛋白1、肺耐药相关蛋白和Bcl-2表达降低，从而抑制了GC细胞对顺铂的耐药性。MiR-135B的下调也抑制了细胞的增殖，诱导了GC细胞的凋亡率。综上所述，本研究结果表明miR-135B的下调可诱导胃癌细胞的凋亡，并通过失活MAPK信号通路和上调MST1的表达来抑制胃癌细胞的增殖和顺铂耐药。
Gastric cancer (GC) has been classified as the fourth leading cause of cancer-related deaths worldwide. Due to their ability to suppress the expression of target genes, microRNAs (miRNAs) are listed as one of the key elements involved in the formation and development of tumors. This study was therefore conducted to investigate the effects of microRNA-135b (miR-135b) on cisplatin [cis-diamminedichloroplatinum (CDDP)] resistance of GC cells through the MAPK signaling pathway by targeting mammalian ste20-like kinase 1 (MST1). A microarray-based gene expression analysis was performed to screen the GC-related differentially expressed genes. The 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay was performed to determine the sensitivity of GC cells to CDDP. The bioinformatics database and dual luciferase reporter gene assay were used to check whether MST1 was a direct target gene of miR-135b. GC cell lines were prepared with high CDDP resistance, after which they were cultured and transfected respectively, followed by the administration of transfected cells into nude mice and subsequent treatment with CDDP in an attempt to identify the underlying mechanisms and functions of miR-135b in relation to MST1 in GC progression. The results were highly indicative of the crucial role played by MST1 in the development of GC and the sensitivity of GC to CDDP. miR-135b was found to regulate MST1, which in turn had an impact on the development of GC. MKN28 was observed to be most sensitive to CDDP, whereas MKN45 presented with the poorest sensitivity to CDDP. Furthermore, the down-regulation of miR-135b resulted in inactivation of the MAPK signaling pathway; increased the expression of MST1 and Bax; and decreased expression of p-p38MAPK, p-ERK1/2, P-glycoprotein, p38MAPK, ERK1/2, multidrug resistance protein 1, multidrug resistance–associated protein 1, lung resistance–related protein, and Bcl-2, thus inhibiting CDDP resistance of GC cells. The down-regulation of miR-135b also restrained cell proliferation and induced the apoptosis rate of GC cells. In summary, the results of this study showed that the down-regulation of miR-135b induced apoptosis, and it inhibited proliferation and CDDP resistance of GC cells by inactivating the MAPK signaling pathway and increasing the expression of MST1.—Zhou, J., Chen, Q. Poor expression of microRNA-135b results in the inhibition of cisplatin resistance and proliferation and induces the apoptosis of gastric cancer cells through MST1-mediated MAPK signaling pathway.