Department of Biochemistry and Molecular Biology, School of Basic Medicine, and Huazhong University of Science and Technology, Wuhan, China.
Department of Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
FASEB J. 2019 Mar;33(3):3420-3436. doi: 10.1096/fj.201800618RRR. Epub 2018 Dec 21.
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.
胃癌 (GC) 已被列为全球癌症相关死亡的第四大主要原因。由于它们能够抑制靶基因的表达,微小 RNA(miRNA)被列为参与肿瘤形成和发展的关键因素之一。因此,本研究通过靶向哺乳动物 Ste20 样激酶 1(MST1),通过 MAPK 信号通路研究了 microRNA-135b(miR-135b)对 GC 细胞顺铂 [顺式二氨二氯铂 (CDDP)] 耐药性的影响。进行了基于微阵列的基因表达分析,以筛选与 GC 相关的差异表达基因。使用 3-[4,5-二甲基噻唑-2-基]-2,5 二苯基四氮唑溴盐 (MTT) 测定法测定 GC 细胞对 CDDP 的敏感性。生物信息学数据库和双荧光素酶报告基因测定用于检查 MST1 是否是 miR-135b 的直接靶基因。制备具有高 CDDP 耐药性的 GC 细胞系,然后分别培养和转染,然后将转染细胞注入裸鼠,并随后用 CDDP 处理,试图确定 miR-135b 在 GC 进展中与 MST1 相关的潜在机制和功能。结果高度表明 MST1 在 GC 的发展和 GC 对 CDDP 的敏感性中起着至关重要的作用。发现 miR-135b 调节 MST1,进而影响 GC 的发展。观察到 MKN28 对 CDDP 最敏感,而 MKN45 对 CDDP 的敏感性最差。此外,下调 miR-135b 导致 MAPK 信号通路失活;增加 MST1 和 Bax 的表达;并降低 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 的表达,从而抑制 GC 细胞对 CDDP 的耐药性。miR-135b 的下调还抑制了 GC 细胞的增殖并诱导了其凋亡率。总之,本研究结果表明,下调 miR-135b 通过失活 MAPK 信号通路和增加 MST1 的表达来诱导 GC 细胞凋亡,并抑制 GC 细胞的增殖和 CDDP 耐药性。