Department of Hepatobiliary and Pancreas Surgery, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
Teacher's Office of Clinical & Medical Nursing, Changchun Medical College, Changchun, Jilin, China.
J Cell Mol Med. 2018 Jul;22(7):3434-3441. doi: 10.1111/jcmm.13620. Epub 2018 Apr 24.
In this study, we investigated how miR-10b-3p regulated the proliferation, migration, invasion in hepatocellular carcinoma (HCC) at both in vitro and in vivo levels. CMTM5 was among the differentially expressed genes (data from TCGA). The expression of miR-10b-3p and CMTM5 was detected by qRT-PCR and Western blot (WB). TargetScan was used to acquire the binding sites. Dual-luciferase reporter gene assay was used to verify the direct target relationship between miR-10b-3p and CMTM5. WB analysis proved that miR-10b-3p suppressed CMTM5 expression. Furthermore, proliferation, invasion and migration of HCC cells were measured by MTT assay, colony formation assay, transwell assay and wound-healing assay, respectively. Kaplan-Meier plotter valued the overall survival of CMTM5. Finally, xenograft assay was also conducted to verify the effects of miR-10b-3p/CMTM5 axis in vivo. Up-regulation of miR-10b-3p and down-regulation of CMTM5 were detected in HCC tissues and cell lines. CMTM5 was verified as a target gene of miR-10b-3p. The overexpression of CMTM5 contributed to the suppression of the proliferative, migratory and invasive abilities of HCC cells. Moreover, the up-regulation of miR-10b-3p and down-regulation of CMTM5 were observed to be associated with worse overall survival. Lastly, we have confirmed the carcinogenesis-related roles of miR-10b-3p and CMTM5 in vivo. We concluded that the up-regulation of miR-10b-3p promoted the progression of HCC cells via targeting CMTM5.
在这项研究中,我们研究了 miR-10b-3p 如何在体内和体外水平调节肝癌(HCC)的增殖、迁移和侵袭。CMTM5 是差异表达基因之一(来自 TCGA 的数据)。通过 qRT-PCR 和 Western blot(WB)检测 miR-10b-3p 和 CMTM5 的表达。使用 TargetScan 获取结合位点。双荧光素酶报告基因检测用于验证 miR-10b-3p 和 CMTM5 之间的直接靶标关系。WB 分析证实 miR-10b-3p 抑制 CMTM5 表达。此外,通过 MTT 测定、集落形成测定、Transwell 测定和划痕愈合测定分别测量 HCC 细胞的增殖、侵袭和迁移。Kaplan-Meier plotter 评估了 CMTM5 的总生存率。最后,还进行了异种移植实验以验证 miR-10b-3p/CMTM5 轴在体内的作用。在 HCC 组织和细胞系中检测到 miR-10b-3p 的上调和 CMTM5 的下调。CMTM5 被验证为 miR-10b-3p 的靶基因。CMTM5 的过表达抑制 HCC 细胞的增殖、迁移和侵袭能力。此外,miR-10b-3p 的上调和 CMTM5 的下调与较差的总生存率相关。最后,我们在体内证实了 miR-10b-3p 和 CMTM5 的致癌作用。我们得出结论,miR-10b-3p 的上调通过靶向 CMTM5 促进 HCC 细胞的进展。