内脂素通过下调SDF-1/CXCR4介导的miR-140-3p表达促进结肠癌细胞增殖和侵袭的作用。

Role of visfatin in promoting proliferation and invasion of colorectal cancer cells by downregulating SDF-1/CXCR4-mediated miR-140-3p expression.

作者信息

Zhao Q, Li J-Y, Zhang J, Long Y-X, Li Y-J, Guo X-D, Wei M-N, Liu W-J

机构信息

Department of General Surgery Ⅰ, the First People's Hospital of Yunnan Province, the Affiliated Hospital of Kunming University of Science and Technology, Kunming, Yunnan Province, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 May;24(10):5367-5377. doi: 10.26355/eurrev_202005_21320.

DOI:10.26355/eurrev_202005_21320

PMID:32495871

Abstract

OBJECTIVE

Visfatin is significantly upregulated in colorectal cancer (CRC). However, its exact role in CRC progression and the regulatory mechanism involved in this process have not been fully illuminated. The aim of this study was to investigate the roles of visfatin in CRC progression and the potential molecular mechanism.

MATERIALS AND METHODS

In vitro, two CRC cell lines (DLD-1 and SW480) were transfected with visfatin, si-visfatin, and their control vectors. Some cells were transfected with miR-140-3p mimics or miRNA negative control. Cell Counting Kit-8 and transwell invasive assays were used to detect cell proliferation and invasion ability. Luciferase reporter assays were performed to confirm whether CXC motif chemokine receptor 4 (CXCR4) directly targets miR-140-3p. Western blotting and qRT-PCR analyses were respectively conducted to evaluate the protein and mRNA levels of stromal cell-derived factor-1 (SDF-1) and CXCR4. In vivo, DLD-1 cells transfected with visfatin construct or vector control were inoculated into nude mice. After 5 weeks, the mice were sacrificed, and the tumor nodules were weighed. The expression of visfatin, SDF-1, and CXCR4 in tumor tissues was detected via immunohistochemistry analysis.

RESULTS

In vitro, the transfection of visfatin promoted the proliferation and invasion of CRC cells, as well as upregulated the expression of SDF-1/CXCR4. MiR-140-3p directly targets the 3'untranslated region of CXCR4. MiR-140-3p expression was downregulated by treatment with visfatin, and miR-140-3p exerted similar effects to those of visfatin knockdown on the proliferation and invasion of CRC cells. In vivo, visfatin stimulated CRC tumor growth and downregulated miR-140-3p expression, whereas it upregulated SDF-1/CXCR4 expression.

CONCLUSIONS

Visfatin promotes CRC progression by downregulating the SDF-1/CXCR4-mediated expression of miR-140-3p both in vitro and in vivo.

摘要

目的

内脂素在结直肠癌（CRC）中显著上调。然而，其在CRC进展中的具体作用以及该过程中涉及的调控机制尚未完全阐明。本研究旨在探讨内脂素在CRC进展中的作用及潜在分子机制。

材料与方法

在体外，用内脂素、内脂素小干扰RNA（si-visfatin）及其对照载体转染两种CRC细胞系（DLD-1和SW480）。部分细胞用miR-140-3p模拟物或微小RNA阴性对照转染。采用细胞计数试剂盒-8和Transwell侵袭实验检测细胞增殖和侵袭能力。进行荧光素酶报告基因实验以确认CXC基序趋化因子受体4（CXCR4）是否直接靶向miR-140-3p。分别进行蛋白质免疫印迹法和定量逆转录聚合酶链反应分析以评估基质细胞衍生因子-1（SDF-1）和CXCR4的蛋白质和信使核糖核酸水平。在体内，将用内脂素构建体或载体对照转染的DLD-1细胞接种到裸鼠体内。5周后，处死小鼠，称取肿瘤结节重量。通过免疫组织化学分析检测肿瘤组织中内脂素、SDF-1和CXCR4的表达。

结果

在体外，内脂素转染促进了CRC细胞的增殖和侵袭，并上调了SDF-1/CXCR4的表达。MiR-140-3p直接靶向CXCR4的3'非翻译区。内脂素处理下调了miR-140-3p的表达，且miR-140-3p对CRC细胞增殖和侵袭的影响与内脂素敲低相似。在体内，内脂素刺激CRC肿瘤生长并下调miR-140-3p表达，而上调SDF-1/CXCR4表达。