School of Basic Medicine, Youjiang Medical University for Nationalities, Baise, Guangxi Province, China.
Eur Rev Med Pharmacol Sci. 2020 Jul;24(14):7645-7654. doi: 10.26355/eurrev_202007_22265.
To investigate the expression of Long non-coding RNA ADIPOQ and its facilitating effects on proliferation and invasion of colorectal cancer by modulating the expression of TP53 via sponging with miR-219c-3p.
qRT-PCR was performed to detect the expressions of ADIPOQ and TP53 in human colorectal cancer tissues and cells. CCK-8 assay was performed to evaluate the Caco-2 cells proliferation and transwell assay was performed to evaluate the Caco-2 cells migration. The relationship between ADIPOQ and miR-219c-3p was detected by statistical analysis. Target prediction and Luciferase activity assay were conducted to investigate the binding site and interaction between ADIPOQ and miR-219c-3p. Further, we cloned the mice TP53 3'-UTR into the Luciferase reporter vector and constructed miR-219c-3p binding mutants to verify the inhibited regulation of miR-219c-3p to the TP53 expression.
The results suggested that the expression of ADIPOQ and TP53 was downregulated in human colorectal cancer tissues and Caco-2 cells. qRT-PCR and CCK-8 assay showed that ADIPOQ expression is correlated with the proliferation of colorectal cancer cells. Transwell assay showed that ADIPOQ regulated the migration ability of colorectal cancer cells. The bioinformatics prediction and Luciferase assay demonstrated that ADIPOQ serves as ceRNA for miR-219c-3p to further regulate the expression of TP53.
For the first time, we found that lncRNA-ADIPOQ was downregulated in human colorectal cancer cells, which could facilitate tumor proliferation, migration and invasion as a ceRNA by sponging with miR-219c-3p.
通过与 miR-219c-3p 海绵吸附作用调节 TP53 的表达,研究长链非编码 RNA ADIPOQ 的表达及其促进结直肠癌细胞增殖和侵袭的作用。
qRT-PCR 检测人结直肠癌细胞和组织中 ADIPOQ 和 TP53 的表达。CCK-8 检测 Caco-2 细胞增殖,Transwell 检测 Caco-2 细胞迁移。统计分析检测 ADIPOQ 与 miR-219c-3p 的关系。通过靶基因预测和荧光素酶活性测定分析 ADIPOQ 与 miR-219c-3p 的结合位点和相互作用。进一步将小鼠 TP53 3'-UTR 克隆到荧光素酶报告载体中,并构建 miR-219c-3p 结合突变体,验证 miR-219c-3p 对 TP53 表达的抑制调节作用。
结果表明,ADIPOQ 和 TP53 的表达在人结直肠癌细胞和 Caco-2 细胞中下调。qRT-PCR 和 CCK-8 检测表明 ADIPOQ 表达与结直肠癌细胞增殖相关。Transwell 检测表明 ADIPOQ 调节结直肠癌细胞的迁移能力。生物信息学预测和荧光素酶测定表明,ADIPOQ 作为 miR-219c-3p 的 ceRNA,进一步调节 TP53 的表达。
我们首次发现长链非编码 RNA ADIPOQ 在人结直肠癌细胞中下调,作为 ceRNA 通过与 miR-219c-3p 海绵吸附作用促进肿瘤增殖、迁移和侵袭。