lncRNA LUCAT1通过调控miR-141-3p/SOX11轴促进肾透明细胞癌细胞的生长和转移的机制研究

Study on the mechanism of lncRNA LUCAT1 promoting the growth and metastasis of renal clear cell carcinoma by regulating the miR-141-3p/SOX11 axis

目的:探讨lncRNA LUCAT1通过调控miR-141-3p/SOX11轴对肾透明细胞癌细胞(kidney renal clear cell carcinoma,KIRC)的生长和转移的影响。方法:利用生信分析LUCAT1在KIRC中的表达,并预测下游的miRNA/mRNA调控轴;qRT-PCR用于检测LUCAT1、miR-141-3p和SOX11的表达,Western blot用于检测SOX11的表达;细胞生物学功能实验观察KIRC细胞的增殖、迁移和侵袭状况;双荧光素酶实验验证LUCAT1和miR-141-3p以及miR-141-3p和SOX11靶向结合关系;RIP实验验证LUCAT1和miR-141-3p的结合关系;裸鼠实验观察LUCAT1对肿瘤体内生长的影响。结果:通过生信分析和细胞实验发现LUCAT1和SOX11在KIRC中高表达,miR-141-3p在KIRC中低表达;MTT、伤口愈合和细胞侵袭实验结果显示LUCAT1促进KIRC细胞的增殖、迁移和侵袭。随后在双荧光素酶实验中显示LUCAT1可以作为miR-141-3p的分子海绵,同时我们还证实miR-141-3p能回复LUCAT1对KIRC细胞增殖、迁移和侵袭的促进作用。进一步研究发现SOX11是miR-141-3p的直接靶标,并且SOX11能回复miR-141-3p对KIRC细胞增殖、迁移和侵袭的抑制作用。最后我们还通过体内实验确定了LUCAT1能促进KIRC肿瘤的体内生长。结论:LUCAT1在KIRC中上调,并可以通过吸附miR-141-3p调控SOX11促进KIRC的生长和转移。这表明LUCAT1有望成为KIRC的生物标志物和潜在分子治疗靶点。

Objective:To investigate the effects of lncRNA LUCAT1 on the growth and metastasis of kidney renal clear cell carcinoma(KIRC) by regulating miR-141-3p/SOX11 axis.Methods:LUCAT1 expression in KIRC was analyzed by bioinformatics,and the downstream miRNA/mRNA regulation axis was predicted.qRT-PCR was used to detect the expression of LUCAT1,miR-141-3p and SOX11,and Western blot was used to detect the expression of SOX11.The proliferation,migration and invasion of KIRC cells were observed by cell biological function experiment.Dual luciferase assay verified the targeting binding relationship between LUCAT1,between miR-141-3p and miR-141-3p and SOX11.The binding relationship between LUCAT1 and miR-141-3p was verified by RIP experiments.The effect of LUCAT1 on tumor growth was observed in nude mice.Results:LUCAT1 and SOX11 were highly expressed in KIRC,while miR-141-3p was low expressed in KIRC by bioinformatics and cell expressions.Results of MTT,wound healing and cell invasion experiments showed that LUCAT1 promoted the proliferation,migration and invasion of KIRC cells.Subsequently,double luciferase experiments showed that LUCAT1 could act as a molecular sponge of miR-141-3p,and we also confirmed that miR-141-3p could restore the promoting effect of LUCAT1 on the proliferation,migration and invasion of KIRC cells.Further studies showed that SOX11 was a direct target of miR-141-3p,and SOX11 could restore the inhibitory effect of miR-141-3p on the proliferation,migration and invasion of KIRC cells.Finally,we also confirmed that LUCAT1 can promote the growth of KIRC tumors in vivo through in vivo experiments.Conclusion:LUCAT1 is upregulated in KIRC,and can promote the growth and metastasis of KIRC by regulating SOX11 through adsorption of miR-141-3p.This suggests that LUCAT1 is promising as a biomarker and potential molecular therapeutic target for KIRC.