背景:透明细胞肾细胞癌(ccRCC)被确定为泌尿道中的恶性肿瘤。本研究试图探讨lncRNALINC00667在ccRCC发育中的生物学功能和分子机制。
方法:qRT-PCR监测LINC00667,miR-143-3p,和ZEB1水平。LINC00667,miR-143-3p,在ccRCC细胞中构建ZEB1过表达或敲低。细胞增殖,凋亡,迁移,并检测到细胞的侵袭。凋亡相关蛋白和上皮间质转化(EMT)相关蛋白的水平,和ZEB1通过WB检测。双荧光素酶报告基因测定和RNA下拉测定确定了LINC00667和miR-143-3p之间的结合关联,miR-143-3p和ZEB1。此外,裸鼠中的异种移植肿瘤模型用于评估体内肿瘤生长。
结果:LINC00667和ZEB1在ccRCC组织和细胞中显示高表达。miR-143-3p在ccRCC组织和细胞中低表达。LINC00667靶向和抑制miR-143-3p,以靶向方式抑制ZEB1表达。过表达LINC00667促进ccRCC细胞增殖,迁移,侵袭和EMT和延迟凋亡,而LINC00667敲低或miR-143-3p过表达发挥了相反的作用。拯救实验表明,过表达miR-143-3p抑制了LINC00667介导的致癌作用。过表达ZEB1减少miR-143-3p介导的肿瘤抑制作用。体内实验表明,LINC00667的过表达有助于ccRCC细胞的肿瘤生长,与miR-143-3p过表达相反,抑制了肿瘤的生长。
结论:LINC00667在ccRCC中上调,并通过靶向miR-143-3p增强ZEB1表达,进而加速ccRCC进展并诱导化学抗性。
Clear cell renal cell carcinoma (ccRCC) is identified as a malignant tumor in the urinary tract. The research was an attempt to probe the biological function and molecular mechanism of lncRNA LINC00667 in ccRCC development.
qRT-PCR monitored LINC00667, miR-143-3p, and ZEB1 levels. The models of LINC00667, miR-143-3p, and ZEB1 overexpression or knockdown were constructed in ccRCC cells. Cell proliferation, apoptosis, migration, and invasion of the cells were detected. The levels of apoptosis-associated proteins and epithelial-mesenchymal transition (EMT)-related proteins, and ZEB1 were detected by WB. Dual-luciferase reporter assay and RNA pull-down assay identified the binding association between LINC00667 and miR-143-3p, miR-143-3p and ZEB1. Moreover, a xenograft tumor model in nude mice was used for evaluating tumor growth in vivo.
LINC00667 and ZEB1 displayed high expression in ccRCC tissues and cells. miR-143-3p was lowly expressed in ccRCC tissues and cells. LINC00667 targeted and repressed miR-143-3p, which inhibited ZEB1 expression in a targeted manner. Overexpression of LINC00667 facilitated ccRCC cell proliferation, migration, invasion and EMT and retarded apoptosis, whereas LINC00667 knockdown or miR-143-3p overexpression exerted reverse effects. The rescue experiments indicated that overexpressing miR-143-3p dampened LINC00667-mediated oncogenic effects. Overexpressing ZEB1 diminished miR-143-3p-mediated tumor-suppressive effects. In-vivo experiments displayed that overexpression of LINC00667 contributed to the tumor growth of ccRCC cells, in contrast to miR-143-3p overexpression, which restrained the tumor growth.
LINC00667 is up-regulated in ccRCC and enhances the ZEB1 expression by targeting miR-143-3p, which in turn accelerates ccRCC progression and induces chemoresistance.