SLC9A3-AS1 Regulates miR-148a-3p/ROCK1 Axis to Affect Biological Function of Renal Carcinoma Cells
Objective To examine lncRNA SLC9A3-AS1 expression levels within clear cell renal cell carcinoma(ccRCC)tis-sues and cells,aiming to elucidate its role in advancing the malignant progression of renal cancer cells.Methods GEPIA2 online software(http://gepia2.cancer-pku.cn/)was used to analyze the expression level of SLC9A3-AS1 in ccRCC tissues from the TCGA database.Else,real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was utilized to measure the ex-pression levels of SLC9A3-AS1 in various renal cancer cell lines as well as ccRCC and adjacent normal kidney tissues from 24 cases.To evaluate the impact of SLC9A3-AS1 on renal cancer cell proliferation and migration,CCK-8 cell proliferation assay and Trans well chamber migration assay were conducted.Western blot was employed to assess the expression levels of proteins relat-ed to proliferation and migration signaling pathways.Additionally,dual-luciferase reporter gene assay was conducted to validate the targeted regulatory relationship between SLC9A3-AS1 and the miR-148a-3p/ROCK1 axis.Results The GEPIA2 software analysis revealed a significant upregulation of SLC9A3-AS1 expression in ccRCC tissues compared to normal kidney tissues(P＜0.01).The qRT-PCR analysis showed that the expression of SLC9A3-AS1 was significantly upregulated in ccRCC tissues com-pared to adjacent normal kidney tissues in the 24 cases(P＜0.01).When compared to immortalized renal tubular epithelial cells,higher expressions of SLC9A3-AS1 were detected in four different types of renal cancer cell lines,with the most pronounced in-crease observed in 786-O cells(P＜0.01).Interfering with SLC9A3-AS1 expression had a notable inhibitory effect on the prolif-eration and migration of 786-O cells.This intervention led to an increase in E-cadherin protein expression and a decrease in N-cadherin and MMP2 protein expression levels(both P＜0.05).Likewise,the overexpression of miR-148a-3p exerted a similar in-hibitory effect on the proliferation and migration of 786-O cells(P＜0.01).The results of dual-luciferase reporter gene test sug-gested that SLC9A3-AS1 could specifically bind to miR-148a-3p,which further targeted the 3'UTR region of ROCK1 mR-NA.Overexpression of miR-148a-3p significantly downregulated the expression level of ROCK1 mRNA,while knockdown of miR-148a-3p had the opposite effect.The silence of SLC9A3-AS1 significantly suppressed the expression level of ROCK1 mR-NA and protein in 786-O cells(P＜0.01).Moreover,the data revealed that downregulation of miR-148a-3p could partially coun-teract the inhibitory effects of SLC9A3-AS1 silencing on the proliferation and migration of 786-O cells(P＜0.05).Conclusion SLC9A3-AS1 has a promoting effect on progression of ccRCC by modulating the miR-148a-3p/ROCK1 axis.It has the potential to serve as a novel molecular marker for ccRCC.
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