Impact of piR-35296 expression on proliferation in clear cell renal cell carcinoma
Objective To investigate the expression level of piR-35296 in clear cell renal cell carcinoma(ccRCC)and its pathological significance,as well as its effect on ccRCC cell prolifera-tion.Methods Surgical resection specimens and clinicopathological data from 67 ccRCC patients were collected.Differential expression profiles of piRNAs in ccRCC were detected using piRNA mi-croarray analysis,and validated by real-time quantitative reverse transcription-polymerase chain reac-tion(qRT-PCR).The effect of piR-35296 on the proliferative activity of ccRCC cells was examined by CCK-8 assay.Bioinformatic analysis predicted TAB2 as a potential downstream target gene of piR-35296;qRT-PCR was performed to detect theinfluence of piR-35296 overexpression or knockdown on TAB2 expression levels.Rescue experiments were conducted to verify the regulation of TAB2 by piR-35296 and its impact on ccRCC cell proliferation.The effect of piR-35296 knockdown on the growth of subcutaneous tumors in experimental nude mice was analyzed,and Ki-67 immunohistochemical staining was used to validate the influence of piR-35296 on ccRCC cell proliferation.Results Both piRNA expression profiling and qRT-PCR revealed an elevated expression level of piR-35296 in ccRCC(P＜0.05).The expression of piR-35296 was closely associated with tumor size and differentiation degree in ccRCC patients(x2=3.984,3.974,P＜0.05).CCK-8 proliferation assays demonstrated that piR-35296 overexpression enhanced the proliferative activity of ccRCC cells,whereas its down-regulation inhibited cell proliferation(P＜0.05).qRT-PCR experiments showed that TAB2 expres-sion was upregulated after piR-35296 overexpression and downregulated upon piR-35296 knockdown(P＜0.05).Rescue experiments indicated that piR-35296 promoted TAB2 mRNA and its protein expression in ccRCC cells,thereby stimulating cell proliferation;TAB2 downregulation inhibited ccRCC cell proliferation.In experimental nude mice,piR-35296 knockdown reduced the mass and-volume of subcutaneous tumors(P＜0.05).Immunohistochemical staining for Ki-67 revealed a de-creased percentage of Ki-67-positive cells after piR-35296 downregulation,suggesting reduced cell proliferation capacity(P＜0.05).Conclusion The piR-35296 serves as a novel oncogenic growth promoter in ccRCC.By elevating TAB2 expression,piR-35296 promotes the proliferative growth of ccRCC cells.
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