透明细胞肾细胞癌中piR-35296表达对增殖的影响
Impact of piR-35296 expression on proliferation in clear cell renal cell carcinoma
胡道吉 1王喻烨 2王绪 2严雪冰 2王成海3
作者信息
- 1. 扬州大学医学院,江苏扬州,225009;江苏省沭阳县中医院肾内科,江苏沭阳,223600
- 2. 扬州大学医学院,江苏扬州,225009
- 3. 扬州大学医学院,江苏扬州,225009;江苏省中西医结合老年病防治重点实验室,江苏扬州,225009
- 折叠
摘要
目的 研究透明细胞肾细胞癌(ccRCC)中piR-35296的表达水平与病理意义以及对ccRCC细胞增殖的影响.方法 收集67例ccRCC患者手术切除标本及临床病理资料.通过piRNA基因芯片检测ccRCC中piRNA差异性表达谱,并用实时荧光定量聚合酶链反应(qRT-PCR)实验对差异性表达的piRNA进行验证.通过CCK-8实验检测piR-35296对ccRCC细胞增殖活性的影响.通过生物信息学预测TAB2是piR-35296的下游靶基因;通过qRT-PCR实验检测过表达piR-35296或敲低piR-35296表达对TAB2表达水平的影响.通过Rescue实验验证piR-35296对TAB2的调控以及对ccRCC细胞增殖的影响.分析实验性裸鼠实验中敲低piR-35296表达对皮下移植瘤生长的影响;通过增殖指标Ki-67免疫组化染色验证piR-35296对ccRCC细胞增殖的影响.结果 piRNA表达谱和qRT-PCR均检测出piR-352961在ccRCC中表达水平升高(P＜0.05).piR-35296的表达与ccRCC患者肿瘤大小和分化程度密切相关(x2=3.984、3.974,P＜0.05).CCK-8增殖实验结果表明,过表达piR-35296能提高ccRCC细胞的增殖活性,降低piR-35296表达可抑制ccRCC细胞的增殖活性(P＜0.05).qRT-PCR实验发现,过表达piR-35296后TAB2表达上调,降低piR-35296表达后TAB2表达下调(P＜0.05).Rescue实验提示,piR-35296能促进ccRCC细胞的TAB2 mRNA及其蛋白表达,并促进ccRCC细胞增殖;降低TAB2表达可抑制ccRCC细胞增殖.实验性裸鼠实验中,降低piR-35296表达后,移植瘤质量和体积减小(P＜0.05).免疫组化染色Ki-67结果显示,降低piR-35296表达后,Ki-67阳性细胞比率降低,提示细胞增殖能力下降(P＜0.05).结论 在ccRCC中,piR-35296是一个新的促癌基因.piR-35296可通过提高TAB2的表达水平,促进ccRCC细胞的增殖.
Abstract
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.
关键词
肾细胞癌/透明细胞肾细胞癌/piR-35296/TAB2表达水平/细胞增殖Key words
renal cell carcinoma/clear cell renal cell carcinoma/piR-35296/TAB2 expres-sion level/cell proliferation引用本文复制引用
基金项目
江苏省卫生健康委科研基金资助项目(M2020024)
出版年
2024
国家科技期刊平台
NSTL
万方数据