Yang Fan, Wu Qingjian, Zhang Le, Xie Wei, Sun Xiaoli, Zhang Yan, Wang Lei, Dai Qian, Yu Hua, Chen Qian, Sheng Halei, Qiu Jing, He Xiaomei, Miao Hongming, He Fengtian, Zhang Kebin
Central Laboratory, Xinqiao Hospital, Army Medical University (Third Military Medical University), Chongqing 400037, China.
Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Army Medical University (Third Military Medical University), Chongqing 400038, China.
J Cancer. 2019 Aug 19;10(19):4662-4670. doi: 10.7150/jca.29280. eCollection 2019.
: Long noncoding RNAs (lncRNAs) have been demonstrated to play essential roles in renal cell carcinoma (RCC). However, the role of lncRNA KCNQ1DN in RCC remains unclear. : The expression of KCNQ1DN in RCC and the corresponding adjacent tissues was measured by qPCR. RNA fluorescence in situ hybridization (FISH) assay, methylation analysis, reporter gene assays and functional tests were performed to reveal the effects of KCNQ1DN on RCC. : In the present study, we found that lncRNA KCNQ1DN was notably decreased in RCC tissues and cell lines. RNA FISH assay showed that KCNQ1DN mainly localized to the cytoplasm. Methylation analysis revealed that the proximal region of KCNQ1DN promoter was hypermethylated in RCC tissues relative to the adjacent normal ones. Functional studies clarified that KCNQ1DN repressed the RCC cell growth and cell cycle progression. Mechanistically, KCNQ1DN inhibited the expression of c-Myc, which might further upregulate cyclin D1 and suppress p27 at mRNA and protein levels in RCC cells. Reporter gene assays revealed that the transcriptional activity of promoter was inhibited by KCNQ1DN. The experiments in nude mice showed that KCNQ1DN overexpression dramatically repressed the growth of xenograft tumors and the expression of corresponding c-Myc. : These results indicated that KCNQ1DN inhibit the growth of RCC cells and through repressing the oncogene suggesting that KCNQ1DN may serve as a novel target for the treatment of RCC.
长链非编码RNA(lncRNAs)已被证明在肾细胞癌(RCC)中发挥重要作用。然而,lncRNA KCNQ1DN在RCC中的作用仍不清楚。
通过qPCR检测KCNQ1DN在RCC及相应癌旁组织中的表达。进行RNA荧光原位杂交(FISH)分析、甲基化分析、报告基因检测和功能测试,以揭示KCNQ1DN对RCC的影响。
在本研究中,我们发现lncRNA KCNQ1DN在RCC组织和细胞系中显著降低。RNA FISH分析表明KCNQ1DN主要定位于细胞质。甲基化分析显示,相对于相邻正常组织,RCC组织中KCNQ1DN启动子的近端区域发生高甲基化。功能研究表明,KCNQ1DN抑制RCC细胞生长和细胞周期进程。机制上,KCNQ1DN抑制c-Myc的表达,这可能进一步上调细胞周期蛋白D1并在mRNA和蛋白质水平上抑制RCC细胞中的p27。报告基因检测显示,KCNQ1DN抑制启动子的转录活性。裸鼠实验表明,KCNQ1DN过表达显著抑制异种移植肿瘤的生长及相应c-Myc的表达。
这些结果表明,KCNQ1DN通过抑制癌基因c-Myc来抑制RCC细胞生长,提示KCNQ1DN可能成为RCC治疗的新靶点。