MP16-05 LOWERING MRNA EXPRESSION OF PLEKHO1 INHIBITS TUMOR GROWTH OF RCC CELLS IN VITRO AND IN VIVO, POTENTIALLY THROUGH MODULATING THE HIPPO AND MAPK/JNK SIGNALLING PATHWAYS

You have accessJournal of UrologyKidney Cancer: Basic Research & Pathophysiology I (MP16)1 Apr 2019MP16-05 LOWERING MRNA EXPRESSION OF PLEKHO1 INHIBITS TUMOR GROWTH OF RCC CELLS IN VITRO AND IN VIVO, POTENTIALLY THROUGH MODULATING THE HIPPO AND MAPK/JNK SIGNALLING PATHWAYS Yu Zeng*, Zi Yu, Shui Fu, Gejun Zhang, Chengcheng Lv, Qingzhuo Dong, Chuize Kong, and Cheng Fu Yu Zeng*Yu Zeng* More articles by this author , Zi YuZi Yu More articles by this author , Shui FuShui Fu More articles by this author , Gejun ZhangGejun Zhang More articles by this author , Chengcheng LvChengcheng Lv More articles by this author , Qingzhuo DongQingzhuo Dong More articles by this author , Chuize KongChuize Kong More articles by this author , and Cheng FuCheng Fu More articles by this author View All Author Informationhttps://doi.org/10.1097/01.JU.0000555341.83310.20AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVES: Renal cell carcinoma (RCC) is the most common type of kidney cancer word widely. Although target therapy and checkpoint immunotherapy improve the survival of metastatic RCC, a large number of patients are still incurable. METHODS: The raw RNA-sequencing data of RCC were download from the Cancer Genome Atlas (TCGA) database and exported by using R-project. A high-content cell viability assay was used to screen genes those are potentially regulating cell proliferation after downregulating the mRNA expressions of each gene by shRNA. RT-PCR was conducted to detect the mRNA expression of pleckstrin homology domain containing O1 (PLEKHO1) gene in 30 paired localized tumor and para-tumor kidney tissue samples. The effects of lowering the expression of PLEKHO1 were assessed by CCK-8 assay, Celigo assay, flow cytometry and transwell assay, separately. Additionally, xenograft tumor models were established in nude mice to investigate the function of PLEKHO1 in vivo. Gene expression microarray and co-expression analysis followed by Western blot were further conducted to explore the involvement of PLEKHO1 in signalling pathways. RESULTS: Sixteen genes were identified to be consistently highly expressed in ccRCC tissue compared with the matched para-tumor tissue in TCGA data set, while downregulation of only two of those significantly inhibited the viability of RCC cells. Among two genes, PLEKHO1 was unknown in its relevance to RCC and its expression level was shown to be associated with prognosis of RCC in the TCGA data set. Next, upregulation of PLEKHO1 mRNA was confirmed in RCC tissue samples. Reducing the expression of PLEKHO1 by specific siRNAs significantly inhibited cell viability and cell migration, and facilitated apoptosis in 786-0 and Caki-1 cells. In addition, knocking-down the expression of PLEKHO1 greatly impeded xenografted tumor's growth in nude mice. Finally, signalling pathways exploring by microarray analysis indicated that PLEKHO1-knockdown decreased the expressions of TAZ and CTGF, which are two key factors in HIPPO signalling pathway, and simultaneously reduced the expressions of p-JNK and c-Jun, which are two key factors in the JNK signalling pathway, in 786-O cells. CONCLUSIONS: These results suggest that the aberrant expression of PLEKHO1 in RCC may contribute to tumor growth, and therefore the potential of PLEKHO1 as a therapeutic target needs to be further studied. Source of Funding: National Natural Science Foundation of China (grant nos. 81372766 and 81572532), and the Liaoning “Climbing” scholarship. Shenyang, China, People's Republic of© 2019 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 201Issue Supplement 4April 2019Page: e207-e207 Advertisement Copyright & Permissions© 2019 by American Urological Association Education and Research, Inc.MetricsAuthor Information Yu Zeng* More articles by this author Zi Yu More articles by this author Shui Fu More articles by this author Gejun Zhang More articles by this author Chengcheng Lv More articles by this author Qingzhuo Dong More articles by this author Chuize Kong More articles by this author Cheng Fu More articles by this author Expand All Advertisement PDF downloadLoading ...