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Effect of RNF8 Regulation of Wnt/β-catenin Signalling Pathway on Proliferation Invasion Migration and Epithelial Mesenchymal Transition of Renal Clear Cell Carcinoma Cells |
ZHANG Yali, ZHANG Xiaoyan, ZHAO Junlin |
Peking University First Hospital Taiyuan Hospital / Taiyuan Central Hospital, Shanxi Taiyuan 030000, China |
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Abstract Objective: To explore the effects of RNF8 on the proliferation, invasion, migration, EMT of CCRCC cells, as well as its regulatory mechanism on the Wnt/β-catenin signaling pathway. Methods: Thirty specimens of cancer and paracancerous tissues from CCRCC patients who attended and underwent pathological examination at Taiyuan Central Hospital from 2018 to 2023 were collected; RNF8 mRNA expression levels in paracancerous tissues, cancerous tissues, and human CCRCC cancer cells 786-O were detected by qRT-PCR; human CCRCC cell line 786-O cells were randomly divided into the NC group (untransfected with plasmidThe human CCRCC cell line 786-O cells were randomly divided into NC group (not transfected with plasmid), si-NC group (transfected with RNF8 negative control), si-RNF8 group (transfected with si-RNF8), and si-RNF8+Wnt activator LiCl (si-RNF8+LiCl) group; the proliferation of cells was detected by using the CCK-8 assay and the clone formation assay; the number of migrating and invasive cells in the groups was detected by using the Transwell assay; and the number of cells that had migrated and invaded in the groups was detected by using theWestern blot assay was used to detect the protein expression levels of Wnt/β-catenin signalling pathway and EMT in the cells. Results: The expression level of RNF8 mRNA in CCRCC cancer tissue (1.47±0.12) was higher than that in adjacent tissues (0.62±0.05) (t=35.813, P<0.001). Compared with the si-NC group, the expression levels of RNF8 mRNA, Wnt1, β-catenin, Vimentin, and N-cadherin, cell survival rate, clone formation number, and the numbers of cells undergoing migration and invasion in the si-RNF8 group were greatly lower (P<0.05), while the expression level of E-cadherin higher (P<0.05). The addition of Wnt activator LiCl reversed the inhibitory effect of silencing RNF8 expression on proliferation, invasion, migration, and EMT of 786-O cells (P<0.05). Conclusion: RNF8 is highly expressed in CCRCC cancer tissues, and knocking down RNF8 can inhibit the proliferation, invasion, migration, and EMT of CCRCC cancer cells, which may be related to the inhibition of the Wnt/β - catenin signaling pathway.
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