ZNF677 suppresses renal cell carcinoma progression through N6‐methyladenosine and transcriptional repression of CDKN3

Abstract

Background Studies on biological functions of N6‐methyladenosine (m 6 A) modification in mRNA have sprung up in recent years. Previous studies have reported m 6 A can determine mRNA fate and play a pivotal role in tumour development and progression. The zinc finger protein 677 (ZNF677) belongs to the zinc finger protein family and possesses transcription factor activity by binding sequence‐specific DNA. Methods The expression of ZNF677 and its clinicopathological impact were evaluated in renal cell carcinoma (RCC) patients. The m 6 A level of ZNF677 was determined by m 6 A methylated RNA immunoprecipitation‐sequencing (MeRIP‐seq) and MeRIP‐qPCR in RCC tissues and adjacent normal tissues. RNA immunoprecipitation‐qPCR (RIP‐qPCR) and luciferase assays were performed to identify the targeted effect of IGF2BP2 and YTHDF1 on ZNF677. RCC cells and subcutaneous models uncovered the role of ZNF677 methylated by CRISPR/dCas13b‐METTL3 in tumour growth. ZNF677‐binding sites in the CDKN3 promoter were investigated by chromatin immunoprecipitation (ChIP) and luciferase assays. Results ZNF677 is frequently downregulated in RCC tissues and its low expression is associated with unfavourable prognosis and decreased m 6 A modification level. Further, we find the m 6 A‐modified coding sequence (CDS) of ZNF677 positively regulates its translation and mRNA stability via binding with YTHDF1 and IGF2BP2, respectively. Targeted specific methylation of ZNF677 m 6 A by CRISPR/dCas13b‐METLL3 system can significantly increase the m 6 A and expression level of ZNF677, and dramatically inhibit cell proliferation and induce cell apoptosis of RCC cells. In addition, ZNF677 exerted its tumour suppressor functions in RCC cells through transcriptional repression of CDKN3 via binding to its promoter. In vitro and clinical data confirm the negative roles of ZNF677/CDKN3 in tumour growth and progression of RCC. Conclusion ZNF677 functions as a tumour suppressor and is frequently silenced via m 6 A modification in RCC, which may highlight m 6 A methylation‐based approach for RCC diagnosis and therapy.

Oncofetal protein IGF2BPs in human cancer: functions, mechanisms and therapeutic potential

“…PCAT6 in turn intensifies the stability effects of IGF2BP2 on IGF1R mRNA, further promoting prostate cancer growth and bone metastasis [ 47 ]. ZNF677, which plays a tumor suppressor role in renal cell carcinoma, can be stabilized by METLL3/IGF2BP2-regulated m 6 A modification at the post-transcription level [ 48 ]. IGF2BP2 stabilizes MSX1 and JARID2 transcripts in an m 6 A-dependent manner, regulating cell migration and survival in colorectal cancer [ 49 ].…”

Human pan-cancer analysis of the predictive biomarker for the CDKN3

“…It was shown that miR-127-3p promotes the proliferation and metastasis of renal cell carcinoma through CDKN 25 . And ZNF677 was also shown to inhibit the progression of renal cell carcinoma through the transcription of N6-methyladenosine and CDKN 29 . CDKN3 was moreover demonstrated to be an independent prognostic factor contributing to the progression of nasopharyngeal carcinoma to advanced stages 30 .…”

“…So, is there an association between CDKN3 and methylation or not? It has been demonstrated that ZNF677 inhibits renal cell carcinoma progression through the transcription of N6-methyladenosine and CDKN3 29 . It has also been shown that modulation of neuroblastoma cell proliferation can alter the methylation of the promoter region of the CDKN3 gene 33 .…”

ZNF582 overexpression restrains the progression of clear cell renal cell carcinoma by enhancing the binding of TJP2 and ERK2 and inhibiting ERK2 phosphorylation

“…Due to their multiple functions, some zinc finger proteins are powerful regulators in the development of tumors, including renal cancer. For example, ZNF677 inhibits RCC progression through N6-methyladenosine and transcriptional repression of CDKN3 [ 45 ]. Therefore, in addition to the regulatory effect of ZNF582 on the growth and metastasis of ccRCC in this study, more studies are also needed to explore the regulatory effects of ZNF582 on other pathways, such as cell metabolism, autophagy, immune response, stem cell maintenance and differentiation.…”