Downregulation of lncRNA ZNF582-AS1 due to DNA hypermethylation promotes clear cell renal cell carcinoma growth and metastasis by regulating the N(6)-methyladenosine modification of MT-RNR1.

Abstract

BACKGROUND:

Emerging evidence confirms that lncRNAs (long non-coding RNAs) are potential biomarkers that play vital roles in tumors. ZNF582-AS1 is a novel lncRNA that serves as a potential prognostic marker of cancers. However, the specific clinical significance and molecular mechanism of ZNF582-AS1 in ccRCC (clear cell renal cell carcinoma) are unclear.

METHODS:

Expression level and clinical significance of ZNF582-AS1 were determined by TCGA-KIRC data and qRT-PCR results of 62 ccRCCs. DNA methylation status of ZNF582-AS1 promoter was examined by MSP, MassARRAY methylation and demethylation analysis. Gain-of-function experiments were conducted to investigate the biological roles of ZNF582-AS1 in the phenotype of ccRCC. The subcellular localization of ZNF582-AS1 was detected by RNA FISH. iTRAQ, RNA pull-down and RIP-qRT-PCR were used to identify the downstream targets of ZNF582-AS1. rRNA MeRIP-seq and MeRIP-qRT-PCR were utilized to examine the N(6)-methyladenosine modification status. Western blot and immunohistochemistry assays were used to determine the protein expression level.

RESULTS:

ZNF582-AS1 was downregulated in ccRCC, and decreased ZNF582-AS1 expression was significantly correlated with advanced tumor stage, higher pathological stage, distant metastasis and poor prognosis. Decreased ZNF582-AS1 expression was caused by DNA methylation at the CpG islands within its promoter. ZNF582-AS1 overexpression inhibited cell proliferative, migratory and invasive ability, and increased cell apoptotic rate in vitro and in vivo. Mechanistically, we found that ZNF582-AS1 overexpression suppressed the N(6)-methyladenosine modification of MT-RNR1 by reducing rRNA adenine N(6)-methyltransferase A8K0B9 protein level, resulting in the decrease of MT-RNR1 expression, followed by the inhibition of MT-CO2 protein expression. Furthermore, MT-RNR1 overexpression reversed the decreased MT-CO2 expression and phenotype inhibition of ccRCC induced by increased ZNF582-AS1 expression.

CONCLUSIONS:

This study demonstrates for the first time that ZNF582-AS1 functions as a tumor suppressor gene in ccRCC and ZNF582-AS1 may serve as a potential biomarker and therapeutic target of ccRCC.

• Authors: Wuping Yang, Kenan Zhang, Lei Li, Yawei Xu, Kaifang Ma, Haibiao Xie, Jingcheng Zhou, Lin Cai, Yanqing Gong, Kan Gong
• Journal: Journal of experimental & clinical cancer research : CR (J Exp Clin Cancer Res) Vol. 40 Issue 1 Pg. 92 (Mar 10 2021) ISSN: 1756-9966 [Electronic] England
• PMID: 33691743 (Publication Type: Journal Article, Retracted Publication)

Chemical References

• Kruppel-Like Transcription Factors
• RNA, Antisense
• RNA, Long Noncoding
• RNA, Ribosomal
• RNA, ribosomal, 12S
• ZNF582 protein, human
• N-methyladenosine
• Adenosine

Topics

• Adenosine (analogs & derivatives, metabolism)
• Animals
• Carcinoma, Renal Cell (genetics, metabolism, pathology)
• Cell Line, Tumor
• DNA Methylation
• Disease Models, Animal
• Down-Regulation
• Humans
• Kidney Neoplasms (genetics, metabolism, pathology)
• Kruppel-Like Transcription Factors (genetics, metabolism)
• Mice
• Mice, Nude
• Middle Aged
• Neoplasm Metastasis
• RNA, Antisense (genetics, metabolism)
• RNA, Long Noncoding (genetics, metabolism)
• RNA, Ribosomal (genetics, metabolism)
• Transfection