Small activating RNA induced expression of VHL gene in renal cell carcinoma.

Abstract

Recent studies have reported that chemically synthesized double-stranded RNAs (dsRNAs), also known as small activating RNA (saRNAs), can specifically induce gene expression by targeting promoter sequences by a mechanism termed RNA activation (RNAa). In the present study, we designed 4 candidate saRNAs targeting the Von Hippel-Lindau (VHL) gene promoter. Among these saRNAs, dsVHL-821 significantly inhibited cell growth by up-regulating VHL at both the mRNA and protein levels in renal cell carcinoma 769-P cells. Functional analysis showed that dsVHL-821 induced apoptosis by increasing p53, decreasing Bcl-xL, activating caspase 3/7 and poly-ADP-ribose polymerase in a dose-dependent manner. Chromatin immunoprecipitation analysis revealed that dsVHL-821 increased the enrichment of Ago2 and RNA polymerase II at the dsVHL-821 target site. In addition, Ago2 depletion significantly suppressed dsVHL-821-induced up-regulation of VHL gene expression and related effects. Single transfection of dsVHL-821 caused long-lasting (14 days) VHL up-regulation. Furthermore, the activation of VHL by dsVHL-821 was accompanied by an increase in dimethylation of histone 3 at lysine 4 (H3K4me2) and acetylation of histone 4 (H4ac) and a decrease in dimethylation of histone 3 at lysine 9 (H3K9me2) and lysine 27 (H3K27me2) in the dsVHL-821 target region. Taken together, these results demonstrate that dsVHL-821, a novel saRNA for VHL, induces the expression of the VHL gene by epigenetic changes, leading to inhibition of cell growth and induction of apoptosis, and suggest that targeted activation of VHL by dsVHL-821 may be explored as a novel treatment of renal cell carcinoma.

• Authors: Moo Rim Kang, Ki Hwan Park, Chang Woo Lee, Myeong Youl Lee, Sang-Bae Han, Long-Cheng Li, Jong Soon Kang
• Journal: The international journal of biochemistry & cell biology (Int J Biochem Cell Biol) Vol. 97 Pg. 36-42 (04 2018) ISSN: 1878-5875 [Electronic] Netherlands
• PMID: 29425832 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2018 Elsevier Ltd. All rights reserved.

Chemical References

• RNA, Double-Stranded
• Von Hippel-Lindau Tumor Suppressor Protein
• VHL protein, human

Topics

• Apoptosis (drug effects)
• Carcinoma, Renal Cell (drug therapy, genetics, metabolism, pathology)
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Epigenesis, Genetic (drug effects)
• Gene Expression Regulation, Neoplastic (drug effects)
• Humans
• Kidney Neoplasms (drug therapy, genetics, metabolism, pathology)
• RNA, Double-Stranded (pharmacology)
• Von Hippel-Lindau Tumor Suppressor Protein (biosynthesis, genetics)