Read-through circular RNA rt-circ-HS promotes hypoxia inducible factor 1α expression and renal carcinoma cell proliferation, migration and invasiveness.
- Author:
Yun Yi XU
1
;
Zheng Zheng SU
1
;
Lin Mao ZHENG
1
;
Meng Ni ZHANG
1
;
Jun Ya TAN
1
;
Ya Lan YANG
1
;
Meng Xin ZHANG
1
;
Miao XU
1
;
Ni CHEN
1
;
Xue Qin CHEN
1
;
Qiao ZHOU
1
Author Information
1. Department of Pathology, West China Hospital, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- Keywords:
Hypoxia inducible factor 1α;
Read-through circular RNA HIF1α-SNAPC1;
Renal cell carcinoma;
Small nuclear RNA activating complex polypeptide 1;
microRNA 539
- MeSH:
Humans;
Carcinoma, Renal Cell/pathology*;
Cell Proliferation;
Hypoxia;
Kidney Neoplasms;
MicroRNAs/genetics*;
Neoplasm Invasiveness/genetics*;
RNA, Circular/metabolism*;
RNA, Small Interfering;
Hypoxia-Inducible Factor 1, alpha Subunit/genetics*
- From:
Journal of Peking University(Health Sciences)
2023;55(2):217-227
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To identify and characterize read-through RNAs and read-through circular RNAs (rt-circ-HS) derived from transcriptional read-through hypoxia inducible factor 1α (HIF1α) and small nuclear RNA activating complex polypeptide 1 (SNAPC1) the two adjacent genes located on chromosome 14q23, in renal carcinoma cells and renal carcinoma tissues, and to study the effects of rt-circ-HS on biological behavior of renal carcinoma cells and on regulation of HIF1α.
METHODS:Reverse transcription-polymerase chain reaction (RT-PCR) and Sanger sequencing were used to examine expression of read-through RNAs HIF1α-SNAPC1 and rt-circ-HS in different tumor cells. Tissue microarrays of 437 different types of renal cell carcinoma (RCC) were constructed, and chromogenic in situ hybridization (ISH) was used to investigate expression of rt-circ-HS in different RCC types. Small interference RNA (siRNA) and artificial overexpression plasmids were designed to examine the effects of rt-circ-HS on 786-O and A498 renal carcinoma cell proliferation, migration and invasiveness by cell counting kit 8 (CCK8), EdU incorporation and Transwell cell migration and invasion assays. RT-PCR and Western blot were used to exa-mine expression of HIF1α and SNAPC1 RNA and proteins after interference of rt-circ-HS with siRNA, respectively. The binding of rt-circ-HS with microRNA 539 (miR-539), and miR-539 with HIF1α 3' untranslated region (3' UTR), and the effects of these interactions were investigated by dual luciferase reporter gene assays.
RESULTS:We discovered a novel 1 144 nt rt-circ-HS, which was derived from read-through RNA HIF1α-SNAPC1 and consisted of HIF1α exon 2-6 and SNAPC1 exon 2-4. Expression of rt-circ-HS was significantly upregulated in 786-O renal carcinoma cells. ISH showed that the overall positive expression rate of rt-circ-HS in RCC tissue samples was 67.5% (295/437), and the expression was different in different types of RCCs. Mechanistically, rt-circ-HS promoted renal carcinoma cell proliferation, migration and invasiveness by functioning as a competitive endogenous inhibitor of miR-539, which we found to be a potent post-transcriptional suppressor of HIF1α, thus promoting expression of HIF1α.
CONCLUSION:The novel rt-circ-HS is highly expressed in different types of RCCs and acts as a competitive endogenous inhibitor of miR-539 to promote expression of its parental gene HIF1α and thus the proliferation, migration and invasion of renal cancer cells.
