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Hypoxia leads to significant changes in alternative splicing and elevated expression of CLK splice factor kinases in PC3 prostate cancer cells
- Source :
- BMC Cancer, 18:355. BioMed Central Ltd, BMC Cancer, BMC Cancer, Vol 18, Iss 1, Pp 1-11 (2018)
- Publication Year :
- 2018
- Publisher :
- BioMed Central Ltd, 2018.
-
Abstract
- Background Mounting evidence suggests that one of the ways that cells adapt to hypoxia is through alternative splicing. The aim of this study was firstly to examine the effect of hypoxia on the alternative splicing of cancer associated genes using the prostate cancer cell line PC3 as a model. Secondly, the effect of hypoxia on the expression of several regulators of splicing was examined. Methods PC3 cells were grown in 1% oxygen in a hypoxic chamber for 48 h, RNA extracted and sent for high throughput PCR analysis at the RNomics platform at the University of Sherbrooke, Canada. Genes whose exon inclusion rate PSI (ψ) changed significantly were identified, and their altered exon inclusion rates verified by RT-PCR in three cell lines. The expression of splice factors and splice factor kinases in response to hypoxia was examined by qPCR and western blotting. The splice factor kinase CLK1 was inhibited with the benzothiazole TG003. Results In PC3 cells the exon inclusion rate PSI (ψ) was seen to change by > 25% in 12 cancer-associated genes; MBP, APAF1, PUF60, SYNE2, CDC42BPA, FGFR10P, BTN2A2, UTRN, RAP1GDS1, PTPN13, TTC23 and CASP9 (caspase 9). The expression of the splice factors SRSF1, SRSF2, SRSF3, SAM68, HuR, hnRNPA1, and of the splice factor kinases SRPK1 and CLK1 increased significantly in hypoxia. We also observed that the splice factor kinase CLK3, but not CLK2 and CLK4, was also induced in hypoxic DU145 prostate, HT29 colon and MCF7 breast cancer cell lines. Lastly, we show that the inhibition of CLK1 in PC3 cells with the benzothiazole TG003 increased expression of the anti-apoptotic isoform caspase 9b. Conclusions Significant changes in alternative splicing of cancer associated genes occur in prostate cancer cells in hypoxic conditions. The expression of several splice factors and splice factor kinases increases during hypoxia, in particular the Cdc-like splice factor kinases CLK1 and CLK3. We suggest that in hypoxia the elevated expression of these regulators of splicing helps cells adapt through alternative splicing of key cancer-associated genes. We suggest that the CLK splice factor kinases could be targeted in cancers in which hypoxia contributes to resistance to therapy. Electronic supplementary material The online version of this article (10.1186/s12885-018-4227-7) contains supplementary material, which is available to authorized users.
- Subjects :
- 0301 basic medicine
Male
Cancer Research
PROTEINS
Apoptosis
CLK1
CARCINOMA-CELLS
Splice factors
Biology
SRPK1
Protein Serine-Threonine Kinases
CLK3
lcsh:RC254-282
03 medical and health sciences
Exon
0302 clinical medicine
DU145
Cell Line, Tumor
MULTIPLE
BINDING
Genetics
Humans
BREAST-CANCER
splice
TG003
Promoter Regions, Genetic
Hypoxia
GENE-EXPRESSION
Prostate cancer
Kinase
Splice factor kinases
Alternative splicing
Prostatic Neoplasms
RNA-Binding Proteins
Protein-Tyrosine Kinases
lcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens
Cell biology
Gene Expression Regulation, Neoplastic
030104 developmental biology
TARGET
Oncology
030220 oncology & carcinogenesis
Multigene Family
RNA splicing
INHIBITORS
RESISTANCE
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 14712407
- Volume :
- 18
- Database :
- OpenAIRE
- Journal :
- BMC Cancer
- Accession number :
- edsair.doi.dedup.....440d705fd3478fa8c6bcd801db622e31