Abstract
Pseudogenes have long been considered as nonfunctional genomic sequences. Recent studies have shown that they can potentially regulate the expression of protein-coding genes and are dysregulated in diseases including cancer. However, the potential roles of pseudogenes in ovarian cancer have not been well studied. Here we characterized the pseudogene expression profile in HGSOC (high-grade serous ovarian carcinoma) by microarray. We identified 577 dysregulated pseudogenes and most of them were up-regulated (538 of 577). HMGA1P6 (High mobility group AT-hook 1 pseudogene 6) was one of the overexpressed pseudogenes and its expression was inversely correlated with patient survival. Mechanistically, HMGA1P6 promoted ovarian cancer cell malignancy by acting as a ceRNA (competitive endogenous RNA) that led to enhanced HMGA1 and HMGA2 expression. Importantly, HMGA1P6 was transcriptionally activated by oncogene MYC in ovarian cancer. Our findings reveal that MYC may contribute to oncogenesis through transcriptional regulation of pseudogene HMGA1P6 in ovarian cancer.
Highlights
Ovarian cancer is the most lethal gynecologic cancer.Globally, there are 239,000 new cases (3.6% of all cancer cases) and 152,000 deaths reported annually[1]
We further demonstrated that HMGA1P6 was one of highly expressed pseudogenes in HGSOC which promoted ovarian cancer aggressiveness through modulating HMGA1/2
Hierarchical cluster and correlation matrix heat map analysis showed that HGSOC exhibited specific pseudogene expression pattern compared to normal tissues (Fig. 1a, b)
Summary
RNA isolation and qPCR Total RNA was extracted from cells or fresh tissues with Cell Total RNA Isolation Kit (Foregene, Chengdu, China) following manufacturer’s instructions. RIP assay RIP (RNA Immunoprecipitation) assay was performed using EZ-Magna RIP RNA-Binding Protein Immunoprecipitation Kit (Merck KGaA, Darmstadt, Germany) following manufacturer’s instructions. Cells were lysed with RIP lysis buffer and incubated with magnetic beads coated with anti-Ago[2] antibody (Merck) and IgG (Millipore) was used as a negative control. QRT-PCR was performed to detect the enrichment of HMGA1P6 and miRNAs. RNA pull-down assay Biotin‐labeled HMGA1P6 transcripts was obtained using in vitro transcription with T7 RNA polymerase from the TranscriptAid T7 High Yield Transcription Kit (Thermo Fisher Scientific). RNA pull-down was performed using the Magnetic RNA-Protein Pull-Down kit (Thermo Fisher Scientific, USA) according to manufacturer’s protocol. Significance was considered as: *p < 0.05, **p < 0.01
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