Abstract
BackgroundUCA1 is frequently upregulated in a variety of cancers, including CRC, and it can play an oncogenic role by various mechanisms. However, how UCA1 is regulated in cancer is largely unknown. In this study, we aimed to determine whether RNA methylation at N6-methyladenosine (m6A) can impact UCA1 expression in colorectal cancer (CRC).MethodsqRT-PCR was performed to detect the level of UCA1 and IGF2BP2 in CRC samples. CRISPR/Cas9 was employed to knockout (KO) UCA1, METTL3 and WTAP in DLD-1 and HCT-116 cells, while rescue experiments were carried out to re-express METTL3 and WTAP in KO cells. Immunoprecipitation using m6A antibody was performed to determine the m6A modification of UCA1. In vivo pulldown assays using S1m tagging combined with site-direct mutagenesis was carried out to confirm the recognition of m6A-modified UCA1 by IGF2BP2. Cell viability was measured by MTT and colony formation assays. The expression of UCA1 and IGF2BP2 in TCGA CRC database was obtained from GEPIA (http://gepia.cancer-pku.cn).ResultsOur results revealed that IGF2BP2 serves as a reader for m6A modified UCA1 and that adenosine at 1038 of UCA1 is critical to the recognition by IGF2BP2. Importantly, we showed that m6A writers, METTL3 and WTAP positively regulate UCA1 expression. Mechanically, IGF2BP2 increases the stability of m6A-modified UCA1. Clinically, IGF2BP2 is upregulated in CRC tissues compared with normal tissues.ConclusionThese results suggest that m6A modification is an important factor contributing to upregulation of UCA1 in CRC tissues.
Highlights
Colorectal cancer (CRC) is the third most commonly diagnosed cancer, and is the second leading cause of cancer-related death worldwide [1]
Upregulation of Urothelial carcinoma associated 1 (UCA1) in colorectal cancer (CRC) tissues To determine the expression of UCA1 in CRC, we first analyzed CRC data from TCGA
We found that the level of UCA1 was significantly higher higher in CRC (n = 599) than in normal tissues (Fig. 1A)
Summary
Colorectal cancer (CRC) is the third most commonly diagnosed cancer, and is the second leading cause of cancer-related death worldwide [1]. Despite declines in incidence over the past decade owing to adoption of effective screening programs, approximately 50% of patients. He et al Cancer Cell International (2021) 21:616 show metastasis at the time of diagnosis [3]. Evidence accumulated over the past decade indicates that the lncRNAs are frequently dysregulated in cancer and they can play key roles in tumorigenesis. Urothelial carcinoma associated 1 (UCA1) is one of the most well-known lncRNAs and is highly expressed in CRC and tightly associated with the development and progression of CRC [12,13,14]. We aimed to determine whether RNA methylation at N6-methyladenosine (m6A) can impact UCA1 expression in colorectal cancer (CRC)
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