A peptide CORO1C-47aa encoded by the circular noncoding RNA circ-0000437 functions as a negative regulator in endometrium tumor angiogenesis

Abstract

Circular RNAs (circRNAs) are a novel class of widespread noncoding RNAs that regulate gene expression in mammals. Recent studies demonstrate that functional peptides can be encoded by short open reading frames in noncoding RNAs, including circRNAs. However, the role of circRNAs in various physiological and pathological states, such as cancer, is not well understood. In this study, through deep RNA sequencing on human endometrial cancer (EC) samples and their paired adjacent normal tissues, we uncovered that the circRNA hsa-circ-0000437 is significantly reduced in EC compared with matched paracancerous tissue. The hsa-circ-0000437 contains a short open reading frame encoding a functional peptide termed CORO1C-47aa. Overexpression of CORO1C-47aa is capable of inhibiting angiogenesis at the initiation stage by suppressing endothelial cell proliferation, migration, and differentiation through competition with transcription factor TACC3 to bind to ARNT and suppress VEGF. CORO1C-47aa directly bound to ARNT through the PAS-B domain, and blocking the association between ARNT and TACC3, which led to reduced expression of VEGF, ultimately lead to reduced angiogenesis. The antitumor effects of CORO1C-47aa on EC progression suggest that CORO1C-47aa has potential value in anticarcinoma therapies and warrants further investigation. Circular RNAs (circRNAs) are a novel class of widespread noncoding RNAs that regulate gene expression in mammals. Recent studies demonstrate that functional peptides can be encoded by short open reading frames in noncoding RNAs, including circRNAs. However, the role of circRNAs in various physiological and pathological states, such as cancer, is not well understood. In this study, through deep RNA sequencing on human endometrial cancer (EC) samples and their paired adjacent normal tissues, we uncovered that the circRNA hsa-circ-0000437 is significantly reduced in EC compared with matched paracancerous tissue. The hsa-circ-0000437 contains a short open reading frame encoding a functional peptide termed CORO1C-47aa. Overexpression of CORO1C-47aa is capable of inhibiting angiogenesis at the initiation stage by suppressing endothelial cell proliferation, migration, and differentiation through competition with transcription factor TACC3 to bind to ARNT and suppress VEGF. CORO1C-47aa directly bound to ARNT through the PAS-B domain, and blocking the association between ARNT and TACC3, which led to reduced expression of VEGF, ultimately lead to reduced angiogenesis. The antitumor effects of CORO1C-47aa on EC progression suggest that CORO1C-47aa has potential value in anticarcinoma therapies and warrants further investigation. Endometrial cancer (EC) is the fifth most common malignancy in women worldwide accounting for 1% to 2% of all deaths from cancer (1Lee T.Y. Martinez-Outschoorn U.E. Schilder R.J. Kim C.H. Richard S.D. Rosenblum N.G. Johnson J.M. Metformin as a therapeutic target in endometrial cancers.Front. Oncol. 2018; 8: 341Crossref PubMed Scopus (40) Google Scholar, 2Ferlay J. Soerjomataram I. Dikshit R. Eser S. Mathers C. Rebelo M. Parkin D.M. Forman D. Bray F. Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012.Int. J. Cancer. 2015; 136: E359-E386Crossref PubMed Scopus (20455) Google Scholar). The incidence of EC has steadily increased and is noted to be much higher in developed countries than in developing countries in the past 10 years, and nearly 11.7% of the cases occur in China (3He Y. Tao X. Huang F. Jia N. Du Y. Yu J. Feng W. Clinicopathologic features of endometrial cancer in Chinese patients younger than 50 years with a family history of cancer.Medicine. 2018; 97e12968Crossref PubMed Scopus (6) Google Scholar). The risk factors for developing EC include age, high socioeconomic status, and factors related to an excess of estrogen exposure, diabetes, or hypertension, but relatively little is known about inherited risk factors for this disease (4Potischman N. Hoover R.N. Brinton L.A. Siiteri P. Dorgan J.F. Swanson C.A. Berman M.L. Mortel R. Twiggs L.B. Barrett R.J. Wilbanks G.D. Persky V. Lurain J.R. Case-control study of endogenous steroid hormones and endometrial cancer.J. Natl. Cancer Inst. 1996; 88: 1127-1135Crossref PubMed Scopus (261) Google Scholar). Thus, there is a need for a deeper understanding of the molecular pathways involved in the occurrence and development of EC. Noncoding RNAs (ncRNAs) represent the majority of transcripts in a cell and are known to be involved in the development of EC (5Li N. Zheng J. Li H. Deng J. Hu M. Wu H. Li W. Li F. Lan X. Lu J. Zhou Y. Identification of chimeric TSNAX-DISC1 resulting from intergenic splicing in endometrial carcinoma through high-throughput RNA sequencing.Carcinogenesis. 2014; 35: 2687-2697Crossref PubMed Scopus (32) Google Scholar, 6Li W. Li H. Zhang L. Hu M. Li F. Deng J. An M. Wu S. Ma R. Lu J. Zhou Y. Long non-coding RNA LINC00672 contributes to p53 protein-mediated gene suppression and promotes endometrial cancer chemosensitivity.J. Biol. Chem. 2017; 292: 5801-5813Abstract Full Text Full Text PDF PubMed Scopus (44) Google Scholar). Circular RNAs (circRNAs) are a novel class of widespread ncRNAs that regulate gene expression in mammals. They are closed RNA transcripts generated by back-splicing of a single pre-mRNA. About 85% of circRNAs are aligned in sense orientation to known protein-coding genes and span 1 to 5 exons of the housed protein-coding genes. The expression of circRNAs is often highly conserved across species (7Servick K. 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However, the function of most circRNAs is the development of sequencing and studies that functional peptides can be encoded by short open reading frames in ncRNAs R.J. U. M. of open reading frames by 2014; PubMed Scopus Google Scholar, J.A. for functional peptides encoded by short open reading 2014; PubMed Scopus Google Scholar). to the of some circRNAs can be through the by Y. X. M. X. Wu P. Zhang Y. Y. Y. Wang Y. X. Wang of RNAs by 2017; PubMed Scopus Google Scholar). For and to be I. F. M. F. A. A. M. P. N. I. is a RNA that can be and in 2017; Full Text Full Text PDF PubMed Scopus Google Scholar). by an a novel Y. X. Zhang M. S. C. F. Huang N. X. K. Zhou H. Huang S. B. Zhang N. role of RNA in Natl. Cancer Inst. 2018; Scopus Google Scholar). generated from the contains a encoding a peptide in human M. K. X. Y. S. P. Liu H. J. F. Zhou H. X. Huang N. Liu J. K. K. et peptide encoded by of in 2018; PubMed Scopus Google Scholar). In this we generated deep RNA sequencing from EC samples and their paired adjacent normal and circRNA that the human circRNA hsa-circ-0000437 is in cancer and paracancerous contains a encoding a peptide that is termed CORO1C-47aa. further the functional role of this peptide in EC development and that CORO1C-47aa with TACC3 to with the PAS-B of to suppression of and of angiogenesis in EC. the circRNAs that are involved in the of we on the RNA from human EC samples and their paired adjacent The of circRNAs from all samples from to with the in of compared with their matched adjacent normal About of the circRNAs is known circRNAs in the circRNA P. P. N. for 2014; PubMed Scopus Google of all circRNAs from protein-coding exons and aligned with intergenic The of circRNAs is to and there between the and the normal we compared the expression of circRNAs in EC and their matched adjacent normal of all circRNAs in of The further hsa-circ-0000437 as the circRNA that expression in than matched normal and circRNA as the most significantly circRNA in normal is in EC compared with their matched adjacent tissues, we that the of hsa-circ-0000437 to the of in a of paired EC samples and matched significantly in EC compared with their matched adjacent However, the circRNA be by not from the of the and exons of gene is in we of for hsa-circ-0000437 a that the and a that the that the the and of EC as and as with the on in to the RNAs, the circRNA resistant to the of a highly to that not on circRNAs and by the R.J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, of the Biol. PubMed Scopus Google thus the of sequencing of the of hsa-circ-0000437 the of that matched the in we the of transcripts from the hsa-circ-0000437 with a transcription and at for RNA little in with for the of the from the with to the of we the and of EC The of the and by which a of the and The of by that the in the RNA in to the of In with the in the of EC further a in a circRNA expression which by with splicing and However, the that the not is known that circRNAs can as miRNA to regulate gene expression in cell H. S. Li W. Yu P. The RNA and transcription and in 2015; PubMed Scopus Google Scholar). EC development through the we RNA an a molecular sponge of I. F. M. F. A. A. M. P. N. I. is a RNA that can be and in 2017; Full Text Full Text PDF PubMed Scopus Google in a of However, not in that regulate the occurrence and development of EC through some circRNAs to functional we the of and a with the potential to a peptide in from the in the of the gene the to a in the of the which we termed as CORO1C-47aa and and with the human that CORO1C-47aa peptide in this is we to The associated with but not and of and a circRNA with in by that but not in the a of a but of in the that be Recent studies that the of circRNAs on the of an a regulatory that is for Zhou K. J.A. is for differentiation in PubMed Scopus Google Scholar). matched in for human circRNAs. the of we a of the of in this is we the or between the and genes of a and the of the of the of the that with the the compared with the In or of the significantly reduced the of by that the of is the protein-coding of we the a that to has splicing and the has sequencing the is of the a can be the of a in and of the increased the expression of by in and EC which not by a the the CORO1C-47aa can be in or by we generated an the which the expression of endogenous and we an of the of CORO1C-47aa can the initiation of an expression in which the encoding to and a to the of CORO1C-47aa. of CORO1C-47aa in with the but not in with the by CORO1C-47aa is in EC compared with normal EC we generated cell with stable of or a with the not the expression of of led to of CORO1C-47aa we that CORO1C-47aa the of and EC we that target the of reduced expression and the CORO1C-47aa peptide with on the expression of the gene and The expression of the peptide by to target and stable of not CORO1C-47aa expression and that the the peptide CORO1C-47aa of the of CORO1C-47aa on cell proliferation, we on the and that CORO1C-47aa. The from the in the cell of or compared with the we the of CORO1C-47aa on in The of EC the significantly than of their or that CORO1C-47aa but not the circRNA the The increased from 5 in to in CORO1C-47aa The increased from in to in CORO1C-47aa that the of EC in but not in we that CORO1C-47aa regulate development the of through the by CORO1C-47aa to the of we the expression of a of H. Du W. Zhang L. Y. K. cancer cell in by angiogenesis and blocking PubMed Scopus Google by with the and the expression of significantly reduced in the CORO1C-47aa In an endothelial to CORO1C-47aa endothelial on The that CORO1C-47aa significantly the of to compared with the from endothelial cell proliferation, migration, and we endothelial in from and with CORO1C-47aa we The endothelial cell significantly in the from cell CORO1C-47aa to the of CORO1C-47aa expression on cell migration, we and in in and the and that the of significantly in the from with CORO1C-47aa compared with with an or an we the of CORO1C-47aa on the endothelial cell In in from that the the a which is not to cell S. in endothelial 2017; PubMed Scopus Google Scholar). In the of with to of the of by in the and the of significantly with in from further the that CORO1C-47aa can suppress to in in the in mice with is significantly in the CORO1C-47aa and the of CORO1C-47aa in an in of angiogenesis in which a endothelial as a of the of the that with and P. S. C. F. R. M. M. of endothelial factor to the of 2017; PubMed Scopus Google Scholar). in and CORO1C-47aa a of that CORO1C-47aa is capable of inhibiting angiogenesis at the initiation stage by suppressing endothelial cell proliferation, migration, and the potential molecular the of we an in and to the of CORO1C-47aa by and in and and with in cell the the between and endogenous ARNT in cell The an in with ARNT and CORO1C-47aa CORO1C-47aa further the between CORO1C-47aa and ARNT is the for the and which as for and R.J. M.L. A. The family of J. Biol. PubMed Scopus Google Scholar). the functional that the we ARNT with of or the functional of ARNT 1 to contains functional to contains the PAS-B to contains the 1 to contains the at as the ARNT but of the and PAS-B the in but not the is for the with that CORO1C-47aa bound to ARNT through PAS-B has that the PAS-B of ARNT directly with the and promotes angiogenesis through the expression of the target gene new role for in by the Google Scholar, for of the are directly by ARNT Natl. U. S. A. PubMed Scopus Google Scholar). CORO1C-47aa with TACC3 to bind to ARNT and suppress we in EC an or the with or the to CORO1C-47aa. In and of but not or the with significantly reduced the between ARNT and TACC3 further the between in the and the ARNT and TACC3 we in EC ARNT and TACC3 bound to the of with the of the in which by of CORO1C-47aa the of CORO1C-47aa on we the of the to a with the The of EC with and an higher than that of EC with and an and that the is of CORO1C-47aa by the significantly reduced the by on that of the in ARNT and in the TACC3 significantly the of a for of the are directly by ARNT Natl. U. S. A. PubMed Scopus Google we the with or TACC3, and CORO1C-47aa. The that of and ARNT the transcription of and of TACC3 increased the which by CORO1C-47aa that CORO1C-47aa with TACC3 in the of CORO1C-47aa can the of in the cell the of by The that CORO1C-47aa in in and CORO1C-47aa led to higher The of in the to the of and in EC in the expression of and reduced in the the expression of ARNT not that CORO1C-47aa to ARNT and the of TACC3 on resulting in the suppression of that large of circRNAs in the human which in function (8Memczak S. Jens M. Elefsinioti A. Torti F. Krueger J. Rybak A. Maier L. Mackowiak S.D. Gregersen L.H. Munschauer M. Loewer A. Ziebold U. Landthaler M. Kocks C. Le Noble F. et al.Circular RNAs are a large class of animal RNAs with regulatory potency.Nature. 2013; 495: 333-338Crossref PubMed Scopus (4716) Google Scholar, Li Wu C.H. W. The RNA in the molecular human 2017; 8: PubMed Scopus Google Scholar, C. W. Li S. J. B. Y. D. Li Y. L. J. X. Huang S. Circular RNA an that cell by PubMed Scopus Google Scholar). 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Chem. 2015; Full Text Full Text PDF PubMed Scopus Google Scholar). study in ARNT as the common of CORO1C-47aa. by that the of TACC3 reduced in The that TACC3 the of ARNT to and promotes gene which by of that CORO1C-47aa with TACC3 to bind to the PAS-B of ARNT and suppress transcription and Thus, study an of expression is which warrants further in cell to this is a common for In we that CORO1C-47aa function as a in angiogenesis blocking the association between ARNT and TACC3 and the expression of to a in expression and of which ultimately to reduced angiogenesis a new between CORO1C-47aa and angiogenesis and the of CORO1C-47aa in by this is that CORO1C-47aa is an target in of EC. The in this study are of the RNA expression we a of paired EC and matched from the and of of human EC and matched for from the of of the patients or the The of and this studies by the of The of all the patients are in as 1 to 10 of RNA in of with and of at and by to the expression of as W. Li H. Zhang L. Hu M. Li F. Deng J. An M. Wu S. Ma R. Lu J. Zhou Y. Long non-coding RNA LINC00672 contributes to p53 protein-mediated gene suppression and promotes endometrial cancer chemosensitivity.J. Biol. Chem. 2017; 292: 5801-5813Abstract Full Text Full Text PDF PubMed Scopus (44) Google Scholar). The are in on RNAs from EC samples and their normal as (8Memczak S. Jens M. Elefsinioti A. Torti F. Krueger J. Rybak A. Maier L. Mackowiak S.D. Gregersen L.H. Munschauer M. Loewer A. Ziebold U. Landthaler M. Kocks C. Le Noble F. et al.Circular RNAs are a large class of animal RNAs with regulatory potency.Nature. 2013; 495: 333-338Crossref PubMed Scopus (4716) Google Scholar). RNAs with to the RNAs as and a by a from and on the to the The the The from of the and aligned to the to the In to the of all circRNA be by The circRNA the of than and than mice that to 5 of from the at the Chinese of the mice in with the by the of EC cell and from the of of the Chinese of and of The cell for than and in which with and at in the of to the In RNA from the of RNA with by 1% and from to RNA on the by at for by The by a at The with and 1 with at the the by and or RNA in EC RNA with in EC cell to the on with at for and with The samples with at for and with at in the a In the the with with with and at in the for 5 and with at the with in the for 10 The and by and as and a to the The to 10 of to an and at the and by or with the The peptide samples on samples an with a and The from to in to in at a of at in the with an with at a of and the most for The at at a of by the for as the with a of The for and to and The of and and of on The peptide to and a of The to for peptide and the to The with and the 10 to the the by cell and the at and and and to The are in as on and to at with and and with a the CORO1C-47aa peptide by by The ARNT TACC3 and mouse from from and The to the The of this the at 1 well in with and to with the in The as a of to on at and well of a the at for for in cell the for The resulting with a and with of an of to the The of samples in to the by human in and for a of as in with by for and with for and to as a with and a with in the of in the and in a in The with of to the in at in an at that through the with and with in generated as H. S. M. S. D. F. A. L. F. V. D. F. D. M. is a novel PubMed Scopus Google Scholar). a and a and to at for and various on of the The for angiogenesis by the of for mice with of in mice and and The the with a in The to the the Y. A. J. M. S. A. J. M. J. J. S. et and related and in for PubMed Scopus Google with the The is that the of this study are the and or are contains W. Li H. Zhang L. Hu M. Li F. Deng J. An M. Wu S. Ma R. Lu J. Zhou Y. Long non-coding RNA LINC00672 contributes to p53 protein-mediated gene suppression and promotes endometrial cancer chemosensitivity.J. Biol. Chem. 2017; 292: 5801-5813Abstract Full Text Full Text PDF PubMed Scopus (44) Google Scholar, S. Jens M. Elefsinioti A. Torti F. Krueger J. Rybak A. Maier L. Mackowiak S.D. Gregersen L.H. Munschauer M. Loewer A. Ziebold U. Landthaler M. Kocks C. Le Noble F. et al.Circular RNAs are a large class of animal RNAs with regulatory potency.Nature. 2013; 495: 333-338Crossref PubMed Scopus (4716) Google Scholar, H. S. M. S. D. F. A. L. F. V. D. F. D. M. is a novel PubMed Scopus Google Scholar). The that of with the of this all in this by the from the Natural of China F. Y. and H. L. S. L. N. X. L. Y. F. Y. and H. L. S. L. and N. L. Y. N. and X. C. L. H. and Y. Y. S. N. X. H. F. Y. and L. J. F. Y. N. X. and L. J. F. Y. and H. L. F. L. and H. L. F. Y. L. Y. and H. L. F. L. and H. L. with and with

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