The tumor suppressor gene RhoBTB1 is a novel target of miR-31 in human colon cancer

Xu,Rui-Si; Wu,Xiao-Dong; Zhang,Si-Qi; Li,Chang-Feng; Yang,Lei; Li,Dan-Dan; Zhang,Bao-Gang; Zhang,Ying; Jin,Jing-Peng; Zhang,Bin

doi:10.3892/ijo.2012.1746

The tumor suppressor gene RhoBTB1 is a novel target of miR-31 in human colon cancer

Authors:
- Rui-Si Xu
- Xiao-Dong Wu
- Si-Qi Zhang
- Chang-Feng Li
- Lei Yang
- Dan-Dan Li
- Bao-Gang Zhang
- Ying Zhang
- Jing-Peng Jin
- Bin Zhang
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Affiliations: Endoscopy Center, China-Japan Union Hospital, Jilin University, Changchun 130033, P.R. China
Published online on: December 20, 2012 https://doi.org/10.3892/ijo.2012.1746
Pages: 676-682

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Abstract

miRNAs are a class of endogenous non-coding RNA, which can regulate downstream target genes through binding to the 3'UTR of those genes. Numerous studies have indicated that abnormal expression of miRNAs is implicated in tumor development. Aberrant expression of miR-31 has been found in various cancers, including colorectal cancer. Here, we show that miR-31 is upregulated in human colon cancer tissues and cell lines, and that repression of miR-31 inhibited colon cancer cell proliferation and colony formation in soft agarose. To further elucidate the mechanism underlying the role of miR-31 in promoting colon cancer, we used online miRNA target prediction databases and found that the tumor suppressor RhoTBT1 may be a target of miR-31. Imunohistochemistry assay revealed that RhoBTB1 was significantly decreased in HT29 cells. In addition, ectopic expression of miR-31 reduced RhoBTB1 in the colon cancer cell line HT29. The results suggested that suppression of RhoBTB1 may be responsible for colon tumorigenesis, which was inhibited directly by miR-31. The results of MTT and soft agarose colony-formation assays showed that knockdown of RhoBTB1 by RNAi induced cell proliferation, and colony formation in soft agarose, which mimicked the function of miR-31. This further suggested that suppression of RhoBTB1 was responsible for colon tumorigenesis. In conclusion, we found that miR-31 acts as an oncogene in colon cancer and identified RhoBTB1 as a new target of miR-31 further study demonstrated that miR-31 contributed to the development of colon cancer at least partly by targeting RhoBTB1.

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1.	Jemal A, Siegel R, Xu J and Ward E: Cancer statistics, 2010. CA Cancer J Clin. 60:277–300. 2010. View Article : Google Scholar
2.	Colorectal Cancer Survival by Stage: National Cancer Intelligence Network. 2009
3.	Figer A, Perez-Staub N, Carola E, et al: FOLFOX in patients aged between 76 and 80 years with metastatic colorectal cancer: an exploratory cohort of the OPTIMOX1 study. Cancer. 110:2666–2671. 2007.PubMed/NCBI
4.	Becker H: Surgery of colorectal carcinoma. Praxis. 84:1371–1372. 1995.
5.	Jemal A, Siegel R, Ward E, Murray T, Xu J, Smigal C and Thun MJ: Cancer statistics, 2006. CA Cancer J Clin. 56:106–130. 2006. View Article : Google Scholar
6.	Tsai HL, Yeh YS, Yu FJ, Lu CY, Chen CF, Chen CW, Chang YT and Wang JY: Predicting factors of postoperative relapse in T2-4N0M0 colorectal cancer patients via harvesting a minimum of 12 lymph nodes. Int J Colorectal Dis. 24:177–183. 2009. View Article : Google Scholar : PubMed/NCBI
7.	Longo WE and Johnson FE: The preoperative assessment and postoperative surveillance of patients with colon and rectal cancer. Surg Clin North Am. 82:1091–1108. 2002. View Article : Google Scholar : PubMed/NCBI
8.	Baraniskin A, Birkenkamp-Demtroder K, Maghnouj A, et al: MiR-30a-5p suppresses tumor growth in colon carcinoma by targeting DTL. Carcinogenesis. 33:732–739. 2012. View Article : Google Scholar : PubMed/NCBI
9.	Friedman RC, Farh KK, Burge CB and Bartel DP: Most mammalian mRNAs are conserved targets of microRNAs. Genome Res. 19:92–105. 2009. View Article : Google Scholar : PubMed/NCBI
10.	Reinhart BJ, Slack FJ, Basson M, Pasquinelli AE, Bettinger JC, Rougvie AE, Horvitz HR and Ruvkun G: The 21-nucleotide let-7 RNA regulates developmental timing in Caenorhabditis elegans. Nature. 403:901–906. 2000. View Article : Google Scholar : PubMed/NCBI
11.	Chen CZ, Li L, Lodish HF and Bartel DP: MicroRNAs modulate hematopoietic lineage differentiation. Science. 303:83–86. 2004. View Article : Google Scholar : PubMed/NCBI
12.	Lai EC: Micro RNAs are complementary to 3′UTR sequence motifs that mediate negative post-transcriptional regulation. Nat Genet. 30:363–364. 2002.
13.	Engels BM and Hutvagner G: Principles and effects of microRNA-mediated post-transcriptional gene regulation. Oncogene. 25:6163–6169. 2006. View Article : Google Scholar : PubMed/NCBI
14.	Cheng AM, Byrom MW, Shelton J and Ford LP: Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Res. 33:1290–1297. 2005. View Article : Google Scholar : PubMed/NCBI
15.	Calin GA, Ferracin M, Cimmino A, et al: A MicroRNA signature associated with prognosis and progression in chronic lymphocytic leukemia. N Engl J Med. 353:1793–1801. 2005. View Article : Google Scholar : PubMed/NCBI
16.	Bushati N and Cohen SM: microRNA functions. Annu Rev Cell Dev Biol. 23:175–205. 2007. View Article : Google Scholar
17.	Garzon R, Calin GA and Croce CM: MicroRNAs in cancer. Annu Rev Med. 60:167–179. 2009. View Article : Google Scholar
18.	Slaby O, Svoboda M, Michalek J and Vyzula R: MicroRNAs in colorectal cancer: translation of molecular biology into clinical application. Mol Cancer. 8:1022009. View Article : Google Scholar : PubMed/NCBI
19.	Schetter AJ and Harris CC: Alterations of microRNAs contribute to colon carcinogenesis. Semin Oncol. 38:734–742. 2011. View Article : Google Scholar : PubMed/NCBI
20.	Bandres E, Cubedo E, Agirre X, et al: Identification by real-time PCR of 13 mature microRNAs differentially expressed in colorectal cancer and non-tumoral tissues. Mol Cancer. 5:292006. View Article : Google Scholar : PubMed/NCBI
21.	Slaby O, Svoboda M, Fabian P, Smerdova T, Knoflickova D, Bednarikova M, Nenutil R and Vyzula R: Altered expression of miR-21, miR-31, miR-143 and miR-145 is related to clinicopathologic features of colorectal cancer. Oncology. 72:397–402. 2007. View Article : Google Scholar : PubMed/NCBI
22.	Wong QW, Lung RW, Law PT, Lai PB, Chan KY, To KF and Wong N: MicroRNA-223 is commonly repressed in hepatocellular carcinoma and potentiates expression of Stathmin1. Gastroenterology. 135:257–269. 2008. View Article : Google Scholar : PubMed/NCBI
23.	Liu X, Sempere LF, Ouyang H, et al: MicroRNA-31 functions as an oncogenic microRNA in mouse and human lung cancer cells by repressing specific tumor suppressors. J Clin Invest. 120:1298–1309. 2010. View Article : Google Scholar : PubMed/NCBI
24.	Liu X, Chen Z, Yu J, Xia J and Zhou X: MicroRNA profiling and head and neck cancer. Comp Funct Genomics. 8375142009.PubMed/NCBI
25.	Valastyan S, Reinhardt F, Benaich N, Calogrias D, Szasz AM, Wang ZC, Brock JE, Richardson AL and Weinberg RA: A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis. Cell. 137:1032–1046. 2009. View Article : Google Scholar : PubMed/NCBI
26.	Laurila EM, Sandström S, Rantanen LM, Autio R and Kallioniemi A: Both inhibition and enhanced expression of miR-31 lead to reduced migration and invasion of pancreatic cancer cells. Genes Chromosomes Cancer. 51:557–568. 2012. View Article : Google Scholar : PubMed/NCBI
27.	Cottonham CL, Kaneko S and Xu L: miR-21 and miR-31 converge on TIAM 1 to regulate migration and invasion of colon carcinoma cells. J Biol Chem. 285:35293–35302. 2010. View Article : Google Scholar : PubMed/NCBI
28.	Wang CJ, Stratmann J, Zhou ZG and Sun XF: Suppression of microRNA-31 increases sensitivity to 5-FU at an early stage, and affects cell migration and invasion in HCT-116 colon cancer cells. BMC Cancer. 10:6162010. View Article : Google Scholar : PubMed/NCBI
29.	Hamaguchi M, Meth JL, von Klitzing C, et al: DBC2, a candidate for a tumor suppressor gene involved in breast cancer. Proc Natl Acad Sci USA. 99:13647–13652. 2002. View Article : Google Scholar
30.	Knowles MA, Aveyard JS, Taylor CF, Harnden P and Bass S: Mutation analysis of the 8p cand idate tumour suppressor genes DBC2 (RHOBT B2) and LZTS1 in bladder cancer. Cancer Lett. 225:121–130. 2005. View Article : Google Scholar : PubMed/NCBI
31.	Cho YG, Choi BJ, Kim CJ, Song JH, Zhang C, Nam SW, Lee JY and Park WS: Genetic analysis of the DBC2 gene in gastric cancer. Acta Oncol. 47:366–371. 2008. View Article : Google Scholar : PubMed/NCBI
32.	Beder LB, Gunduz M, Ouchida M, et al: Identification of a candidate tumor suppressor gene RHOBTB1 located at a novel allelic loss region 10q21 in head and neck cancer. J Cancer Res Clin Oncol. 132:19–27. 2006. View Article : Google Scholar : PubMed/NCBI
33.	Hwang HW and Mendel JT: MicroRNAs in cell proliferation, cell death, and tumorigenesis. Br J Cancer. 94:776–780. 2006. View Article : Google Scholar : PubMed/NCBI
34.	Lewis BP, Burge CB and Bartel DP: Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets. Cell. 120:15–20. 2005. View Article : Google Scholar : PubMed/NCBI
35.	Cho WC: MicroRNAs in cancer from research to therapy. Biochim Biophys Acta. 1805:209–217. 2010.PubMed/NCBI
36.	Cho WC: MicroRNAs: potential biomarkers for cancer diagnosis, prognosis and targets for therapy. Int J Biochem Cell Biol. 42:1273–1281. 2010. View Article : Google Scholar : PubMed/NCBI
37.	Liu CJ, Tsai MM, Hung PS, et al: miR-31 ablates expression of the HIF regulatory factor FIH to activate the HIF pathway in head and neck carcinoma. Cancer Res. 70:1635–1644. 2010. View Article : Google Scholar : PubMed/NCBI
38.	Sahai E and Marshall CJ: RHO-GTPases and cancer. Nat Rev Cancer. 2:133–142. 2002. View Article : Google Scholar
39.	Gómez del Pulgar T, Benitah SA, Valerón PF, Espina C and Lacal JC: Rho GTPase expression in tumourigenesis: evidence for a significant link. Bioessays. 27:602–613. 2005.PubMed/NCBI
40.	Ellenbroek SI and Collard JG: RhoGTPases: functions and association with cancer. Clin Exp Metastasis. 24:657–672. 2007. View Article : Google Scholar : PubMed/NCBI
41.	Rivero F, Dislich H, Glockner G and Noegel AA: The Dictyostelium discoideum family of Rho-related proteins. Nucleic Acids Res. 29:1068–1079. 2001. View Article : Google Scholar : PubMed/NCBI