Downregulation of miR-518a-3p activates the NIK-dependent NF-κB pathway in colorectal cancer

Qu,L. L.; He,L.; Zhao,X.; Xu,W.

doi:10.3892/ijmm.2015.2145

Downregulation of miR-518a-3p activates the NIK-dependent NF-κB pathway in colorectal cancer

Authors:
- L. L. Qu
- L. He
- X. Zhao
- W. Xu
View Affiliations

Affiliations: Department of Laboratory Medicine, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China, Department of Gastroenterology Surgery, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China, Department of Hepatology, The First Hospital of Jilin University, Changchun, Jilin 130021, P.R. China
Published online on: March 18, 2015 https://doi.org/10.3892/ijmm.2015.2145
Pages: 1266-1272
Copyright: © Qu et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to investigate the biological role and underlying mechanisms of action of miR‑518a‑3p in the progression and invasion of colorectal cancer (CRC). Reverse transcription-quantitative PCR (RT-qPCR) was used to examine the mRNA expression levels of miR‑518a‑3p in 5 CRC cell lines (SW480, SW620, HCT116, HT29 and LoVo) in a normal colonic cell line, NCM460, as well as in tumor tissues with or without metastases. The biological effects of miR‑518a‑3p were assessed in the CRC cell lines by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometric analysis, and RT‑qPCR and western blot analyses were employed to evaluate the expression of miR‑518a‑3p targets. The regulation of NF‑κB‑inducing kinase (NIK) by miR‑518a‑3p was confirmed using luciferase activity assays. Our results revealed that miR‑518a‑3p was significantly downregulated in the CRC cell lines compared with the normal colonic cell line (P<0.05), as well as in the CRC tissues with distant metastases compared with the tissues without metastases. The downregulation of miR‑518a‑3p was associated with tumor size, distant metastasis and TNM stage in the patients with CRC. Moreover, the ectopic expression of miR‑518a‑3p and the inhibition of NIK by RNA interference markedly reduced cell proliferation and enhanced the apoptosis of CRC cells. Further experiments revealed that NIK, a regulator of NF‑κB, was a downstream target of miR‑518a‑3p. The presents findings indicate that miR‑518a‑3p plays an important role in the progression of CRC by targeting NIK.

View Figures

View References

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 : PubMed/NCBI
2	Hironaka N, Mochida K, Mori N, Maeda M, Yamamoto N and Yamaoka S: Tax-independent constitutive IkappaB kinase activation in adult T-cell leukemia cells. Neoplasia. 6:266–278. 2004. View Article : Google Scholar : PubMed/NCBI
3	Prasad S, Ravindran J and Aggarwal BB: NF-kappaB and cancer: How intimate is this relationship. Mol Cell Biochem. 336:25–37. 2010. View Article : Google Scholar
4	Claudius AK, Kankipati CS, Kilari RS, Hassan S, Guest K, Russell ST, Perry CJ, Stark LA and Nicholl ID: Identification of aspirin analogues that repress NF-κB signalling and demonstrate anti-proliferative activity towards colorectal cancer in vitro and in vivo. Oncol Rep. 32:1670–1680. 2014.PubMed/NCBI
5	Yang H, Qi H, Ren J, Cui J, Li Z, Waldum HL and Cui G: Involvement of NF-κB/IL-6 pathway in the processing of colorectal carcinogenesis in colitis mice. Int J Inflam. 2014:1309812014. View Article : Google Scholar
6	Thu YM, Su Y, Yang J, Splittgerber R, Na S, Boyd A, Mosse C, Simons C and Richmond A: NF-κB inducing kinase (NIK) modulates melanoma tumorigenesis by regulating expression of pro-survival factors through the β-catenin pathway. Oncogene. 31:2580–2592. 2012. View Article : Google Scholar :
7	Thu YM and Richmond A: NF-κB inducing kinase: A key regulator in the immune system and in cancer. Cytokine Growth Factor Rev. 21:213–226. 2010. View Article : Google Scholar : PubMed/NCBI
8	Annunziata CM, Davis RE, Demchenko Y, Bellamy W, Gabrea A, Zhan F, Lenz G, Hanamura I, Wright G, Xiao W, et al: Frequent engagement of the classical and alternative NF-kappaB pathways by diverse genetic abnormalities in multiple myeloma. Cancer Cell. 12:115–130. 2007. View Article : Google Scholar : PubMed/NCBI
9	Zhao JL and Starczynowski DT: Role of microRNA-146a in normal and malignant hematopoietic stem cell function. Front Genet. 5:2192014. View Article : Google Scholar : PubMed/NCBI
10	Bandres E, Bitarte N, Arias F, Agorreta J, Fortes P, Agirre X, Zarate R, Diaz-Gonzalez JA, Ramirez N, Sola JJ, et al: microRNA-451 regulates macrophage migration inhibitory factor production and proliferation of gastrointestinal cancer cells. Clin Cancer Res. 15:2281–2290. 2009. View Article : Google Scholar : PubMed/NCBI
11	Chen X, Guo X, Zhang H, Xiang Y, Chen J, Yin Y, Cai X, Wang K, Wang G, Ba Y, et al: Role of miR-143 targeting KRAS in colorectal tumorigenesis. Oncogene. 28:1385–1392. 2009. View Article : Google Scholar : PubMed/NCBI
12	Zhang J, Wu H, Li P, Zhao Y, Liu M and Tang H: NF-κB-modulated miR-130a targets TNF-α in cervical cancer cells. J Transl Med. 12:1552014. View Article : Google Scholar
13	Liu Z, Gersbach E, Zhang X, Xu X, Dong R, Lee P, Liu J, Kong B, Shao C and Wei JJ: miR-106a represses the Rb tumor suppressor p130 to regulate cellular proliferation and differentiation in high-grade serous ovarian carcinoma. Mol Cancer Res. 11:1314–1325. 2013. View Article : Google Scholar : PubMed/NCBI
14	Abdelmohsen K, Srikantan S, Kuwano Y and Gorospe M: miR-519 reduces cell proliferation by lowering RNA-binding protein HuR levels. Proc Natl Acad Sci USA. 105:20297–20302. 2008. View Article : Google Scholar : PubMed/NCBI
15	Farhana L, Dawson MI, Murshed F, Das JK, Rishi AK and Fontana JA: Upregulation of miR-150^* and miR-630 induces apoptosis in pancreatic cancer cells by targeting IGF-1R. PLoS One. 8:e610152013. View Article : Google Scholar
16	Guo J, Li M, Meng X, Sui J, Dou L, Tang W, Huang X, Man Y, Wang S and Li J: MiR-291b-3p induces apoptosis in liver cell line NCTC1469 by reducing the level of RNA-binding protein HuR. Cell Physiol Biochem. 33:810–822. 2014. View Article : Google Scholar : PubMed/NCBI
17	Odqvist L, Sánchez-Beato M, Montes-Moreno S, et al: NIK controls classical and alternative NF-κB activation and is necessary for the survival of human T-cell lymphoma cells. Clin Cancer Res. 19:2319–2330. 2013. View Article : Google Scholar : PubMed/NCBI
18	Li M, Wang X, Liu M, Qi X and Li J: NF-κB signaling inhibition and anticancer activities of LLDT-246 on human colorectal cancer HCT-116 cells in vitro. Biomed Pharmacother. 68:527–535. 2014. View Article : Google Scholar : PubMed/NCBI
19	Hajder J, Marisavljević D, Stanisavljević N, Mihaljević B, Kovcin V, Marković O and Zivković R: BCL10 aberations and NF-kappa B activation involving p65 are absent or rare in primary gastric MALT lymphoma. Vojnosanit Pregl. 71:1040–1044. 2014. View Article : Google Scholar : PubMed/NCBI
20	Celegato M, Borghese C, Casagrande N, Carbone A, Colombatti A and Aldinucci D: Bortezomib down-modulates the survival factor interferon regulatory factor 4 in Hodgkin lymphoma cell lines and decreases the protective activity of Hodgkin lymphoma-associated fibroblasts. Leuk Lymphoma. 55:149–159. 2014. View Article : Google Scholar
21	Hayden MS and Ghosh S: Shared principles in NF-kappaB signaling. Cell. 132:344–362. 2008. View Article : Google Scholar : PubMed/NCBI
22	Kupcinskas J, Bruzaite I, Juzenas S, Gyvyte U, Jonaitis L, Kiudelis G, Skieceviciene J, Leja M, Pauzas H, Tamelis A, et al: Lack of association between miR-27a, miR-146a, miR-196a-2, miR-492 and miR-608 gene polymorphisms and colorectal cancer. Sci Rep. 4:59932014. View Article : Google Scholar : PubMed/NCBI
23	Rodríguez-Montes JA and Menéndez Sánchez P: Role of micro-RNA in colorectal cancer screening. Cir Esp. 92:654–658. 2014. View Article : Google Scholar : PubMed/NCBI
24	Valeri N, Gasparini P, Braconi C, Paone A, Lovat F, Fabbri M, Sumani KM, Alder H, Amadori D, Patel T, et al: MicroRNA-21 induces resistance to 5-fluorouracil by down-regulating human DNA MutS homolog 2 (hMSH2). Proc Natl Acad Sci USA. 107:21098–21103. 2010. View Article : Google Scholar : PubMed/NCBI
25	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
26	Boni V, Bitarte N, Cristobal I, Zarate R, Rodriguez J, Maiello E, Garcia-Foncillas J and Bandres E: miR-192/miR-215 influence 5-fluorouracil resistance through cell cycle-mediated mechanisms complementary to its post-transcriptional thymidilate synthase regulation. Mol Cancer Ther. 9:2265–2275. 2010. View Article : Google Scholar : PubMed/NCBI
27	Wang Z, Sun X, Wang Y, Liu X, Xuan Y and Hu S: Association between miR-27a genetic variants and susceptibility to colorectal cancer. Diagn Pathol. 9:1462014. View Article : Google Scholar : PubMed/NCBI
28	Wang S, Yang MH, Wang XY, Lin J and Ding YQ: Increased expression of miRNA-182 in colorectal carcinoma: An independent and tissue-specific prognostic factor. Int J Clin Exp Pathol. 7:3498–3503. 2014.PubMed/NCBI
29	Huh JH, Kim TH, Kim K, Song JA, Jung YJ, Jeong JY, Lee MJ, Kim YK, Lee DH and An HJ: Dysregulation of miR-106a and miR-591 confers paclitaxel resistance to ovarian cancer. Br J Cancer. 109:452–461. 2013. View Article : Google Scholar : PubMed/NCBI
30	Plummer SM, Holloway KA, Manson MM, Munks RJ, Kaptein A, Farrow S and Howells L: Inhibition of cyclo-oxygenase 2 expression in colon cells by the chemopreventive agent curcumin involves inhibition of NF-kappaB activation via the NIK/IKK signalling complex. Oncogene. 18:6013–6020. 1999. View Article : Google Scholar : PubMed/NCBI