Klf4 inhibits tumor growth and metastasis by targeting microRNA-31 in human hepatocellular carcinoma

Tian,Chuan; Yao,Shanshan; Liu,Li; Ding,Youcheng; Ye,Qingwang; Dong,Xiao; Gao,Yong; Yang,Ning; Li,Qi

doi:10.3892/ijmm.2016.2812

Klf4 inhibits tumor growth and metastasis by targeting microRNA-31 in human hepatocellular carcinoma

Authors:
- Chuan Tian
- Shanshan Yao
- Li Liu
- Youcheng Ding
- Qingwang Ye
- Xiao Dong
- Yong Gao
- Ning Yang
- Qi Li
View Affiliations

Affiliations: Department of Oncology, Shanghai First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai, P.R. China, Department of Pharmacy, Guiyang Hospital of Guizhou Aviation Industry Group, Guiyang, Guizhou, P.R. China, Department of General Surgery, Shanghai East Hospital, Tongji University, Shanghai, P.R. China, Department of Liver Surgery, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, P.R. China, Department of Oncology, Shanghai East Hospital, Tongji University, Shanghai, P.R. China
Published online on: November 24, 2016 https://doi.org/10.3892/ijmm.2016.2812
Pages: 47-56
Copyright: © Tian et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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

MicroRNAs (miRNAs or miRs) are short, endogenous non-coding RNA molecules, demonstrating abnormal expression in cancer initiation and progression. In this study, we profiled 18 differentially regulated miRNAs, including miRNA‑31, using miRNA array. Kruppel (or Krüppel)-like factor 4 (Klf4) is a transcription factor and putative tumor suppressor. Both were found to be significantly downregulated in liver cancer tissues and cells. However, little is known about the correlation between Klf4 and miRNA‑31 in hepatocellular carcinoma (HCC). The mRNA expression of Klf4 was decreased and inversely associated with the clinical stage, T classification and hepatitis B in patients with HCC, while the expression of miR‑31 was lower (r=0.326, P=0.018). Using cell counting kit 8 (CCK8) and Transwell migration assays, we found that Klf4 and miR‑31 inhibited the proliferation and metastasis of liver cancer cells. Moreover, we demonstrated that Klf4 directly binds to the promoter of miR‑31 and activates its transcription. In vitro experiments confirmed that Klf4 regulated miR‑31 and thereby inhibited HCC cell growth and metastasis. Taken together, our findings indicate that Klf4 directly regulates miR‑31 in HCC. Thus, miR-31 may serve as a potential diagnostic marker and therapeutic target in HCC.

View Figures

View References

1	Maluccio M and Covey A: Recent progress in understanding, diagnosing, and treating hepatocellular carcinoma. CA Cancer J Clin. 62:394–399. 2012. View Article : Google Scholar : PubMed/NCBI
2	Bhayani NH, Jiang Y, Hamed O, Kimchi ET, Staveley-O'Carroll KF and Gusani NJ: Advances in the pharmacologic treatment of hepatocellular carcinoma. Curr Clin Pharmacol. 10:299–304. 2015. View Article : Google Scholar : PubMed/NCBI
3	Moeini A, Cornellà H and Villanueva A: Emerging signaling pathways in hepatocellular carcinoma. Liver Cancer. 1:83–93. 2012. View Article : Google Scholar
4	Lujambio A and Lowe SW: The microcosmos of cancer. Nature. 482:347–355. 2012. View Article : Google Scholar : PubMed/NCBI
5	Di Leva G, Garofalo M and Croce CM: MicroRNAs in cancer. Annu Rev Pathol. 9:287–314. 2014. View Article : Google Scholar :
6	Fang W and Bartel DP: The menu of features that define primary microRNAs and enable de novo design of dicroRNA genes. Mol Cell. 60:131–145. 2015. View Article : Google Scholar : PubMed/NCBI
7	Parpart S, Roessler S, Dong F, Rao V, Takai A, Ji J, Qin LX, Ye QH, Jia HL, Tang ZY and Wang XW: Modulation of miR-29 expression by α-fetoprotein is linked to the hepatocellular carcinoma epigenome. Hepatology. 60:872–883. 2014. View Article : Google Scholar : PubMed/NCBI
8	Najafi Z, Sharifi M and Javadi G: Degradation of miR-21 induces apoptosis and inhibits cell proliferation in human hepatocellular carcinoma. Cancer Gene Ther. 22:530–535. 2015. View Article : Google Scholar : PubMed/NCBI
9	He XX, Guo AY, Xu CR, Chang Y, Xiang GY, Gong J, Dan ZL, Tian DA, Liao JZ and Lin JS: Bioinformatics analysis identifies miR-221 as a core regulator in hepatocellular carcinoma and its silencing suppresses tumor properties. Oncol Rep. 32:1200–1210. 2014.PubMed/NCBI
10	Yang X, Liang L, Zhang XF, Jia HL, Qin Y, Zhu XC, Gao XM, Qiao P, Zheng Y, Sheng YY, et al: MicroRNA-26a suppresses tumor growth and metastasis of human hepatocellular carcinoma by targeting interleukin-6-Stat3 pathway. Hepatology. 58:158–170. 2013. View Article : Google Scholar : PubMed/NCBI
11	Baek S, Cho KJ, Ju HL, Moon H, Choi SH, Chung SI, Park JY, Choi KH, Kim Y, Ahn SH, et al: Analysis of miRNA expression patterns in human and mouse hepatocellular carcinoma cells. Hepatol Res. 45:1331–1340. 2015. View Article : Google Scholar : PubMed/NCBI
12	Mlcochova J, Faltejskova-Vychytilova P, Ferracin M, Zagatti B, Radova L, Svoboda M, Nemecek R, John S, Kiss I, Vyzula R, et al: MicroRNA expression profiling identifies miR-31-5p/3p as associated with time to progression in wild-type RAS metastatic colorectal cancer treated with cetuximab. Oncotarget. 6:38695–38704. 2015.PubMed/NCBI
13	Tseng SH, Yang CC, Yu EH, Chang C, Lee YS, Liu CJ, Chang KW and Lin SC: K14-EGFP-miR-31 transgenic mice have high susceptibility to chemical-induced squamous cell tumorigenesis that is associating with Ku80 repression. Int J Cancer. 136:1263–1275. 2015. View Article : Google Scholar
14	Lu WC, Liu CJ, Tu HF, Chung YT, Yang CC, Kao SY, Chang KW and Lin SC: miR-31 targets ARID1A and enhances the oncogenicity and stemness of head and neck squamous cell carcinoma. Oncotarget. Aug 9–2016.Epub ahead of print.
15	Gao W, Liu L, Xu J, Shao Q, Liu Y, Zeng H and Shu Y: A systematic analysis of predicted MiR-31-targets identifies a diagnostic and prognostic signature for lung cancer. Biomed Pharmacother. 68:419–427. 2014. View Article : Google Scholar : PubMed/NCBI
16	Taccioli C, Garofalo M, Chen H, Jiang Y, Tagliazucchi GM, Di Leva G, Alder H, Fadda P, Middleton J, Smalley KJ, et al: Repression of esophageal neoplasia and inflammatory signaling by anti-miR-31 delivery in vivo. J Natl Cancer Inst. 107:djv2202015. View Article : Google Scholar : PubMed/NCBI
17	Wang S, Li Q, Wang K, Dai Y, Yang J, Xue S, Han F, Zhang Q, Liu J and Wu W: Decreased expression of microRNA-31 associates with aggressive tumor progression and poor prognosis in patients with bladder cancer. Clin Transl Oncol. 15:849–854. 2013. View Article : Google Scholar : PubMed/NCBI
18	Schaefer A, Jung M, Mollenkopf HJ, Wagner I, Stephan C, Jentzmik F, Miller K, Lein M, Kristiansen G and Jung K: Diagnostic and prognostic implications of microRNA profiling in prostate carcinoma. Int J Cancer. 126:1166–1176. 2010.
19	Ruoming W, Zhen Y, Tengteng Z and Jisheng H: Tumor suppressor microRNA-31 inhibits gastric carcinogenesis by targeting Smad4 and SGPP2. Cancer Gene Ther. 22:564–572. 2015. View Article : Google Scholar : PubMed/NCBI
20	Kim HS, Lee KS, Bae HJ, Eun JW, Shen Q, Park SJ, Shin WC, Yang HD, Park M, Park WS, et al: MicroRNA-31 functions as a tumor suppressor by regulating cell cycle and epithelial-mesenchymal transition regulatory proteins in liver cancer. Oncotarget. 6:8089–8102. 2015. View Article : Google Scholar : PubMed/NCBI
21	Viré E, Curtis C, Davalos V, Git A, Robson S, Villanueva A, Vidal A, Barbieri I, Aparicio S, Esteller M, et al: The breast cancer oncogene EMSY represses transcription of antimetastatic microRNA miR-31. Mol Cell. 53:806–818. 2014. View Article : Google Scholar : PubMed/NCBI
22	Creighton CJ, Fountain MD, Yu Z, Nagaraja AK, Zhu H, Khan M, Olokpa E, Zariff A, Gunaratne PH, Matzuk MM and Anderson ML: Molecular profiling uncovers a p53-associated role for microRNA-31 in inhibiting the proliferation of serous ovarian carcinomas and other cancers. Cancer Res. 70:1906–1915. 2010. View Article : Google Scholar : PubMed/NCBI
23	Zhang N, Zhang J, Shuai L, Zha L, He M, Huang Z and Wang Z: Krüppel-like factor 4 negatively regulates β-catenin expression and inhibits the proliferation, invasion and metastasis of gastric cancer. Int J Oncol. 40:2038–2048. 2012.PubMed/NCBI
24	Li Q, Gao Y, Jia Z, Mishra L, Guo K, Li Z, Le X, Wei D, Huang S and Xie K: Dysregulated Krüppel-like factor 4 and vitamin D receptor signaling contribute to progression of hepatocellular carcinoma. Gastroenterology. 143:799–810.e1. 22012. View Article : Google Scholar
25	Yu T, Chen X, Lin T, Liu J, Li M, Zhang W, Xu X, Zhao W, Liu M, Napier DL, et al: KLF4 deletion alters gastric cell lineage and induces MUC2 expression. Cell Death Dis. 7:e22552016. View Article : Google Scholar : PubMed/NCBI
26	Lv H, Zhang Z, Wang Y, Li C, Gong W and Wang X: MicroRNA-92a promotes colorectal cancer cell growth and migration by inhibiting KLF4. Oncol Res. 23:283–290. 2016. View Article : Google Scholar : PubMed/NCBI
27	Liu S, Yang H, Chen Y, He B and Chen Q: Krüppel-like factor 4 enhances sensitivity of cisplatin to lung cancer cells and inhibits regulating epithelial-to-mMesenchymal transition. Oncol Res. 24:81–87. 2016. View Article : Google Scholar
28	Gandellini P, Giovannetti E and Nicassio F: MicroRNAs in cancer management: Big challenges for small molecules. Biomed Res Int. 2015:9821562015. View Article : Google Scholar : PubMed/NCBI
29	Zhou J, Lai PB and Tsui SK: Identification of a non-coding KLF4 transcript generated from intron retention and downregulated in human hepatocellular carcinoma. Int J Oncol. 47:1554–1562. 2015.PubMed/NCBI
30	Tien YT, Chang MH, Chu PY, Lin CS, Liu CH and Liao AT: Downregulation of the KLF4 transcription factor inhibits the proliferation and migration of canine mammary tumor cells. Vet J. 205:244–253. 2015. View Article : Google Scholar : PubMed/NCBI
31	Mao Q, Quan T, Luo B, Guo X, Liu L and Zheng Q: MiR-375 targets Klf4 and impacts the proliferation of colorectal carcinoma. Tumour Biol. 37:463–471. 2016. View Article : Google Scholar
32	Lin ZS, Chu HC, Yen YC, Lewis BC and Chen YW: Krüppel-like factor 4, a tumor suppressor in hepatocellular carcinoma cells reverts epithelial mesenchymal transition by suppressing slug expression. PLoS One. 7. pp. e435932012, View Article : Google Scholar
33	Fiorenza A, Lopez-Atalaya JP, Rovira V, Scandaglia M, Geijo-Barrientos E and Barco A: Blocking miRNA biogenesis in adult forebrain neurons enhances seizure susceptibility, fear memory, and food intake by increasing neuronal responsiveness. Cereb Cortex. 26:1619–1633. 2016. View Article : Google Scholar
34	Meehan K and Vella LJ: The contribution of tumour-derived exosomes to the hallmarks of cancer. Crit Rev Clin Lab Sci. 53:121–131. 2016. View Article : Google Scholar
35	Yang F, Yin Y, Wang F, Wang Y, Zhang L, Tang Y and Sun S: miR-17-5p Promotes migration of human hepatocellular carcinoma cells through the p38 mitogen-activated protein kinase-heat shock protein 27 pathway. Hepatology. 51:1614–1623. 2010. View Article : Google Scholar : PubMed/NCBI