Top

Journal of Experimental & Clinical Cancer Research

Published in:

Open Access 01-12-2014 | Research

MicroRNA-320c inhibits tumorous behaviors of bladder cancer by targeting Cyclin-dependent kinase 6

Authors: Xiao Wang, Jian Wu, Yiwei Lin, Yi Zhu, Xianglai Xu, Xin Xu, Zhen Liang, Shiqi Li, Zhenghui Hu, Xiangyi Zheng, Liping Xie

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2014

Abstract

Background

Increasing evidence has suggested that dysregulation of microRNAs (miRNAs) could contribute to human disease including cancer. Previous miRNA microarray analysis illustrated that miR-320c is down-regulated in various cancers. However, the roles of miR-320c in human bladder cancer have not been well elucidated. Therefore, this study was performed to investigate the biological functions and molecular mechanisms of miR-320c in human bladder cancer cell lines, discussing whether it could be a therapeutic biomarker of bladder cancer in the future.

Methods

Two human bladder cancer cell lines and samples from thirteen patients with bladder cancer were analyzed for the expression of miR-320c by quantitative RT-PCR. Over-expression of miR-320c was established by transfecting mimics into T24 and UM-UC-3. Cell proliferation and cell cycle were assessed by cell viability assay, flow cytometry and colony formation assay. Cell motility ability was evaluated by transwell assay. The target gene of miR-320c was determined by luciferase assay, quantitative RT-PCR and western blot. The regulation of cell cycle and mobility by miR-320c was analyzed by western blot.

Results

We observed that miR-320c was down-regulated in human bladder cancer tissues and bladder cancer cell lines T24 and UM-UC-3. Over-expression of miR-320c could induce G1 phase arrest in UM-UC-3 and T24 cells, and subsequently inhibited cell growth. We also indentified miR-320c could impair UM-UC-3 and T24 cell motility. In addition, we identified CDK6, a cell cycle regulator, as a novel target of miR-320c. Moreover, we demonstrated miR-320c could induce bladder cancer cell cycle arrest and mobility via regulating CDK6. We also observed that inhibition of miR-320c or restoration of CDK6 in miR-320c-over-expressed bladder cancer cells partly reversed the suppressive effects of miR-320c.

Conclusions

miR-320c could inhibit the proliferation, migration and invasion of bladder cancer cells via regulating CDK6. Our study revealed that miR-320c could be a therapeutic biomarker of bladder cancer in the future.

Available only for authorised users

Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin. 2011, 61 (2): 69-90. 10.3322/caac.20107.CrossRefPubMed

Siegel R, Ma J, Zou Z, Jemal A: Cancer statistics, 2014. CA Cancer J Clin. 2014, 64 (1): 9-29. 10.3322/caac.21208.CrossRefPubMed

Wang X, Lin YW, Wang S, Wu J, Mao QQ, Zheng XY, Xie LP: A meta-analysis of tea consumption and the risk of bladder cancer. Urol Int. 2013, 90 (1): 10-16. 10.1159/000342804.CrossRefPubMed

Chiong E, Kesavan A, Mahendran R, Chan YH, Sng JH, Lim YK, Kamaraj R, Tan TM, Esuvaranathan K: NRAMP1 and hGPX1 gene polymorphism and response to bacillus Calmette-Guerin therapy for bladder cancer. Eur Urol. 2011, 59 (3): 430-437. 10.1016/j.eururo.2010.11.031.CrossRefPubMed

Casadio V, Molinari C, Calistri D, Tebaldi M, Gunelli R, Serra L, Falcini F, Zingaretti C, Silvestrini R, Amadori D, Zoli W: DNA Methylation profiles as predictors of recurrence in non muscle invasive bladder cancer: an MS-MLPA approach. J Exp Clin Cancer Res. 2013, 32: 94-10.1186/1756-9966-32-94.PubMedCentralCrossRefPubMed

Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004, 116 (2): 281-297. 10.1016/S0092-8674(04)00045-5.CrossRefPubMed

Bartel DP: MicroRNAs: target recognition and regulatory functions. Cell. 2009, 136 (2): 215-233. 10.1016/j.cell.2009.01.002.PubMedCentralCrossRefPubMed

Calin GA, Liu CG, Sevignani C, Ferracin M, Felli N, Dumitru CD, Shimizu M, Cimmino A, Zupo S, Dono M, Dell'Aquila ML, Alder H, Rassenti L, Kipps TJ, Bullrich F, Negrini M, Croce CM: MicroRNA profiling reveals distinct signatures in B cell chronic lymphocytic leukemias. Proc Natl Acad Sci U S A. 2004, 101 (32): 11755-11760. 10.1073/pnas.0404432101.PubMedCentralCrossRefPubMed

Zhou B, Chen H, Wei D, Kuang Y, Zhao X, Li G, Xie J, Chen P: A novel miR-219-SMC4-JAK2/Stat3 regulatory pathway in human hepatocellular carcinoma. J Exp Clin Cancer Res. 2014, 33 (1): 55-10.1186/1756-9966-33-55.PubMedCentralCrossRefPubMed

10.

Ma C, Nong K, Wu B, Dong B, Bai Y, Zhu H, Wang W, Huang X, Yuan Z, Ai K: miR-212 promotes pancreatic cancer cell growth and invasion by targeting the hedgehog signaling pathway receptor patched-1. J Exp Clin Cancer Res. 2014, 33: 54-10.1186/1756-9966-33-54.PubMedCentralCrossRefPubMed

11.

Wang Z, Wang J, Yang Y, Hao B, Wang R, Li Y, Wu Q: Loss of has-miR-337-3p expression is associated with lymph node metastasis of human gastric cancer. J Exp Clin Cancer Res. 2013, 32: 76-10.1186/1756-9966-32-76.PubMedCentralCrossRefPubMed

12.

Li Y, Chao Y, Fang Y, Wang J, Wang M, Zhang H, Ying M, Zhu X, Wang H: MTA1 promotes the invasion and migration of non-small cell lung cancer cells by downregulating miR-125b. J Exp Clin Cancer Res. 2013, 32: 33-10.1186/1756-9966-32-33.PubMedCentralCrossRefPubMed

13.

Gao SM, Yang JJ, Chen CQ, Chen JJ, Ye LP, Wang LY, Wu JB, Xing CY, Yu K: Pure curcumin decreases the expression of WT1 by upregulation of miR-15a and miR-16-1 in leukemic cells. J Exp Clin Cancer Res. 2012, 31: 27-10.1186/1756-9966-31-27.PubMedCentralCrossRefPubMed

14.

Catto JW, Alcaraz A, Bjartell AS, De Vere WR, Evans CP, Fussel S, Hamdy FC, Kallioniemi O, Mengual L, Schlomm T, Visakorpi T: MicroRNA in prostate, bladder, and kidney cancer: a systematic review. Eur Urol. 2011, 59 (5): 671-681. 10.1016/j.eururo.2011.01.044.CrossRefPubMed

15.

Gottardo F, Liu CG, Ferracin M, Calin GA, Fassan M, Bassi P, Sevignani C, Byrne D, Negrini M, Pagano F, Gomella LG, Croce CM, Baffa R: Micro-RNA profiling in kidney and bladder cancers. Urol Oncol. 2007, 25 (5): 387-392. 10.1016/j.urolonc.2007.01.019.CrossRefPubMed

16.

Yan LX, Huang XF, Shao Q, Huang MY, Deng L, Wu QL, Zeng YX, Shao JY: MicroRNA miR-21 overexpression in human breast cancer is associated with advanced clinical stage, lymph node metastasis and patient poor prognosis. RNA. 2008, 14 (11): 2348-2360. 10.1261/rna.1034808.PubMedCentralCrossRefPubMed

17.

Schepeler T, Reinert JT, Ostenfeld MS, Christensen LL, Silahtaroglu AN, Dyrskjot L, Wiuf C, Sorensen FJ, Kruhoffer M, Laurberg S, Kauppinen S, Orntoft TF, Andersen CL: Diagnostic and prognostic microRNAs in stage II colon cancer. Cancer Res. 2008, 68 (15): 6416-6424. 10.1158/0008-5472.CAN-07-6110.CrossRefPubMed

18.

Schaar DG, Medina DJ, Moore DF, Strair RK, Ting Y: miR-320 targets transferrin receptor 1 (CD71) and inhibits cell proliferation. Exp Hematol. 2009, 37 (2): 245-255. 10.1016/j.exphem.2008.10.002.CrossRefPubMed

19.

Hsieh IS, Chang KC, Tsai YT, Ke JY, Lu PJ, Lee KH, Yeh SD, Hong TM, Chen YL: MicroRNA-320 suppresses the stem cell-like characteristics of prostate cancer cells by downregulating the Wnt/beta-catenin signaling pathway. Carcinogenesis. 2013, 34 (3): 530-538. 10.1093/carcin/bgs371.CrossRefPubMed

20.

Yao J, Liang LH, Zhang Y, Ding J, Tian Q, Li JJ, He XH: GNAI1 Suppresses Tumor Cell Migration and Invasion and is Post-Transcriptionally Regulated by Mir-320a/c/d in Hepatocellular Carcinoma. Cancer Biol Med. 2012, 9 (4): 234-241.PubMedCentralPubMed

21.

Iwagami Y, Eguchi H, Nagano H, Akita H, Hama N, Wada H, Kawamoto K, Kobayashi S, Tomokuni A, Tomimaru Y, Mori M, Doki Y: miR-320c regulates gemcitabine-resistance in pancreatic cancer via SMARCC1. Br J Cancer. 2013, 109 (2): 502-511. 10.1038/bjc.2013.320.PubMedCentralCrossRefPubMed

22.

Zhu Y, Lu Y, Zhang Q, Liu JJ, Li TJ, Yang JR, Zeng C, Zhuang SM: MicroRNA-26a/b and their host genes cooperate to inhibit the G1/S transition by activating the pRb protein. Nucleic Acids Res. 2012, 40 (10): 4615-4625. 10.1093/nar/gkr1278.PubMedCentralCrossRefPubMed

23.

Chen X, Wang X, Ruan A, Han W, Zhao Y, Lu X, Xiao P, Shi H, Wang R, Chen L, Chen S, Du Q, Yang H, Zhang X: miR-141 is a key regulator of renal cell carcinoma proliferation and metastasis by controlling EphA2 expression. Clin Cancer Res. 2014, 20 (10): 2617-2630. 10.1158/1078-0432.CCR-13-3224.CrossRefPubMed

24.

Zhu X, Li Y, Shen H, Li H, Long L, Hui L, Xu W: miR-137 inhibits the proliferation of lung cancer cells by targeting Cdc42 and Cdk6. FEBS Lett. 2013, 587 (1): 73-81. 10.1016/j.febslet.2012.11.004.CrossRefPubMed

25.

Lapointe J, Lachance Y, Labrie Y, Labrie C: A p18 mutant defective in CDK6 binding in human breast cancer cells. Cancer Res. 1996, 56 (20): 4586-4589.PubMed

26.

Wang G, Zheng L, Yu Z, Liao G, Lu L, Xu R, Zhao Z, Chen G: Increased cyclin-dependent kinase 6 expression in bladder cancer. Oncol Lett. 2012, 4 (1): 43-46.PubMedCentralPubMed

27.

Prasad SM, Decastro GJ, Steinberg GD: Urothelial carcinoma of the bladder: definition, treatment and future efforts. Nat Rev Urol. 2011, 8 (11): 631-642. 10.1038/nrurol.2011.144.CrossRefPubMed

28.

Koturbash I, Zemp FJ, Pogribny I, Kovalchuk O: Small molecules with big effects: the role of the microRNAome in cancer and carcinogenesis. Mutat Res. 2011, 722 (2): 94-105. 10.1016/j.mrgentox.2010.05.006.CrossRefPubMed

29.

Zhang M, Yang Q, Zhang L, Zhou S, Ye W, Yao Q, Li Z, Huang C, Wen Q, Wang J: miR-302b is a potential molecular marker of esophageal squamous cell carcinoma and functions as a tumor suppressor by targeting ErbB4. J Exp Clin Cancer Res. 2014, 33: 10-10.1186/1756-9966-33-10.PubMedCentralCrossRefPubMed

30.

Yang N, Kaur S, Volinia S, Greshock J, Lassus H, Hasegawa K, Liang S, Leminen A, Deng S, Smith L, Johnstone CN, Chen XM, Liu CG, Huang Q, Katsaros D, Calin GA, Weber BL, Butzow R, Croce CM, Coukos G, Zhang L: MicroRNA microarray identifies Let-7i as a novel biomarker and therapeutic target in human epithelial ovarian cancer. Cancer Res. 2008, 68 (24): 10307-10314. 10.1158/0008-5472.CAN-08-1954.PubMedCentralCrossRefPubMed

31.

Lin Y, Chen H, Hu Z, Mao Y, Xu X, Zhu Y, Wu J, Li S, Mao Q, Zheng X, Xie L: miR-26a inhibits proliferation and motility in bladder cancer by targeting HMGA1. FEBS Lett. 2013, 587 (15): 2467-2473. 10.1016/j.febslet.2013.06.021.CrossRefPubMed

32.

Li S, Xu X, Hu Z, Wu J, Zhu Y, Chen H, Mao Y, Lin Y, Luo J, Zheng X, Xie L: MicroRNA-490-5p inhibits proliferation of bladder cancer by targeting c-Fos. Biochem Biophys Res Commun. 2013, 441 (4): 976-981. 10.1016/j.bbrc.2013.11.006.CrossRefPubMed

33.

Landis MW, Pawlyk BS, Li T, Sicinski P, Hinds PW: Cyclin D1-dependent kinase activity in murine development and mammary tumorigenesis. Cancer Cell. 2006, 9 (1): 13-22. 10.1016/j.ccr.2005.12.019.CrossRefPubMed

34.

Zhang Z, Huang L, Yu Z, Chen X, Yang D, Zhan P, Dai M, Huang S, Han Z, Cao K: Let-7a functions as a tumor suppressor in Ewing's sarcoma cell lines partly by targeting cyclin-dependent kinase 6. DNA Cell Biol. 2014, 33 (3): 136-147. 10.1089/dna.2013.2179.PubMedCentralCrossRefPubMed

35.

Lamb R, Lehn S, Rogerson L, Clarke RB, Landberg G: Cell cycle regulators cyclin D1 and CDK4/6 have estrogen receptor-dependent divergent functions in breast cancer migration and stem cell-like activity. Cell Cycle. 2013, 12 (15): 2384-2394. 10.4161/cc.25403.PubMedCentralCrossRefPubMed

36.

Wang C, Lisanti MP, Liao DJ: Reviewing once more the c-myc and Ras collaboration: converging at the cyclin D1-CDK4 complex and challenging basic concepts of cancer biology. Cell Cycle. 2011, 10 (1): 57-67. 10.4161/cc.10.1.14449.PubMedCentralCrossRefPubMed

Title: MicroRNA-320c inhibits tumorous behaviors of bladder cancer by targeting Cyclin-dependent kinase 6
Authors: Xiao Wang
Jian Wu
Yiwei Lin
Yi Zhu
Xianglai Xu
Xin Xu
Zhen Liang
Shiqi Li
Zhenghui Hu
Xiangyi Zheng
Liping Xie
Publication date: 01-12-2014
Publisher: BioMed Central
Published in: Journal of Experimental & Clinical Cancer Research / Issue 1/2014
Electronic ISSN: 1756-9966
DOI: https://doi.org/10.1186/s13046-014-0069-6

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2014

The role of microRNA-133b and its target gene FSCN1 in gastric cancer

Proteins of the VEGFR and EGFR pathway as predictive markers for adjuvant treatment in patients with stage II/III colorectal cancer: results of the FOGT-4 trial

Quantitative assessment of the influence of EPHX1 gene polymorphisms and cancer risk: a meta-analysis with 94,213 subjects

Reciprocal regulation of PCGEM1 and miR-145promote proliferation of LNCaP prostate cancer cells

Store-operated Ca2+ entry regulates glioma cell migration and invasion via modulation of Pyk2 phosphorylation

Design and validation of an oligonucleotide microarray for the detection of genomic rearrangements associated with common hereditary cancer syndromes

Keynote webinar | Spotlight on antibody–drug conjugates in cancer