a Department of Urology, Zhoupu Hospital, Medicine and Health Sciences, Shanghai University, Shanghai, China b Department of Urology, Shanghai East Hospital, School of Medicine, Tongji University, Shanghai, China
Objective: Deleted in liver cancer 1 (DLC1) is a GTPase-activating protein that is reported as a suppressor in certain human cancers. However, the detailed biological function of DLC1 is still unclear in human prostate cancer (PCa). In the present study, we aimed to explore the function of DLC1 in PCa cells.
Methods: Silencing and overexpression of DLC1 were induced in an androgen-sensitive PCa cell line (LNCaP) using RNA interference and lentiviral vector transduction. The Cell Counting Kit-8 assay was performed to determine cell proliferation. The cell cycle was examined by performing a propidium iodide staining assay.
Results:Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of LNCaP cells. Moreover, DLC1 expression was negatively correlated with Rho-associated protein kinase (ROCK) expression in LNCaP cells. Importantly, this study showed that the ROCK inhibitor Y27632 restored the function of DLC1 in LNCaP cells and reduced the tumorigenicity of LNCaP cells in vivo.
Conclusion: Our results indicated that DLC1 overexpression markedly suppressed the proliferation and cell cycle progression of PCa cells and negatively correlated with ROCK expression in PCa cells and tissue.
. [J]. Asian Journal of Urology, 2023, 10(1): 50-57.
Hua Gong,Kang Chen,Lan Zhou,Yongchao Jin,Weihua Chen. Deleted in liver cancer 1 suppresses the growth of prostate cancer cells through inhibiting Rho-associated protein kinase pathway. Asian Journal of Urology, 2023, 10(1): 50-57.
Siemens DR, Hoare D, Skinner T, Black A. Serum follicle-stimulating hormone levels predict time to development of castration resistant prostate cancer. Can Urol Assoc J 2015; 9: 122e7.
[2]
Sebesta EM, Anderson CB. The surgical management of pros-tate cancer. Semin Oncol 2018; 44:347e57.
doi: 10.1053/j.seminoncol.2018.01.003
[3]
Wong CC, Wong CM, Ko FC, Chan LK, Ching YP, Yam JW, et al. Deleted in liver cancer 1 (DLC1) negatively regulates Rho/ROCK/MLC pathway in hepatocellular carcinoma. PLoS One 2008;3:e2779. https://doi.org/10.1371/journal.pone.0002779.
[4]
Basak P, Dillon R, Leslie H, Raouf A, Mowat MR. The deleted in liver cancer 1 (Dlc1) tumor suppressor is haploinsuf?cient for mammary gland development and epithelial cell polarity. BMC Cancer 2015;15:630. https://doi.org/10.1186/s12885-015-1642-x.
[5]
Su Y, Lin L, Zhang J, Jiang Y, Pan C, Sun L, et al. Low expression of DLC1 is predictive of poor therapeutic ef?ciency of fluoropyrimidine and oxaliplatin as adjuvant chemotherapy in gastric cancer. Mol Med Rep 2015; 12:5771e9.
doi: 10.3892/mmr.2015.4173
[6]
Tripathi V, Popescu NC, Zimonjic DB. DLC DLC1 induces expression of E-cadherin in prostate cancer cells through Rho pathway and suppresses invasion. Oncogene 2014; 33:724e33.
[7]
Hall A. Signal transduction through small GTPasesAdtale of two GAPs. Cell 1992; 69:369e91.
[8]
Amin E, Dubey BN, Zhang SC, Gremer L, Dvorsky R, Moll JM, et al. Rho-kinase: regulation, (dys)function, and inhibition. Biol Chem 2013; 394:1399e410.
doi: 10.1515/hsz-2013-0181
pmid: 23950574
[9]
Matsubara M, Bissell MJ. Inhibitors of rho kinase (ROCK) signaling revert the malignant phenotype of breast cancer cells in 3D context. Oncotarget 2016; 7:31602e22.
doi: 10.18632/oncotarget.9395
pmid: 27203208
[10]
Mu D, Zhou G, Li J, Su B, Guo H. Ursolic acid activates the apoptosis of prostate cancer via ROCK/PTEN mediated mitochondrial translocation of co?lin-1. Oncol Lett 2018; 15: 3202e6.
[11]
Wong CM, Lee JMF, Ching YP, Jin DY, Ng IOL. Genetic and epigenetic alterations of DLC-1 gene in hepatocellular Carci-noma. Cancer Res 2003; 63:7646e51.
[12]
Yang J, Mowry LE, Nejak-Bowen KN, Okabe H, Diegel CR, Lang RA, et al. b-catenin signaling in murine liver zonation and regener-ation: a Wnt-Wnt situation. Hepatology 2014; 60:964e76.
doi: 10.1002/hep.27082
[13]
Cui C, Zhou X, Zhang W, Qu Y, Ke X. Is b-catenin a druggable target for cancer therapy? Trends Biochem Sci 2018; 43: 623e34.
doi: 10.1016/j.tibs.2018.06.003
[14]
Xu J, Prosperi JR, Choudhury N, Olopade OI, Goss KH. Beta-catenin is required for the tumorigenic behavior of triple-negative breast cancer cells. PLoS One 2015; 10:e0117097. https://doi.org/10.1371/journal.pone.0117097.
doi: 10.1371/journal.pone.0117097
[15]
Dong L, Deng J, Sun ZM, Pan AP, Xiang XJ, Zhang L, et al. Interference with the beta-catenin gene in gastric cancer in-duces changes to the miRNA expression pro?le. Tumour Biol 2015; 36:6973e83.
doi: 10.1007/s13277-015-3415-1
[16]
Francis JC, Thomsen MK, Taketo MM, Swain A. b-catenin is required for prostate development and cooperates with Pten loss to drive invasive carcinoma. PLoS Genet 2013; 9: e1003180. https://doi.org/10.1371/journal.pgen.1003180.
doi: 10.1371/journal.pgen.1003180
[17]
White BD, Chien AJ, Dawson DW. Dysregulation of Wnt/beta-catenin signaling in gastrointestinal cancers. Gastroenterology 2012; 142:219e32.
doi: 10.1053/j.gastro.2011.12.001
Tanaka T, Kubota M, Shinohara K, Yasuda K, Kato J-y. In vivo analysis of the cyclin D1 promoter during early embryogenesis in Xenopus. Cell Struct Funct 2003; 28:165e77.
doi: 10.1247/csf.28.165
pmid: 12951437
[20]
Liang S, Mu K, Wang Y, Zhou Z, Zhang J, Sheng Y, et al. CyclinD1, a prominent prognostic marker for endometrial diseases. Diagn Pathol 2013; 8:138. https://doi.org/10.1186/ 1746-1596-8-138.
doi: 10.1186/1746-1596-8-138
pmid: 23947899
[21]
Wang H, Wang H, Zhou X, Makki MS, Wang J. Overexpression of b-βcatenin and cyclin D1 predicts a poor prognosis in ovarian serous carcinomas. Int J Clin Exp Pathol 2014;7: 264e71.
[22]
Livak K, Schmittgen T. Analysis of relative gene expression data using real-time quantitative PCR and the 2-DDCT Method. Methods 2000; 25:402e8.
doi: 10.1006/meth.2001.1262
[23]
Jeong JH, Bhatia A, Toth Z, Oh S, Inn KS, Liao CP, et al. TPL2/COT/MAP3K 8 (TPL2) activation promotes androgen depletion-independent (ADI) prostate cancer growth. PLoS One 2011;6:e16205. https://doi.org/10.1371/journal.pone.0016205.
[24]
Yang C, Wu D, Jia J, Liu D, Li Z, Zhang C, et al. 1 DLC1 as a regulator of proliferation, invasion, cell cycle, and apoptosis in cutaneous squamous cell carcinoma. Tumour Biol 2013; 34:2633e43.
doi: 10.1007/s13277-013-0813-0
[25]
Feng M, Huang B, Du Z, Xu X, Chen Z. DLC-1 as a modulator of proliferation, apoptosis and migration in Burkitt’s lymphoma cells. Mol Biol Rep 2011; 38:1915e20.
doi: 10.1007/s11033-010-0311-z
[26]
Wang C, Wang J, Liu H, Fu Z. Tumor suppressor DLC-1 induces apoptosis and inhibits the growth and invasion of colon cancer cells through the Wnt/beta-catenin signaling pathway. Oncol Rep 2014; 31:2270e8.
doi: 10.3892/or.2014.3057
[27]
Guerra FS, Oliveira RG, Fraga CAM, Mermelstein CDS, Fernandes PD. ROCK inhibition with Fasudil induces beta-catenin nuclear translocation and inhibits cell migra-tion of MDA-MB 231 human breast cancer cells. Sci Rep 2017; 7:13723. https://doi.org/10.1038/s41598-017-14216-z.
doi: 10.1038/s41598-017-14216-z
[28]
Song GL, Jin CC, Zhao W, Tang Y, Wang YL, Li M, et al. Regulation of the RhoA/ROCK/AKT/b-catenin pathway by arginine-speci?c ADP-ribosytransferases 1 promotes migration and epithelial-mesenchymal transition in colon carcinoma. Int J Oncol 2016; 49:646e56.
doi: 10.3892/ijo.2016.3539
[29]
Zhang T, Zheng J, Jiang N, Wang G, Liu C, et al. Overexpression of DLC-1 induces cell apoptosis and prolifer-ation inhibition in the renal cell carcinoma. Cancer Lett 2009; 283:59e67.
doi: 10.1016/j.canlet.2009.03.025
pmid: 19380190
