Tumor suppressor DLC-1 induces apoptosis and inhibits the growth and invasion of colon cancer cells through the Wnt/β-catenin signaling pathway

Wang,Chunyi; Wang,Jialin; Liu,Hong; Fu,Zhongxue

doi:10.3892/or.2014.3057

Tumor suppressor DLC-1 induces apoptosis and inhibits the growth and invasion of colon cancer cells through the Wnt/β-catenin signaling pathway

Authors:
- Chunyi Wang
- Jialin Wang
- Hong Liu
- Zhongxue Fu
View Affiliations

Affiliations: Department of Gastrointestinal Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: March 5, 2014 https://doi.org/10.3892/or.2014.3057
Pages: 2270-2278

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 molecular mechanism of the deleted in liver cancer-1 (DLC-1) gene in human colon cancer growth and invasion. Recombinant lentiviral vectors encoding the DLC-1 gene were constructed for transfection into the human colon cancer cell line SW480. Real-time quantitative polymerase chain reaction (real-time qPCR) and western blot analysis were employed to evaluate the expression of DLC-1, β-catenin, GSK-3β and c-myc in DLC-1-transfected cells. Moreover, cell proliferation assay, cell colony formation assay, cell cycle analysis, apoptosis analysis and cell migration and invasion assays were performed in order to elucidate the role of DLC-1 in colorectal cancer development and progression. Both real-time qPCR and western blot analyses showed that the DLC-1 gene and protein were overexpressed in the DLC-1-transfected SW480 cells. In addition, the expression of β-catenin and GSK-3β was upregulated and the expression of the c-myc gene was downregulated in the DLC-1-transfected SW480 cells. Furthermore, DLC-1 overexpression inhibited cell proliferation, colony formation, migration and invasion, and induced cell cycle arrest at the G1 phase with subsequent apoptosis. DLC-1 inhibits cell growth and invasion in human colon cancer, functioning as a tumor-suppressor gene, possibly through the regulation of the Wnt/β-catenin signaling pathway.

View Figures

View References

1	Siegel R, Naishadham D and Jemal A: Cancer statistics, 2013. CA Cancer J Clin. 63:11–30. 2013. View Article : Google Scholar
2	Moghimi-Dehkordi B and Safaee A: An overview of colorectal cancer survival rates and prognosis in Asia. World J Gastrointest Oncol. 4:71–75. 2012. View Article : Google Scholar : PubMed/NCBI
3	Arasaradnam RP, Commane DM, Bradburn D and Mathers JC: A review of dietary factors and its influence on DNA methylation in colorectal carcinogenesis. Epigenetics. 3:193–198. 2008. View Article : Google Scholar : PubMed/NCBI
4	Beggs AD and Hodgson SV: The genomics of colorectal cancer: state of the art. Curr Genomics. 9:1–10. 2008. View Article : Google Scholar : PubMed/NCBI
5	Fearon ER: Molecular genetics of colorectal cancer. Ann Rev Pathol. 6:479–507. 2011. View Article : Google Scholar : PubMed/NCBI
6	Worthley DL and Leggett BA: Colorectal cancer: molecular features and clinical opportunities. Clin Biochem Rev. 31:31–38. 2010.PubMed/NCBI
7	Yuan BZ, Miller MJ, Keck CL, Zimonjic DB, Thorgeirsson SS and Popescu NC: Cloning, characterization, and chromosomal localization of a gene frequently deleted in human liver cancer (DLC-1) homologous to rat RhoGAP. Cancer Res. 58:2196–2199. 1998.PubMed/NCBI
8	Nagase T, Kikuno R, Hattori A, Kondo Y, Okumura K and Ohara O: Prediction of the coding sequences of unidentified human genes. XIX The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA Res. 7:347–355. 2012. View Article : Google Scholar
9	Guan M, Zhou X, Soulitzis N, Spandidos DA and Popescu NC: Aberrant methylation and deacetylation of deleted in liver cancer-1 gene in prostate cancer: potential clinical applications. Clin Cancer Res. 12:1412–1419. 2006. View Article : Google Scholar : PubMed/NCBI
10	Durkin ME, Yuan BZ, Zhou X, Zimonjic DB, Lowy DR, Thorgeirsson SS and Popescu NC: DLC-1: a Rho GTPase-activating protein and tumour suppressor. J Cell Mol Med. 11:1185–1207. 2007. View Article : Google Scholar : PubMed/NCBI
11	Feng X, Li C, Liu W, Chen H, Zhou W, Wang L, Zhu B, Yao K, Jiang X and Ren C: DLC-1, a candidate tumor suppressor gene, inhibits the proliferation, migration and tumorigenicity of human nasopharyngeal carcinoma cells. Int J Oncol. 42:1973–1984. 2013.PubMed/NCBI
12	Zhang T, Zheng J, Jiang N, Wang G, Shi Q, Liu C and Lu Y: Overexpression of DLC-1 induces cell apoptosis and proliferation inhibition in the renal cell carcinoma. Cancer Lett. 283:59–67. 2009. View Article : Google Scholar : PubMed/NCBI
13	van Amerongen R and Nusse R: Towards an integrated view of Wnt signaling in development. Development. 136:3205–3214. 2009.PubMed/NCBI
14	Jang KY, Kim YN, Bae JS, Chung MJ, Moon WS, Kang MJ, Lee DG and Park HS: Expression of cyclin D1 is associated with β-catenin expression and correlates with good prognosis in colorectal adenocarcinoma. Transl Oncol. 5:370–378. 2012.
15	Lee HK and Jeong SJ: β-Catenin as a valid molecular target for the development of therapeutic inhibitors. JNBT. 2:29–37. 2005.
16	Pritchard CC and Grady WM: Colorectal cancer molecular biology moves into clinical practice. Gut. 60:116–129. 2011. View Article : Google Scholar : PubMed/NCBI
17	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
18	Sengupta N, Gill KA, MacFie TS, Lai CS, Suraweera N, Mcdonald S and Silver A: Management of colorectal cancer: a role for genetics in prevention and treatment? Pathol Res Pract. 204:469–477. 2008. View Article : Google Scholar : PubMed/NCBI
19	Wu WK, Wang XJ, Cheng AS, Luo MX, Ng SS, To KF, Chan FK, Cho CH, Sung JJ and Yu J: Dysregulation and crosstalk of cellular signaling pathways in colon. Crit Rev Oncol Hematol. 86:251–277. 2013. View Article : Google Scholar : PubMed/NCBI
20	Liao YC and Lo SH: Deleted in liver cancer-1 (DLC-1): a tumor suppressor not just for liver. Int J Biochem Cell Biol. 40:843–847. 2008. View Article : Google Scholar : PubMed/NCBI
21	Wu PP, Jin YL, Shang YF, Jin Z, Wu P and Huang PL: Restoration of DLC1 gene inhibits proliferation and migration of human colon cancer HT29 cells. Ann Clin Lab Sci. 39:263–269. 2009.PubMed/NCBI
22	Ullmannova V and Popescu NC: Expression profile of the tumor suppressor genes DLC-1 and DLC-2 in solid tumors. Int J Oncol. 29:1127–1132. 2006.PubMed/NCBI
23	Wilson PJ, McGlinn E, Marsh A, Evans T, Arnold J, Wright K, Biden K, Young J, Wainwright B, Wicking C and Chenevix-Trench G: Sequence variants of DLC1 in colorectal and ovarian tumours. Hum Mutat. 15:156–165. 2000.
24	Seidler HB, Utsuyama M, Nagaoka S, Takemura T, Kitagawa M and Hirokawa K: Expression level of Wnt signaling components possibly influences the biological behavior of colorectal cancer in different age groups. Exp Mol Pathol. 76:224–233. 2004. View Article : Google Scholar : PubMed/NCBI
25	Doucas H, Garcea G, Neal CP, Manson MM and Berry DP: Changes in the Wnt signalling pathway in gastrointestinal cancers and their prognostic significance. Eur J Cancer. 41:365–379. 2005. View Article : Google Scholar : PubMed/NCBI
26	Zhou X, Thorgeirsson SS and Popescu NC: Restoration of DLC-1 gene expression induces apoptosis and inhibits both cell growth and tumorigenicity in human hepatocellular carcinoma cells. Oncogene. 23:1308–1313. 2004. View Article : Google Scholar : PubMed/NCBI
27	Healy KD, Hodgson L, Kim TY, Shutes A, Maddileti S, Juliano RL, Hahn KM, Harden TK, Bang YJ and Der CJ: DLC-1 suppresses non-small cell lung cancer growth and invasion by RhoGAP-dependent and independent mechanisms. Mol Carcinog. 47:326–337. 2008. View Article : Google Scholar : PubMed/NCBI
28	Jin Y, Tian X, Shang Y and Huang P: Inhibition of DLC-1 gene expression by RNA interference in the colon cancer LoVo cell line. Oncol Rep. 19:669–674. 2008.PubMed/NCBI
29	Kim PJ, Plescia J, Clevers H, Fearon ER and Altieri DC: Survivin and molecular pathogenesis of colorectal cancer. Lancet. 362:205–209. 2003. View Article : Google Scholar : PubMed/NCBI
30	Wu PP, Su Y, Jin Z, Wu P, Xu JJ and Huang PL: Effects of DLC1 gene transfection on HT29 cell cycle distribution: the role of Rho A. Southeast Univ (Med Sci Edi). 28:247–251. 2009.
31	Clevers H and Nusse R: Wnt/β-catenin signaling and disease. Cell. 149:1192–1205. 2012.
32	MacDonald BT, Tamai K and He X: Wnt/β-catenin signaling: components, mechanisms, and diseases. Dev Cell. 7:9–26. 2009.
33	Kimelman D and Xu W: beta-catenin destruction complex: insights and questions from a structural perspective. Oncogene. 25:7482–7491. 2006. View Article : Google Scholar : PubMed/NCBI
34	Kawasaki Y, Sato R and Akiyama T: Mutated APC and Asef are involved in the migration of colorectal tumour cells. Nat Cell Biol. 5:211–215. 2003. View Article : Google Scholar : PubMed/NCBI
35	Miyashiro I, Senda T, Matsumine A, Baeg GH, Kuroda T, Shimano T, Miura S, Noda T, Kobayashi S and Monden M: Subcellular localization of the APC protein: immunoelectron microscopic study of the association of the APC protein with catenin. Oncogene. 11:89–96. 1995.PubMed/NCBI
36	van de Wetering M, Sancho E, Verweij C, de Lau W, Oving I, Hurlstone A, van der Horn K, Batlle E, Coudreuse D, Haramis AP, Tjon-Pon-Fong M, Moerer P, van den Born M, Soete G, Pals S, Eilers M, Medema R and Clevers H: The β-catenin/TCF-4 complex imposes a crypt progenitor phenotype on colorectal cancer cells. Cell. 111:241–250. 2002.
37	Zhang T, Otevrel T, Gao Z, Gao Z, Ehrlich SM, Fields JZ and Boman BM: Evidence that APC regulates survivin expression: a possible mechanism contributing to the stem cell origin of colon cancer. Cancer Res. 61:8664–8667. 2001.