Knockdown of RAC1 and VASP gene expression inhibits breast cancer cell migration

Tian,Yihao; Xu,Liu; He,Yanqi; Xu,Xiaolong; Li,Kai; Ma,Yanbin; Gao,Yang; Wei,Defei; Wei,Lei

doi:10.3892/ol.2018.8930

Knockdown of RAC1 and VASP gene expression inhibits breast cancer cell migration

Authors:
- Yihao Tian
- Liu Xu
- Yanqi He
- Xiaolong Xu
- Kai Li
- Yanbin Ma
- Yang Gao
- Defei Wei
- Lei Wei
View Affiliations

Affiliations: Department of Pathology and Pathophysiology, Hubei Provincial Key Laboratory of Developmentally Originated Disease, School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei 430071, P.R. China
Published online on: June 8, 2018 https://doi.org/10.3892/ol.2018.8930
Pages: 2151-2160
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

The ability of tumor cells to migrate is biologically fundamental for tumorigenesis, growth, metastasis and invasion. The present study examined the role of Ras‑related C3 botulinum toxin substrate (RAC1) and vasodilator‑stimulated phosphoprotein (VASP) in breast cancer cell migration. According to data in Kaplan, Oncomine and The Cancer Genome Atlas, increased expression levels of RAC1 and VASP in breast cancer are associated with decreased cancer cell differentiation, advanced pathological stage and more aggressive tumor subtypes, while increased VASP mRNA expression levels are positively correlated with a poor prognosis in patients with breast cancer. The short hairpin (sh)RNA technique was employed to knock down the expression of RAC1 or VASP. Stable interference with the expression of RAC1 or VASP using RAC1‑shRNA or VASP‑shRNA, respectively, was established in MCF‑7 breast cancer cells. In RAC1‑shRNA or VASP‑shRNA cells, the protein expression levels of RAC1 or VASP were significantly downregulated compared with control cells. The proliferation and migration rates of the RAC1‑shRNA or VASP‑shRNA cells were significantly lower compared with control cells. It was observed that the protein expression levels of VASP also decreased in RAC1‑shRNA cells compared with control cells. The results revealed that RAC1 and VASP may serve important roles in promoting the migration of MCF‑7 breast cancer cells, and that VASP may among the downstream signaling molecules associated with RAC1.

View Figures

View References

1	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
2	Lopez-Marure R, Contreras PG and Dillon JS: Effects of dehydroepiandrosterone on proliferation, migration, and death of breast cancer cells. Eur J Pharmacol. 660:268–274. 2011. View Article : Google Scholar : PubMed/NCBI
3	Krause M, Dent EW, Bear JE, Loureiro JJ and Gertler FB: Ena/VASP proteins: Regulators of the actin cytoskeleton and cell migration. Annu Rev Cell Dev Biol. 19:541–564. 2003. View Article : Google Scholar : PubMed/NCBI
4	Ali M, Rogers LK and Pitari GM: Serine phosphorylation of vasodilator-stimulated phosphoprotein (VASP) regulates colon cancer cell survival and apoptosis. Life Sci. 123:1–8. 2015. View Article : Google Scholar : PubMed/NCBI
5	Pula G and Krause M: Role of Ena/VASP proteins in homeostasis and disease. Handb Exp Pharmacol. 39–65. 2008. View Article : Google Scholar : PubMed/NCBI
6	Sechi AS and Wehland J: ENA/VASP proteins: Multifunctional regulators of actin cytoskeleton dynamics. Front Biosci. 9:1294–1310. 2004. View Article : Google Scholar : PubMed/NCBI
7	Jayakumar T, Lin KC, Lu WJ, Lin CY, Pitchairaj G, Li JY and Sheu JR: Nobiletin, a citrus flavonoid, activates vasodilator-stimulated phosphoprotein in human platelets through non-cyclic nucleotide-related mechanisms. Int J Mol Med. 39:174–182. 2017. View Article : Google Scholar : PubMed/NCBI
8	Dertsiz L, Ozbilim G, Kayisli Y, Gokhan GA, Demircan A and Kayisli UA: Differential expression of VASP in normal lung tissue and lung adenocarcinomas. Thorax. 60:576–581. 2005. View Article : Google Scholar : PubMed/NCBI
9	Cardama GA, Gonzalez N, Maggio J, Menna PL and Gomez DE: Rho GTPases as therapeutic targets in cancer (Review). Int J Oncol. 51:1025–1034. 2017. View Article : Google Scholar : PubMed/NCBI
10	Orgaz JL, Herraiz C and Sanz-Moreno V: Rho GTPases modulate malignant transformation of tumor cells. Small GTPases. 5:e290192014. View Article : Google Scholar : PubMed/NCBI
11	Kazanietz MG and Caloca MJ: The Rac GTPase in cancer: From old concepts to new paradigms. Cancer Res. 77:5445–5451. 2017. View Article : Google Scholar : PubMed/NCBI
12	Zuzga DS, Pelta-Heller J, Li P, Bombonati A, Waldman SA and Pitari GM: Phosphorylation of vasodilator-stimulated phosphoprotein Ser239 suppresses filopodia and invadopodia in colon cancer. Int J Cancer. 130:2539–2548. 2012. View Article : Google Scholar : PubMed/NCBI
13	Espina C, Céspedes MV, García-Cabezas MA, del Pulgar Gómez MT, Boluda A, Oroz LG, Benitah SA, Cejas P, Nistal M, Mangues R and Lacal JC: A critical role for Rac1 in tumor progression of human colorectal adenocarcinoma cells. Am J Pathol. 172:156–166. 2008. View Article : Google Scholar : PubMed/NCBI
14	Fryer BH, Wang C, Vedantam S, Zhou GL, Jin S, Fletcher L, Simon MC and Field J: cGMP-dependent protein kinase phosphorylates p21-activated kinase (Pak) 1, inhibiting Pak/Nck binding and stimulating Pak/vasodilator-stimulated phosphoprotein association. J Biol Chem. 281:11487–11495. 2006. View Article : Google Scholar : PubMed/NCBI
15	Györffy B, Lanczky A, Eklund AC, Denkert C, Budczies J, Li Q and Szallasi Z: An online survival analysis tool to rapidly assess the effect of 22,277 genes on breast cancer prognosis using microarray data of 1,809 patients. Breast Cancer Res Treat. 123:725–731. 2010. View Article : Google Scholar : PubMed/NCBI
16	Parker JS, Mullins M, Cheang MC, Leung S, Voduc D, Vickery T, Davies S, Fauron C, He X, Hu Z, et al: Supervised risk predictor of breast cancer based on intrinsic subtypes. J Clin Oncol. 27:1160–1167. 2009. View Article : Google Scholar : PubMed/NCBI
17	Ye J, Wang W, Xu L, Duan X, Cheng Y, Xin L, Zhang H, Zhang S, Li T and Liu Y: A retrospective prognostic evaluation analysis using the 8th edition of American Joint Committee on Cancer (AJCC) cancer staging system for luminal A breast cancer. Chin J Cancer Res. 29:351–360. 2017. View Article : Google Scholar : PubMed/NCBI
18	Penault-Llorca F: Comments on the new American Joint Committee on Cancer TNM staging for breast cancer. What's new for the pathologist? Ann Pathol. 23:492–495. 2003.(In French). PubMed/NCBI
19	Aznar S and Lacal JC: Rho signals to cell growth and apoptosis. Cancer Lett. 165:1–10. 2001. View Article : Google Scholar : PubMed/NCBI
20	Bar-Sagi D and Hall A: Ras and Rho GTPases: A family reunion. Cell. 103:227–238. 2000. View Article : Google Scholar : PubMed/NCBI
21	Schaefer A, Reinhard NR and Hordijk PL: Toward understanding RhoGTPase specificity: Structure, function and local activation. Small GTPases. 5:62014. View Article : Google Scholar : PubMed/NCBI
22	Zandvakili I, Lin Y, Morris JC and Zheng Y: Rho GTPases: Anti- or pro-neoplastic targets? Oncogene. 36:3213–3222. 2017. View Article : Google Scholar : PubMed/NCBI
23	Hall A: Rho GTPases and the actin cytoskeleton. Science. 279:509–514. 1998. View Article : Google Scholar : PubMed/NCBI
24	Roux P, Gauthier-Rouviere C, Doucet-Brutin S and Fort P: The small GTPases Cdc42Hs, Rac1 and RhoG delineate Raf-independent pathways that cooperate to transform NIH3T3 cells. Curr Biol. 7:629–637. 1997. View Article : Google Scholar : PubMed/NCBI
25	Cuiyan Z, Jie H, Fang Z, Kezhi Z, Junting W, Susheng S, Xiaoli F, Ning L, Xinhua M, Zhaoli C, et al: Overexpression of RhoE in non-small cell lung cancer (NSCLC) is associated with smoking and correlates with DNA copy number changes. Cancer Biol Ther. 6:335–342. 2007. View Article : Google Scholar : PubMed/NCBI
26	Kamai T, Shirataki H, Nakanishi K, Furuya N, Kambara T, Abe H, Oyama T and Yoshida K: Increased Rac1 activity and Pak1 overexpression are associated with lymphovascular invasion and lymph node metastasis of upper urinary tract cancer. BMC Cancer. 10:1642010. View Article : Google Scholar : PubMed/NCBI
27	Li YL, Shao M and Shi DL: Rac1 signalling coordinates epiboly movement by differential regulation of actin cytoskeleton in zebrafish. Biochem Biophys Res Commun. 490:1059–1065. 2017. View Article : Google Scholar : PubMed/NCBI
28	Lim KB, Bu W, Goh WI, Koh E, Ong SH, Pawson T, Sudhaharan T and Ahmed S: The Cdc42 effector IRSp53 generates filopodia by coupling membrane protrusion with actin dynamics. J Biol Chem. 283:20454–20472. 2008. View Article : Google Scholar : PubMed/NCBI
29	Dawid IB, Breen JJ and Toyama R: LIM domains: Multiple roles as adapters and functional modifiers in protein interactions. Trends Genet. 14:156–162. 1998. View Article : Google Scholar : PubMed/NCBI
30	Edwards DC, Sanders LC, Bokoch GM and Gill GN: Activation of LIM-kinase by Pak1 couples Rac/Cdc42 GTPase signalling to actin cytoskeletal dynamics. Nat Cell Biol. 1:253–259. 1999. View Article : Google Scholar : PubMed/NCBI
31	Zhang YT, Xu LH, Lu Q, Liu KP, Liu PY, Ji F, Liu XM, Ouyang DY and He XH: VASP activation via the Gα13/RhoA/PKA pathway mediates cucurbitacin-B-induced actin aggregation and cofilin-actin rod formation. PLoS One. 9:e935472014. View Article : Google Scholar : PubMed/NCBI
32	Gurzu S, Ciortea D, Ember I and Jung I: The possible role of Mena protein and its splicing-derived variants in embryogenesis, carcinogenesis, and tumor invasion: A systematic review of the literature. Biomed Res Int. 2013:3651922013. View Article : Google Scholar : PubMed/NCBI
33	Wu G, Wei L, Yu A, Zhang M, Qi B, Su K, Hu X and Wang J: Vasodilator-stimulated phosphoprotein regulates osteosarcoma cell migration. Oncol Rep. 26:1609–1615. 2011.PubMed/NCBI
34	Liu K, Li L, Nisson PE, Gruber C, Jessee J and Cohen SN: Reversible tumorigenesis induced by deficiency of vasodilator-stimulated phosphoprotein. Mol Cell Biol. 19:3696–3703. 1999. View Article : Google Scholar : PubMed/NCBI
35	Greenwood AI, Kwon J and Nicholson LK: Isomerase-catalyzed binding of interleukin-1 receptor-associated kinase 1 to the EVH1 domain of vasodilator-stimulated phosphoprotein. Biochemistry. 53:3593–3607. 2014. View Article : Google Scholar : PubMed/NCBI