RASSF5 inhibits growth and invasion and induces apoptosis in osteosarcoma cells through activation of MST1/LATS1 signaling

Zhou,Xu-Hui; Yang,Chao-Qun; Zhang,Cheng-Lin; Gao,Yang; Yuan,Hong-Bin; Wang,Ce

doi:10.3892/or.2014.3387

RASSF5 inhibits growth and invasion and induces apoptosis in osteosarcoma cells through activation of MST1/LATS1 signaling

Authors:
- Xu-Hui Zhou
- Chao-Qun Yang
- Cheng-Lin Zhang
- Yang Gao
- Hong-Bin Yuan
- Ce Wang
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Affiliations: Department of Orthopedic Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China, Department of Anesthesiology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, P.R. China
Published online on: August 7, 2014 https://doi.org/10.3892/or.2014.3387
Pages: 1505-1512

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Abstract

Ras association (RalGDS/AF-6) domain family member RASSF5 has been implicated in a variety of key biological processes, including cell proliferation, cell cycle regulation and apoptosis. It is believed to play an important role in tumorigenesis as a tumor suppressor in a number of malignancies. Yet, little is known concerning the function and underlying mechanisms of RASSF5 in human osteosarcoma (OS). The expression of RASSF5 was examined by immunohistochemical assay using a tissue microarray in 45 cases of OS tissues. A gain-of-function approach was used to observe the effects of lentiviral vector-mediated overexpression of RASSF5 (Lv-RASSF5) on cell growth, invasion and apoptosis, respectively, as indicated by MTT, Transwell and flow cytometry assays, and the expression levels of mammalian sterile 20-like (MST1) kinase, large tumor suppressor 1 (LATS1), proliferating cell nuclear antigen (PCNA), matrix metallopeptidase-9 (MMP-9) and p53 were detected by real-time PCR and western blot assays in OS cells (MG-63 and U-2 OS). The results indicated that the expression of RASSF5 protein was significantly downregulated in OS tissues compared to that in adjacent non-cancerous tissues (ANCT) (40.0 vs. 73.3%, P=0.002), and had a negative correlation with distant metastasis of the tumor (P=0.01). Overexpression of RASSF5 markedly suppressed cell proliferation and invasion, and induced cell apoptosis in the OS cell lines with increased expression of MST1, LATS1 and p53 and decreased expression of PCNA and MMP-9. Taken together, our findings demonstrate that RASSF5 expression is negatively correlated with distant metastasis of OS, and RASSF5 may function as a tumor suppressor in OS cells through activation of the MST1/LATS1 pathway.

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1	Ta HT, Dass CR, Choong PF and Dunstan DE: Osteosarcoma treatment: state of the art. Cancer Metastasis Rev. 28:247–263. 2009. View Article : Google Scholar : PubMed/NCBI
2	Tan ML, Choong PF and Dass CR: Osteosarcoma: conventional treatment vs. gene therapy. Cancer Biol Ther. 8:106–117. 2009. View Article : Google Scholar : PubMed/NCBI
3	Ottaviani G and Jaffe N: The epidemiology of osteosarcoma. Cancer Treat Res. 152:3–13. 2009. View Article : Google Scholar
4	Chan JJ, Flatters D, Rodrigues-Lima F, et al: Comparative analysis of interactions of RASSF1-10. Adv Biol Regul. 53:190–201. 2013. View Article : Google Scholar : PubMed/NCBI
5	Tommasi S, Dammann R, Jin SG, et al: RASSF3 and NORE1: identification and cloning of two human homologues of the putative tumor suppressor gene RASSF1. Oncogene. 21:2713–2720. 2002. View Article : Google Scholar
6	Vos MD, Martinez A, Ellis CA, et al: The pro-apoptotic Ras effector Nore1 may serve as a Ras-regulated tumor suppressor in the lung. J Biol Chem. 278:21938–21943. 2003. View Article : Google Scholar : PubMed/NCBI
7	Chen J, Lui WO, Vos MD, et al: The t(1;3) breakpoint-spanning genes LSAMP and NORE1 are involved in clear cell renal cell carcinomas. Cancer Cell. 4:405–413. 2003.PubMed/NCBI
8	Hesson L, Dallol A, Minna JD, et al: NORE1A, a homologue of RASSF1A tumour suppressor gene is inactivated in human cancers. Oncogene. 22:947–954. 2003. View Article : Google Scholar
9	Djos A, Martinsson T, Kogner P and Carén H: The RASSF gene family members RASSF5, RASSF6 and RASSF7 show frequent DNA methylation in neuroblastoma. Mol Cancer. 11:402012. View Article : Google Scholar : PubMed/NCBI
10	Steinmann K, Sandner A, Schagdarsurengin U and Dammann RH: Frequent promoter hypermethylation of tumor-related genes in head and neck squamous cell carcinoma. Oncol Rep. 22:1519–1526. 2009.PubMed/NCBI
11	Meng W, Huebner A, Shabsigh A, et al: Combined RASSF1A and RASSF2A promoter methylation analysis as diagnostic biomarker for bladder cancer. Mol Biol Int. 2012:701812012.
12	Foukakis T, Au AY, Wallin G, et al: The Ras effector NORE1A is suppressed in follicular thyroid carcinomas with a PAX8-PPARγ fusion. J Clin Endocrinol Metab. 91:1143–1149. 2006.
13	Nakamura N, Carney JA, Jin L, et al: RASSF1A and NORE1A methylation and BRAF^V600E mutations in thyroid tumors. Lab Invest. 85:1065–1075. 2005. View Article : Google Scholar
14	Geli J, Kogner P, Lanner F, et al: Assessment of NORE1A as a putative tumor suppressor in human neuroblastoma. Int J Cancer. 123:389–394. 2008.
15	Hanahan D and Weinberg RA: The hallmarks of cancer: the next generation. Cell. 144:646–674. 2011. View Article : Google Scholar : PubMed/NCBI
16	Vazquez A, Bond EE, Levine AJ and Bond G: The genetics of the p53 pathway, apoptosis and cancer therapy. Nat Rev Drug Discov. 7:979–987. 2008. View Article : Google Scholar : PubMed/NCBI
17	Richter AM, Pfeifer GP and Dammann RH: The RASSF proteins in cancer; from epigenetic silencing to functional characterization. Biochim Biophys Acta. 1796:114–128. 2009.PubMed/NCBI
18	Macheiner D, Heller G, Kappel S, et al: NORE1B, a candidate tumor suppressor, is epigenetically silenced in human hepatocellular carcinoma. J Hepatol. 45:81–89. 2006. View Article : Google Scholar : PubMed/NCBI
19	Calvisi DF, Evert M and Dombrowski F: Pathogenetic and prognostic significance of inactivation of RASSF proteins in human hepatocellular carcinoma. Mol Biol Int. 2012:8498742012. View Article : Google Scholar : PubMed/NCBI
20	Shinmura K, Tao H, Nagura K, et al: Suppression of hydroxyurea-induced centrosome amplification by NORE1A and down-regulation of NORE1A mRNA expression in non-small cell lung carcinoma. Lung Cancer. 71:19–27. 2011. View Article : Google Scholar : PubMed/NCBI
21	Fernandes MS, Carneiro F, Oliveira C and Seruca R: Colorectal cancer and RASSF family - a special emphasis on RASSF1A. Int J Cancer. 132:251–258. 2013. View Article : Google Scholar : PubMed/NCBI
22	Macheiner D, Gauglhofer C, Rodgarkia-Dara C, et al: NORE1B is a putative tumor suppressor in hepatocarcinogenesis and may act via RASSF1A. Cancer Res. 69:235–242. 2009. View Article : Google Scholar : PubMed/NCBI
23	Geli J, Kiss N, Lanner F, et al: The Ras effectors NORE1A and RASSF1A are frequently inactivated in pheochromocytoma and abdominal paraganglioma. Endocr Relat Cancer. 14:125–134. 2007.
24	Lee CK, Lee JH, Lee MG, et al: Epigenetic inactivation of the NORE1 gene correlates with malignant progression of colorectal tumors. BMC Cancer. 10:5772010.PubMed/NCBI
25	Guo C, Tommasi S, Liu L, et al: RASSF1A is part of a complex similar to the Drosophila Hippo/Salvador/Latstumor-suppressor network. Curr Biol. 17:700–705. 2007. View Article : Google Scholar : PubMed/NCBI
26	Zeng Q and Hong W: The emerging role of the hippo pathway in cell contact inhibition, organ size control, and cancer development in mammals. Cancer Cell. 13:188–192. 2008. View Article : Google Scholar : PubMed/NCBI
27	Chan SW, Lim CJ, Chen L, et al: The Hippo pathway in biological control and cancer development. J Cell Physiol. 226:928–939. 2011. View Article : Google Scholar : PubMed/NCBI
28	Avruch J, Praskova M, Ortiz-Vega S, et al: Nore1 and RASSF1 regulation of cell proliferation and of the MST1/2 kinases. Methods Enzymol. 407:290–310. 2006. View Article : Google Scholar : PubMed/NCBI
29	Ehrkamp A, Herrmann C, Stoll R and Heumann R: Ras and rheb signaling in survival and cell death. Cancers. 5:639–661. 2013. View Article : Google Scholar : PubMed/NCBI
30	Ng YK, Lau WS, Lui VW, et al: Full-length Mst1 exhibits growth promoting function in human hepatocellular carcinoma cells. FEBS Lett. 587:496–503. 2013. View Article : Google Scholar : PubMed/NCBI
31	Aoyama Y, Avruch J and Zhang XF: Nore1 inhibits tumor cell growth independent of Ras or the MST1/2 kinases. Oncogene. 23:3426–3433. 2004. View Article : Google Scholar : PubMed/NCBI
32	Naryzhny SN: Proliferating cell nuclear antigen: a proteomics view. Cell Mol Life Sci. 65:3789–3808. 2008. View Article : Google Scholar : PubMed/NCBI
33	Brinckerhoff CE, Rutter JL and Benbow U: Interstitial collagenases as markers of tumor progression. Clin Cancer Res. 6:4823–4830. 2000.PubMed/NCBI
34	Trieb K and Kotz R: Proteins expressed in osteosarcoma and serum levels as prognostic factors. Int J Biochem Cell Biol. 33:11–17. 2001. View Article : Google Scholar : PubMed/NCBI
35	Grigorian M and Lukanidin E: Activator of metastasis in cancer cells, Mst1/S100A4 protein binds to tumor suppressor protein p53. Genetika. 39:900–908. 2003.(In Russian).