PYGB siRNA inhibits the cell proliferation of human osteosarcoma cell lines

Zhang,Shuwei; Zhou,Yichi; Zha,Yuanyu; Yang,Yang; Wang,Linlong; Li,Jingfeng; Jin,Wei

doi:10.3892/mmr.2018.9022

PYGB siRNA inhibits the cell proliferation of human osteosarcoma cell lines

Authors:
- Shuwei Zhang
- Yichi Zhou
- Yuanyu Zha
- Yang Yang
- Linlong Wang
- Jingfeng Li
- Wei Jin
View Affiliations

Affiliations: Department of Orthopedics, Zhongnan Hospital of Wuhan University, Wuhan, Hubei 430071, P.R. China
Published online on: May 16, 2018 https://doi.org/10.3892/mmr.2018.9022
Pages: 715-722
Copyright: © Zhang 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

Osteosarcoma is the most common malignant bone carcinoma that primarily occurs between childhood to adolescence. It was suggested by recent research that the Brain type glycogen phosphorylase (PYGB) gene may serve an important role in various types of cancer. In the present study, the PYGB gene was knocked down in order to evaluate the cell viability, invasion and migration of the human osteosarcoma cell lines MG63 and HOS. The expression levels of PYGB in osteosarcoma and bone cyst tissue samples, as well as in the osteosarcoma cell lines were identified using reverse transcription‑quantitative polymerase chain reaction and western blot assay. Subsequently, a Cell Counting kit 8 assay was employed to evaluate cell proliferation. Cell apoptosis rate and cell cycle distribution were measured by flow cytometry. In addition, cell invasion and migration were evaluated through a Transwell assay. The expression levels of the cell apoptosis and tumor metastasis associated proteins B‑cell lymphoma 2 (Bcl‑2), Bcl‑2‑associated X protein, E‑cadherin, Twist, matrix metalloproteinase (MMP)‑9 and MMP2 were measured via western blotting. PYGB exhibited a higher expression level in the osteosarcoma tissue samples, particularly in the human osteosarcoma cell lines MG63 and HOS. Knockdown of PYGB resulted in a decline in cell proliferation, invasion and migration, which was coupled with induced cell apoptosis and cell cycle arrest in MG63 and HOS cells. Furthermore, alterations in the expression of apoptosis and metastasis associated proteins indicated that small interfering (si)PYGB may have regulated cell viability by targeting the Bcl/Caspase and cyclin dependent kinase (CDK)‑1 signaling pathway. In conclusion, PYGB siRNA exerted an inhibitory effect on the cell viability of the human osteosarcoma cells MG63 and HOS by blocking the Caspase/Bcl and CDK1 signaling pathway, highlighting novel potential therapeutic methods for treating osteosarcoma.

View Figures

View References

1	Anoop T, Geetha N, Babanrao SA and Jayasree K: Primary osteosarcoma of rib mimicking lung mass with secondary aneurysmal bone cyst formation. J Thorac Oncol. 9:738–739. 2014. View Article : Google Scholar : PubMed/NCBI
2	Do T, Renker EK and Weber MA: Simulation of teleangiectactic osteosarcoma by aneurysmatic bone cyst. Orthopade. 42:1067–1070. 2013.(In German). View Article : Google Scholar : PubMed/NCBI
3	Janevska V, Spasevska L, Samardziski M, Nikodinovska V, Zhivadinovik J and Trajkovska E: From aneurysmal bone cyst to telangiectatic osteosarcoma with metastasis in inguinal lymph nodes-Case report. Med Pregl. 68:127–132. 2015. View Article : Google Scholar : PubMed/NCBI
4	Aung L, Gorlick RG, Shi W, Thaler H, Shorter NA, Healey JH, Huvos AG and Meyers PA: Second malignant neoplasms in long-term survivors of osteosarcoma. Cancer. 95:1728–1734. 2002. View Article : Google Scholar : PubMed/NCBI
5	Bacci G, Ferrari C, Longhi A, Ferrari S, Forni C, Bacchini P, Palmerini E, Briccoli A, Pignotti E, Balladelli A and Picci P: Second malignant neoplasm in patients with osteosarcoma of the extremities treated with adjuvant and neoadjuvant chemotherapy. J Pediatr Hematol Oncol. 28:774–780. 2006. View Article : Google Scholar : PubMed/NCBI
6	Nagarajan R, Kamruzzaman A, Ness KK, Marchese VG, Sklar C, Mertens A, Yasui Y, Robison LL and Marina N: Twenty years of follow-up of survivors of childhood osteosarcoma: A report from the Childhood Cancer Survivor Study. Cancer. 117:625–634. 2011. View Article : Google Scholar : PubMed/NCBI
7	Jaffe N: Historical perspective of the treatment of osteosarcoma: An interview with Dr Norman Jaffe. Interview by Margaret pearson. J Pediatr Oncol Nurs. 15:90–94. 1998. View Article : Google Scholar : PubMed/NCBI
8	Longhi A, Ferrari S, Tamburini A, Luksch R, Fagioli F, Bacci G and Ferrari C: Late effects of chemotherapy and radiotherapy in osteosarcoma and ewing sarcoma patients: The Italian Sarcoma Group Experience (1983–2006). Cancer. 118:5050–5059. 2012. View Article : Google Scholar : PubMed/NCBI
9	Luetke A, Meyers PA, Lewis I and Juergens H: Osteosarcoma treatment-where do we stand? A state of the art review. Cancer Treat Rev. 40:523–532. 2014. View Article : Google Scholar : PubMed/NCBI
10	Newgard CB, Hwang PK and Fletterick RJ: The family of glycogen phosphorylases: Structure and function. Crit Rev Biochem Mol Biol. 24:69–99. 1989. View Article : Google Scholar : PubMed/NCBI
11	Newgard CB, Littman DR, van Genderen C, Smith M and Fletterick RJ: Human brain glycogen phosphorylase. Cloning, sequence analysis, chromosomal mapping, tissue expression, and comparison with the human liver and muscle isozymes. J Biol Chem. 263:3850–3857. 1988.PubMed/NCBI
12	Philips KB, Kurtoglu M, Leung HJ, Liu H, Gao N, Lehrman MA, Murray TG and Lampidis TJ: Increased sensitivity to glucose starvation correlates with downregulation of glycogen phosphorylase isoform PYGB in tumor cell lines resistant to 2-deoxy-D-glucose. Cancer Chemother Pharmacol. 73:349–361. 2014. View Article : Google Scholar : PubMed/NCBI
13	Shimada S, Maeno M, Akagi M, Hatayama I, Sato T and Sato K: Immunohistochemical detection of glycogen phosphorylase isoenzymes in rat and human tissues. Histochem J. 18:334–338. 1986. View Article : Google Scholar : PubMed/NCBI
14	Barbosa AJ and Castro LP: BGP expression in gastric epithelium and early gastric cancer. Gastric Cancer. 5:123–124. 2002. View Article : Google Scholar : PubMed/NCBI
15	Shimada S, Shiomori K, Honmyo U, Maeno M, Yagi Y and Ogawa M: BGP expression in gastric biopsies may predict the development of new lesions after local treatment for early gastric cancer. Gastric Cancer. 5:130–136. 2002. View Article : Google Scholar : PubMed/NCBI
16	Tashima S, Shimada S, Yamaguchi K, Tsuruta J and Ogawa M: Expression of brain-type glycogen phosphorylase is a potentially novel early biomarker in the carcinogenesis of human colorectal carcinomas. Am J Gastroenterol. 95:255–263. 2000. View Article : Google Scholar : PubMed/NCBI
17	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
18	Kirkin V, Joos S and Zörnig M: The role of Bcl-2 family members in tumorigenesis. Biochim Biophys Acta. 1644:229–249. 2004. View Article : Google Scholar : PubMed/NCBI
19	Manion MK and Hockenbery DM: Targeting Bcl-2 related proteins in cancer therapy. Cancer Biol Ther. 2 4 Suppl 1:S105–S114. 2003. View Article : Google Scholar : PubMed/NCBI
20	Waerner T, Alacakaptan M, Tamir I, Oberauer R, Gal A, Brabletz T, Schreiber M, Jechlinger M and Beug H: ILEI: A cytokine essential for EMT, tumor formation, and late events in metastasis in epithelial cells. Cancer Cell. 10:227–239. 2006. View Article : Google Scholar : PubMed/NCBI
21	Halbleib JM and Nelson WJ: Cadherins in development: Cell adhesion, sorting, and tissue morphogenesis. Genes Dev. 20:3199–3214. 2006. View Article : Google Scholar : PubMed/NCBI
22	Page-McCaw A, Ewald AJ and Werb Z: Matrix metalloproteinases and the regulation of tissue remodelling. Nat Rev Mol Cell Biol. 8:221–233. 2007. View Article : Google Scholar : PubMed/NCBI
23	Thiery JP, Acloque H, Huang RY and Nieto MA: Epithelial-mesenchymal transitions in development and disease. Cell. 139:871–890. 2009. View Article : Google Scholar : PubMed/NCBI
24	Rothberg Gould BE and Bracken MB: E-cadherin immunohistochemical expression as a prognostic factor in infiltrating ductal carcinoma of the breast: A systematic review and meta-analysis. Breast Cancer Res Treat. 100:139–148. 2006. View Article : Google Scholar : PubMed/NCBI
25	Shibata T and Hirohashi S: E-cadherin cell adhesion system in human cancer. Seikagaku. 78:647–656. 2006.(In Japanese). PubMed/NCBI
26	Tycko B, Li CM and Buttyan R: The Wnt/beta-catenin pathway in Wilms tumors and prostate cancers. Curr Mol Med. 7:479–489. 2007. View Article : Google Scholar : PubMed/NCBI
27	Onder TT, Gupta PB, Mani SA, Yang J, Lander ES and Weinberg RA: Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res. 68:3645–3654. 2008. View Article : Google Scholar : PubMed/NCBI
28	Christofori G and Semb H: The role of the cell-adhesion molecule E-cadherin as a tumour-suppressor gene. Trends Biochem Sci. 24:73–76. 1999. View Article : Google Scholar : PubMed/NCBI
29	Leptin M: Twist and snail as positive and negative regulators during Drosophila mesoderm development. Genes Dev. 5:1568–1576. 1991. View Article : Google Scholar : PubMed/NCBI
30	Vesuna F, van Diest P, Chen JH and Raman V: Twist is a transcriptional repressor of E-cadherin gene expression in breast cancer. Biochem Biophys Res Commun. 367:235–241. 2008. View Article : Google Scholar : PubMed/NCBI