Knockdown of Ubiquitin-specific Peptidase 39 Inhibited the Growth of Osteosarcoma Cells and Induced Apoptosis in Vitro

Overview

Journal Biol Res

Specialty Biology

Date 2017 Apr 14

PMID 28403900

Citations 21

Authors

Zhihua Gan

Kun Han

Shuchen Lin

Haiyan Hu

Zan Shen

Daliu Min

Affiliations

Soon will be listed here.

Abstract

Background: Ubiquitin specific peptidase 39 (USP39), an essential factor in the assembly of the mature spliceosome complex, has an aberrant expression in several cancer. However, its function and the corresponding mechanism on human osteosarcoma has not been fully explored yet.

Methods: The mRNA and DNA copies of USP39 were increased in osteosarcoma cancer tissues compared with the one in human normal tissues according to datasets from the publicly available Oncomine database. A further western blot analysis also demonstrated an aberrant endogenous expression of USP39 in three different osteosarcoma cells. Then lentivirus-mediated short hairpin RNA (shRNA) was designed to silence USP39 in human osteosarcoma cell line U2OS, which is used to test the impact of USP39-silencing on cellular proliferation, colony formation, cell cycle distribution and apoptosis.

Results: Knockdown of USP39 expression in U2OS cell significantly decreased cell proliferation, impaired colony formation ability. A further analysis indicated suppression of USP39 arrested cell cycle progression at G2/M phase via p21 dependent way. In addition, the results of Annexin V/7-AAD staining suggested the knockdown of USP39 could promote U2OS cell apoptosis through PARP cleavage.

Conclusions: These results uncover the critical role of USP39 in regulating cancer cell mitosis and indicate USP39 is critical for osteosarcoma tumorigenesis.

Citing Articles

The Clinical Prediction Value of the Ubiquitination Model Reflecting the Microenvironment Infiltration and Drug Sensitivity in Breast Cancer.

Ma H, Cao J, Zhang Y, Yang J, Wang X, Yu Y J Cancer. 2025; 16(3):784-801.

PMID: 39781342 PMC: 11705054. DOI: 10.7150/jca.101525.

The Role of Deubiquitinating Enzymes in Primary Bone Cancer.

Colaco J, Suresh B, Kaushal K, Singh V, Ramakrishna S Mol Biotechnol. 2024; .

PMID: 39177860 DOI: 10.1007/s12033-024-01254-y.

The Role of Ubiquitination in Osteosarcoma Development and Therapies.

Mao P, Feng Z, Liu Y, Zhang K, Zhao G, Lei Z Biomolecules. 2024; 14(7).

PMID: 39062505 PMC: 11274928. DOI: 10.3390/biom14070791.

The prognostic value of ubiquitin/ubiquitin-like-related genes along with immune cell infiltration and clinicopathological features in osteosarcoma.

Wen J, Wan L, Chen W, Dong X J Orthop Surg Res. 2024; 19(1):356.

PMID: 38879525 PMC: 11179372. DOI: 10.1186/s13018-024-04781-1.

USP39 interacts with SIRT7 to promote cervical squamous cell carcinoma by modulating autophagy and oxidative stress via FOXM1.

Yu J, Yuan S, Song J, Yu S J Transl Med. 2023; 21(1):807.

PMID: 37957720 PMC: 10641974. DOI: 10.1186/s12967-023-04623-4.

References

Lygerou Z, Christophides G, Seraphin B . A novel genetic screen for snRNP assembly factors in yeast identifies a conserved protein, Sad1p, also required for pre-mRNA splicing. Mol Cell Biol. 1999; 19(3):2008-20. PMC: 83994. DOI: 10.1128/MCB.19.3.2008. View

Makarova O, Makarov E, Luhrmann R . The 65 and 110 kDa SR-related proteins of the U4/U6.U5 tri-snRNP are essential for the assembly of mature spliceosomes. EMBO J. 2001; 20(10):2553-63. PMC: 125249. DOI: 10.1093/emboj/20.10.2553. View

Rhodes D, Yu J, Shanker K, Deshpande N, Varambally R, Ghosh D . ONCOMINE: a cancer microarray database and integrated data-mining platform. Neoplasia. 2004; 6(1):1-6. PMC: 1635162. DOI: 10.1016/s1476-5586(04)80047-2. View

Sampath S, Ohi R, Leismann O, Salic A, Pozniakovski A, Funabiki H . The chromosomal passenger complex is required for chromatin-induced microtubule stabilization and spindle assembly. Cell. 2004; 118(2):187-202. DOI: 10.1016/j.cell.2004.06.026. View

Kim D, Behlke M, Rose S, Chang M, Choi S, Rossi J . Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy. Nat Biotechnol. 2004; 23(2):222-6. DOI: 10.1038/nbt1051. View

Gadea B, Ruderman J . Aurora B is required for mitotic chromatin-induced phosphorylation of Op18/Stathmin. Proc Natl Acad Sci U S A. 2006; 103(12):4493-8. PMC: 1401233. DOI: 10.1073/pnas.0600702103. View

Liu S, Rauhut R, Vornlocher H, Luhrmann R . The network of protein-protein interactions within the human U4/U6.U5 tri-snRNP. RNA. 2006; 12(7):1418-30. PMC: 1484429. DOI: 10.1261/rna.55406. View

Kelly A, Sampath S, Maniar T, Woo E, Chait B, Funabiki H . Chromosomal enrichment and activation of the aurora B pathway are coupled to spatially regulate spindle assembly. Dev Cell. 2007; 12(1):31-43. PMC: 1892535. DOI: 10.1016/j.devcel.2006.11.001. View

Riccardi C, Nicoletti I . Analysis of apoptosis by propidium iodide staining and flow cytometry. Nat Protoc. 2007; 1(3):1458-61. DOI: 10.1038/nprot.2006.238. View

10.

Damron T, Ward W, Stewart A . Osteosarcoma, chondrosarcoma, and Ewing's sarcoma: National Cancer Data Base Report. Clin Orthop Relat Res. 2007; 459:40-7. DOI: 10.1097/BLO.0b013e318059b8c9. View

11.

Zhang X, Lan W, Ems-McClung S, Stukenberg P, Walczak C . Aurora B phosphorylates multiple sites on mitotic centromere-associated kinesin to spatially and temporally regulate its function. Mol Biol Cell. 2007; 18(9):3264-76. PMC: 1951741. DOI: 10.1091/mbc.e07-01-0086. View

12.

Valadkhan S . The spliceosome: caught in a web of shifting interactions. Curr Opin Struct Biol. 2007; 17(3):310-5. DOI: 10.1016/j.sbi.2007.05.001. View

13.

Schmittgen T, Livak K . Analyzing real-time PCR data by the comparative C(T) method. Nat Protoc. 2008; 3(6):1101-8. DOI: 10.1038/nprot.2008.73. View

14.

van Leuken R, Luna-Vargas M, Sixma T, Wolthuis R, Medema R . Usp39 is essential for mitotic spindle checkpoint integrity and controls mRNA-levels of aurora B. Cell Cycle. 2008; 7(17):2710-9. DOI: 10.4161/cc.7.17.6553. View

15.

Malumbres M, Barbacid M . Cell cycle, CDKs and cancer: a changing paradigm. Nat Rev Cancer. 2009; 9(3):153-66. DOI: 10.1038/nrc2602. View

16.

Ottaviani G, Jaffe N . The epidemiology of osteosarcoma. Cancer Treat Res. 2010; 152:3-13. DOI: 10.1007/978-1-4419-0284-9_1. View

17.

Guo W, Zhang Y, Chen T, Wang Y, Xue J, Zhang Y . Efficacy of RNAi targeting of pyruvate kinase M2 combined with cisplatin in a lung cancer model. J Cancer Res Clin Oncol. 2010; 137(1):65-72. DOI: 10.1007/s00432-010-0860-5. View

18.

Xiang T, Li L, Yin X, Yuan C, Tan C, Su X . The ubiquitin peptidase UCHL1 induces G0/G1 cell cycle arrest and apoptosis through stabilizing p53 and is frequently silenced in breast cancer. PLoS One. 2012; 7(1):e29783. PMC: 3261155. DOI: 10.1371/journal.pone.0029783. View

19.

Wang H, Ji X, Liu X, Yao R, Chi J, Liu S . Lentivirus-mediated inhibition of USP39 suppresses the growth of breast cancer cells in vitro. Oncol Rep. 2013; 30(6):2871-7. DOI: 10.3892/or.2013.2798. View

20.

Luetke A, Meyers P, Lewis I, Juergens H . Osteosarcoma treatment - where do we stand? A state of the art review. Cancer Treat Rev. 2013; 40(4):523-32. DOI: 10.1016/j.ctrv.2013.11.006. View