|
|
|
|
|
| Knockdown of the Meq gene in Marek’s disease tumor cell line MSB1 might induce cell apoptosis and inhibit cell proliferation and invasion |
ZHAO Chun-fang1, 2, LI Xin1, HAN Bo1, QU Lu-jiang1, LIU Chang-jun3, Jiu Zhou SONG4, YANG Ning1, LIAN Ling1
|
1 National Engineering Laboratory for Animal Breeding, Department of Animal Genetics and Breeding, College of Animal Science and Technology, China Agricultural University, Beijing 100193, P.R.China
2 Department of Animal Science, Anhui Science and Technology University, Chuzhou 233100, P.R.China
3 Division of Avian Infectious Diseases, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, P.R.China
4 Department of Animal & Avian Sciences, University of Maryland, College Park, Maryland 20742, USA |
|
|
|
|
Abstract Marek’s disease (MD), a highly cell-associated and contagious disease of chickens caused by Marek’s disease virus (MDV) can result in neural lesions, immunosuppression and neoplasia in chicken. The Meq gene is an important oncogene in the MDV genome, and it is expressed highly in MD tumor tissues and MD T-lymphoblastoid cell lines. An experiment was conducted to elucidate the role of Meq in MD tumor transformation. RNA interference technology was used to block its expression, and then analyzed the biological effects of Meq knockdown on the MD tumor cell line MSB1. A small interfering RNA with an interference efficiency of 70% (P<0.01) was transfected into MSB1 cells to knock down the expression of Meq gene. The cell proliferation, cycle and apoptosis were detected post-Meq knockdown. The results showed that MSB1 cell proliferation was downregulated remarkably at 48 h (P<0.01), 60 h (P<0.05) and 72 h (P<0.01) post-Meq knockdown. The cell cycle was unaffected (P>0.05). B-cell lymphoma 2 gene (BCL2) was anti-apoptotic and caspase-6 was the effector in the apoptosis pathway. The activity of caspase-6 was upregulated (P<0.05) significantly and BCL2 gene expression was downregulated (P<0.05) significantly post-Meq knockdown, suggesting cell apoptosis might be induced. MSB1 cell migration did not exhibit any obvious change (P>0.05) post-Meq knockdown, but the expression of two genes (matrix metalloproteinase 2 (MMP2) and MMP9) that are correlated closely to cell invasion was downregulated (P<0.05) remarkably post-Meq knockdown. The Meq knockdown might affect the main features of tumorous cells, including proliferation, apoptosis, and invasion, suggesting that the Meq gene might play a crucial role in interfering with lymphomatous cell transformation.
|
|
Received: 13 November 2019
Accepted:
|
| Fund: The work was supported in part by the National Natural Science Foundation of China (31320103905 and U1901206), the Young Scientist Supporting Project, Program for Changjiang Scholars and Innovative Research Team in University (IRT_15R62), the earmarked fund for China Agriculture Research Systems (CARS-41), the National High Technology Development Plan of China (2013AA102501), the Farm Animals Germplasm Resource Bank, the Beijing Key Laboratory for Animal Genetic Improvement, the University Research Project of Anhui Province, China (KJ2020A0081), the Anhui Provincial Natural Science Foundation, China (2008085QC140), the Foundation of Anhui Science and Technology University, China (DKYJ201901), and the Innovation Funds for Undergraduate Students of Anhui Province, China (S201910879019, S202010879109, and S202010879120). |
|
Corresponding Authors:
Correspondence LUO Xu-gang, E-mail: wlysz@263.net; LIAO Xiu-dong, E-mail: liaoxd56@163.com
|
| About author: YANG Yun-feng, E-mail: 604345120@163.com; * These authors contributed equally to this study. |
Cite this article:
ZHAO Chun-fang, LI Xin, HAN Bo, QU Lu-jiang, LIU Chang-jun, Jiu Zhou SONG, YANG Ning, LIAN Ling.
2020.
Knockdown of the Meq gene in Marek’s disease tumor cell line MSB1 might induce cell apoptosis and inhibit cell proliferation and invasion. Journal of Integrative Agriculture, 19(11): 2767-2774.
|
| Allsopp T E, McLuckie J, Kerr L E, Macleod M, Sharkey J, Kelly J S. 2000. Caspase 6 activity initiates caspase 3 activation in cerebellar granule cell apoptosis. Cell Death and Differentiation, 7, 984–993.
Bertzbach L D, van Haarlem D A, Hartle S, Kaufer B B, Jansen C A. 2019a. Marek’s disease virus infection of natural killer cells. Microorganisms, 7, 588.
Bertzbach L D, Harlin O, Hartle S, Fehler F, Vychodil T, Kaufer B B, Kaspers B. 2019b. IFNα and IFNγ impede Marek’s disease progression. Viruses, 11, 1103.
Bertzbach L D, Kheimar A, Ali F A Z, Kaufer B B. 2018. Viral factors involved in Marek’s disease virus (MDV) pathogenesis. Current Clinical Microbiology Reports, 5, 238–244.
Biggs P M. 1997. The leeuwenhoek lecture, 1997. Marek’s disease herpesvirus: Oncogenesis and prevention. Philosophical Transactions of the Royal Society of London (Series B: Biological Sciences), 352, 1951–1962.
Boodhoo N, Gurung A, Sharif S, Behboudi S. 2016. Marek’s disease in chickens: A review with focus on immunology. Veterinary Research, 47, 119.
Brown A C, Baigent S J, Smith L P, Chattoo J P, Petherbridge L J, Hawes P, Allday M J, Nair V. 2006. Interaction of MEQ protein and C-terminal-binding protein is critical for induction of lymphomas by Marek’s disease virus. Proceedings of the National Academy of Sciences of the United States of America, 103, 1687–1692.
Deng X, Li X, Shen Y, Qiu Y, Shi Z, Shao D, Jin Y, Chen H, Ding C, Li L, Chen P, Ma Z. 2010. The Meq oncoprotein of Marek’s disease virus interacts with p53 and inhibits its transcriptional and apoptotic activities. Virology Journal, 7, 348.
Dong D, Gao J, Sun Y, Long Y, Li M, Zhang D, Gong J, Xu L, Li L, Qin S, Ma J, Jin T. 2015. Adenovirus-mediated co-expression of the TRAIL and HN genes inhibits growth and induces apoptosis in Marek’s disease tumor cell line MSB-1. Cancer Cell International, 15, 20.
Feng R, Chen X, Yu Y, Su L, Yu B, Li J, Cai Q, Yan M, Liu B, Zhu Z. 2010. miR-126 functions as a tumour suppressor in human gastric cancer. Cancer Letters, 298, 50–63.
Haertle S, Alzuheir I, Busalt F, Waters V, Kaiser P, Kaufer B B. 2017. Identification of the receptor and cellular ortholog of the Marek’s disease virus (MDV) CXC chemokine. Frontiers in Microbiology, 8, 2543.
Heidari M, Zhang H M, Sharif S. 2008. Marek’s disease virus induces Th-2 activity during cytolytic infection. Viral Immunology, 21, 203–214.
Kung H J, Xia L, Brunovskis P, Li D, Liu J L, Lee L F. 2001. Meq: An MDV-specific bZIP transactivator with transforming properties. Current Topics in Microbiology and Immunology, 255, 245–260.
Levy A M, Gilad O, Xia L, Izumiya Y, Choi J, Tsalenko A, Yakhini Z, Witter R, Lee L, Cardona C J, Kung H J. 2005. Marek’s disease virus Meq transforms chicken cells via the v-Jun transcriptional cascade: A converging transforming pathway for avian oncoviruses. Proceedings of the National Academy of Sciences of the United States of America, 102, 14831–14836.
Li H, Zhu J, He M, Luo Q, Liu F, Chen R. 2018. Marek’s disease virus activates the PI3K/Akt pathway through interaction of its protein Meq with the p85 subunit of PI3K to promote viral replication. Frontiers in Microbiology, 9, 2547.
Li X, Chiang H I, Zhu J, Dowd S E, Zhou H. 2008. Characterization of a newly developed chicken 44K Agilent microarray. BMC Genomics, 9, 1.
Lian X, Bao C, Li X, Zhang X, Chen H, Jung Y S, Qian Y. 2019. Marek’s disease virus disables the ATR-Chk1 pathway by activating STAT3. Journal of Virology, 93, e02290–e02308.
Liu J L, Ye Y, Lee L F, Kung H J. 1998. Transforming potential of the herpesvirus oncoprotein MEQ: Morphological transformation, serum-independent growth, and inhibition of apoptosis. Journal of Virology, 72, 388–395.
Liu J L, Ye Y, Qian Z, Qian Y, Templeton D J, Lee L F, Kung H J. 1999. Functional interactions between herpesvirus oncoprotein MEQ and cell cycle regulator CDK2. Journal of Virology, 73, 4208–4219.
Loison F, Zhu H, Karatepe K, Kasorn A, Liu P, Ye K, Zhou J, Cao S, Gong H, Jenne D E, Remold-O’Donnell E, Xu Y, Luo H R. 2014. Proteinase 3-dependent caspase-3 cleavage modulates neutrophil death and inflammation. The Journal of Clinical Investigation, 124, 4445–4458.
Nair V. 2018. Spotlight on avian pathology: Marek’s disease. Avian Pathology, 47, 440–442.
Neerukonda S N, Katneni U K, Bott M, Golovan S P, Parcells M S. 2018. Induction of the unfolded protein response (UPR) during Marek’s disease virus (MDV) infection. Virology, 522, 1–12.
Padhi A, Parcells M S. 2016. Positive selection drives rapid evolution of the meq oncogene of Marek’s disease virus. PLoS ONE, 11, e0162180.
Subramaniam S, Johnston J, Preeyanon L, Brown C T, Kung H J, Cheng H H. 2013. Integrated analyses of genome-wide DNA occupancy and expression profiling identify key genes and pathways involved in cellular transformation by a Marek’s disease virus oncoprotein, Meq. Journal of Virology, 87, 9016–9029.
Sun J, Hemler M E. 2001. Regulation of MMP-1 and MMP-2 production through CD147/extracellular matrix metalloproteinase inducer interactions. Cancer Research, 61, 2276–2281.
Tabouret E, Boudouresque F, Farina P, Barrie M, Bequet C, Sanson M, Chinot O. 2015. MMP2 and MMP9 as candidate biomarkers to monitor bevacizumab therapy in high-grade glioma. Neuro-Oncology, 17, 1174–1176.
Tai S S, Hearn C, Umthong S, Agafitei O, Cheng H H, Dunn J R, Niikura M. 2017. Expression of Marek’s disease virus oncoprotein Meq during infection in the natural host. Virology, 503, 103–113.
Teng M, Yu Z H, Zhao P, Zhuang G Q, Wu Z X, Dang L, Li H Z, Ma S M, Cui Z Z, Zhang G P, Wu R, Luo J. 2017. Putative roles as oncogene or tumour suppressor of the Mid-clustered microRNAs in Gallid alphaherpesvirus 2 (GaHV2) induced Marek’s disease lymphomagenesis. The Journal of General Virology, 98, 1097–1112.
Xie Q, Anderson A S, Morgan R W. 1996. Marek’s disease virus (MDV) ICP4, pp38, and meq genes are involved in the maintenance of transformation of MDCC-MSB1 MDV-transformed lymphoblastoid cells. Journal of Virology, 70, 1125–1131.
Yao Y, Nair V. 2014. Role of virus-encoded microRNAs in avian viral diseases. Viruses, 6, 1379–1394.
Yao Y, Zhao Y, Xu H, Smith L P, Lawrie C H, Watson M, Nair V. 2008. MicroRNA profile of Marek’s disease virus-transformed T-cell line MSB-1: Predominance of virus-encoded microRNAs. Journal of Virology, 82, 4007–4015.
Zhang Y, Luo J, Tang N, Teng M, Reddy V, Moffat K, Shen Z, Nair V, Yao Y. 2019. Targeted editing of the pp38 gene in Marek’s disease virus-transformed cell lines using CRISPR/Cas9 system. Viruses, 11, 391.
|
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
