结核分枝杆菌Rv2626c蛋白对RAW264.7细胞凋亡的影响

doi:10.16431/j.cnki.1671-7236.2016.04.006

Abstract

Abstract: The assay was aimed to study the effects of Mycobacterium tuberculosis Rv2626c protein on RAW264.7 cell apoptosis.According to the Rv2626c gene sequence of Mycobacterium tuberculosis in GenBank database, Rv2626c primers were designed.The Rv2626c gene was amplified and cloned into the lentiviral vector pLEX-EGFP.After transfection of lentivirus expressing vector with the helper plasmids into HEK-293T cells, lentivirus were packaged and infected into RAW264.7 cells.Then total protein of the infected cells was extracted.Protein levels of Rv2626c and apoptosis levels of RAW264.7 cells were detected using Western blotting and flow cytometry.The results showed that reconstructed lentivirus expressing vector pLEX-EGFP-Rv2626c and the Rv2626c-overexpressing lentivirus Rv2626c-lv were constructed successfully.The overexpressed Rv2626c protein promoted the RAW264.7 cells apoptosis.Overexpression of Mycobacterium tuberculosis Rv2626c protein significantly promoted apoptosis in RAW264.7 cells.

Key words: Mycobacterium tuberculosis; Rv2626c gene; RAW264.7 cells; apoptosis

CLC Number:

Q786

MENG Lu-ping, SHI Meng-ting, BAO Hai-yang, FU Qiang, SHI Hui-jun, WANG Hui-qin, QIAO Jun, ZHANG Hui, CHEN Chuang-fu. Effects of Mycobacterium tuberculosis Rv2626c Protein on RAW264.7 Cell Apoptosis[J]. , 2016, 43(4): 892-898.

References

[1] Comas I,Coscolla M,Luo T,et al.Out-of-Africa migration and Neolithic coexpansion of Mycobacterium tuberculosis with modern humans[J]. Nature Genetics,2013,45(10):1176-1182.
[2] 崔保亮.结核分枝杆菌rv0199基因在耻垢分枝杆菌中的表达及检测[J].中国畜牧兽医,2014,41(7):9-13.
[3] Rodwell T C,Valafar F,Douglas J,et al.Predicting extensively drug-resistant Mycobacterium tuberculosis phenotypes with genetic mutations[J].Journal of Clinical Microbiology,2014,52(3):781-789.
[4] Rosenkrands I,Slayden R A,Crawford J,et al.Hypoxic response of Mycobacterium tuberculosis studied by metabolic labeling and proteome analysis of cellular and extracellular proteins[J].Journal of Bacteriology,2002,184(13):3485-3491.
[5] 张薇,王丽梅,柏银兰,等.结核分枝杆菌Rv2626c基因的表达、纯化和鉴定[J].科学技术与工程,2009,8:2051-2055.
[6] Geluk A,van Meijgaarden K E,Joosten S A,et al.Innovative strategies to identify M.tuberculosis antigens and epitopes using genome-wide analyses[J].Frontiers in Immunology,2014,5:256.
[7] Obozova T,Artem'ev M,Baranovskiǐ P,et al.Correlation between the expression of Rv3286c,Rv2626c,Rv2031c,AND Rv3133c and the tolerance of Mycobacterium bovis BCG to rifampicin and metronidazole in different physiological conditions] [J].Problemy Tuberkuleza Iboleznei Legkikh,2004,2:34-36.
[8] Zhu Z,Zhang D,Wang H,et al.Expression and serological diagnosis of Mycobacterium tuberculosis CFP-10 and Rv2626c proteins[J].Genetics and Molecular Research,2014,13(3):7398.
[9] Dye C,Scheele S,Dolin P,et al.Global burden of tuberculosis:Estimated incidence,prevalence,and mortality by country[J].Jama,1999,282(7):677-686.
[10] Gengenbacher M,Kaufmann S H E. Mycobacterium tuberculosis:Success through dormancy[J].FEMS Microbiology Reviews,2012,36(3):514-532.
[11] Tyagi J S,Das T K,Kinger A K,et al.Tuberculosis DNA fragment contains genes encoding cell division proteins ftsX and ftsE,a basic protein and homologues of PemK and small protein B[J].Gene,1996,177(1):59-67.
[12] 袁伟.结核分枝杆菌AG85B-ESAT6-HSPX融合基因DNA疫苗的试验研究[D].北京:北京协和医学院,2011.
[13] Hu Y,Movahedzadeh F,Stoker N G,et al.Deletion of the Mycobacterium tuberculosis α-crystallin-like hspX gene causes increased bacterial growth in vivo[J].Infection and Immunity,2006,74(2):861-868.
[14] 罗泰来.表达Rv2031-Rv2626c融合蛋白重组耻垢分枝杆菌的生物学和免疫学特性研究[D].北京:第四军医大学,2012.
[15] Bocchino M,Galati D,Sanduzzi A,et al.Role of mycobacteria-induced monocyte/macrophage apoptosis in the pathogenesis of human tuberculosis[J].The International Journal of Tuberculosis and Lung Disease,2005,9(4):375-383.
[16] 姚楠,董江涛,徐芳,等.结核分枝杆菌Hsp16.3与感染巨噬细胞凋亡的相关性研究[J].中国病原生物学杂志,2012,7(8):569-573.
[17] Ashraf S,Saqib M A,Sharif M Z,et al.Evaluation of diagnostic potential of Rv3803c and Rv2626c recombinant antigens in TB endemic country Pakistan[J].Journal of Immunoassay and Immunochemistry,2014,35(2):120-129.
[18] 王慧勤.结核分枝杆菌分泌蛋白Rv2031c和Rv2626c对细胞凋亡影响的初步研究[D].石河子:石河子大学,2014.

Effects of Mycobacterium tuberculosis Rv2626c Protein on RAW264.7 Cell Apoptosis

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	ZHONG Yueyun, YANG Shuzhan, CHEN Feiyue, LU Zhier, LI Bingxin, LI Wanyan, TIAN Yunbo, HUANG Yunmao, XU Danning, CAO Nan. Effects of LPS on Apoptosis of Goose Embryo-derived Hepatocytes ROS/p38/MAPK Signaling Pathway [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2224-2232.
[2]	LI Can, LI Chuchu, ZENG Wei, ZHANG Ning, JI Chunxiao, CHEN Tao. Recombinant Expression and Function Study of RegⅢγ Protein in Porcine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2414-2426.
[3]	GUO Yaru, LI Kexin, WANG Li, LIU Jinyu, SHI Lei, REN Youshe. Effects of Selenium on Lead-Induced Apoptosis of Mouse Leydig Cells in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1404-1411.
[4]	HUANG Xiaoqian, YUAN Junzhao, LIU Jinhe, ZOU Hui, GU Jianhong, YUAN Yan, LIU Xuezhong, LIU Zongping, BIAN Jianchun. Protective Effect of Honokiol on Cadmium-induced Renal Injury in Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1007-1015.
[5]	FU Jingcheng, HE Junhui, TONG Jiang, WU Wei, GUO Shuang, YANG Yanbin, HAN Yingqian, WANG Yueying, LI Heping. Anti-inflammatory and Antioxidant Roles of HO-1 in Macrophages [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 807-816.
[6]	GU Xianzhe, HOU Ning, LYU Xiaoping, GAO Xueli, LIU Chaonan, WU Wenjie, ZHENG Shimin. Influence of Avian Reticuloendotheliosis Virus Infection on Expression of Bcl-2 and C-myc Genes and Apoptosis in Bursa of Fabricius Cells of SPF Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1488-1496.
[7]	WU Yan, PI Jinsong, ZHANG Hao, LIANG Zhenhua, DU Jinping, PAN Ailuan, SHEN Jie, PU Yuejin. Study on the Mechanism of miR-145-4 Regulating Follicle Development in Egg Ducks [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 809-816.
[8]	ZHU Xinying, GUO Shuai, ZHAN Xiaoyan, ZHONG Dengke, ZHENG Jiangping, HUA Zhimin, LI Tianshun. Effects of miR-188 on the Proliferation, Migration and Apoptosis of Mouse Breast Cancer Cells [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 1153-1161.
[9]	ZHANG Ying, LI Xianqiang, LI Yana, WANG Jiaxiang, WU Yan, PI Jinsong, CHEN Lin. Research Progress on Effects of Granulosa Cell Apoptosis on Follicle Atresia in Poultry [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(12): 4725-4733.
[10]	XIE Shanshan, YANG Qin, DENG Xiaoyu, YI Jihai, WANG Zhen, CHEN Chuangfu. Effects of Mitochondrial Fusion Protein 2 Gene Interference and Overexpression on Macrophage Apoptosis Induced by Brucella [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(1): 283-293.
[11]	QIAO Lianjiang, CHEN Chen, ZHANG Ru, CUI Liheng, XU Li, YANG Yanling. Construction of Eukaryotic Expression Vectors and Bioinformatics Analysis of Rhoa and Ptgs2 Genes in RAW264.7 Cells [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2313-2323.
[12]	JIANG Wenkang, LUO Dongzhang, ZHAO Mingming, LUO Huina, FAN Quanbao, WANG Cuilin, RUAN Huimin, ZHU Haiqi, CHEN Shengfeng, WANG Bingyun. Effects of Cat AD-MSCs and Its Conditioned Medium on Apoptosis and Proliferation of CRFK Damaged by Gentamicin [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2644-2652.
[13]	LI Xuan, TONG Junshuo, ZHANG Dachong, WANG Bingyun, CHEN Shengfeng, LIU Canying. Research Advances on miRNA in Regulating the Apoptosis of Follicular Granulosa Cells and Its Mechanism [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4429-4441.
[14]	LI Yuanhui, FU Chunni, LI Pengcheng, PENG Yukai, LI Liuan, ZHANG Jianbin, QIN Shunyi. Effect of Selenium-Enriched Lactic Acid Bacteria on Apoptosis of Chicken Small Intestinal Epithelial Cells Induced by Aflatoxin B₁ [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4702-4709.
[15]	YAO Jiawei, HUANG Yujie, HUANG Lizhen, RUAN Huimin, CHEN Zhisheng, WANG Bingyun, CHEN Shengfen, ZHANG Hui, BAI Yinshan, LIU Canying, JI Huiqin. Protective Effect of Dimethyl α-Ketoglutarate Pretreatment on Oxidative Stress in Canine Adipose-derived Mesenchymal Stem Cells [J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(12): 4735-4743.