Construction of a Golgi protein 73 modified liposome that mediates HSVtk induced liver cancer cell apoptosis

doi:10.3877/cma.j.issn.1674-0785.2017.19.005

Abstract

Abstract:

Objective

To construct a non-viral gene therapy vector (GP73 modified lipoplexes) and to assess its delivery efficiency and impact on apoptosis in hepatoma cell line HepG2 and normal liver cell line HL-7702.

Methods

Lipoplexes were conjugated with human GP73 using water-soluble carbodidimide, a crossing agent. Then, GP73-lipoplexes were used to carry pGenesil-l plasmid containing EGFP gene and transfect hepatoma cell line HepG2 and normal liver cell line HL-7702. Fluorescence microscopy and flow cytometry were used to assess the transfection efficiency. GP73-lipoplexes were then used to carry PBI-SUR-TK plasmid containing the therapeutic gene HSVtk and transfect hepatoma cells and normal hepatocytes. RT-PCR and Western blot were used to detect the expression of HSVtk in cells, and flow cytometry and CCK-8 assay were used to detect apoptosis.

Results

Compared with HL-7702 cells, HepG2 cells had higher transfection efficiency [(9.68±0.57)% vs (26.37±0.76)%], obvious HSVtk gene production, and significant apoptosis.

Conclusion

GP73 modified lipoplexes can be an effective tool to improve the transfection efficiency and therapeutic effects of exogenous genes in hepatoma cells.

Key words: GP73, Lipoplexes, Hepatoma cell, Transfection efficiency, Apoptosis

Fanxiu Meng, Qi Zhang, Zhijian Yao, Yangyang Yuang, Siheng Zeng, Chang Liu, Yongfang Li, Yingmin Zhang, Na Zhao, Jiahui Lu, Zhixuan Zhang, Gaokai Li, Baofeng Yu, Rui Guo, Hailong Wang, Jun Xie, Gaopeng Li, Jun Xu. Construction of a Golgi protein 73 modified liposome that mediates HSVtk induced liver cancer cell apoptosis[J]. Chinese Journal of Clinicians(Electronic Edition), 2017, 11(19): 2273-2279.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhlcyszz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-0785.2017.19.005

https://zhlcyszz.cma-cmc.com.cn/EN/Y2017/V11/I19/2273

Figures/Tables 6

References 26

[1]	Bruix J, Sherman M. Management of hepatocellular carcinoma: an update [J]. Hepatology, 2011, 53: 1020-1022.
[2]	Cheng JW, Lv Y. New progress of non-surgical treatments for hepatocellularcarcinoma [J]. Med Oncol, 2013, 30(1): 381-389.
[3]	Zhong ZR, Zhang ZR, Deng Y, et al. Preparation of a TK/GCV admipistration system mediated by transefrrin modified proationic liposomes [J]. Pharmazie, 2007, 62(7): 522-527.
[4]	Kladney RD, Bulla GA, Guo L, et al. GP73, a novel Golgi-localized protein upregulated by viral infection [J]. Gene, 2000, 249(1-2): 53-65.
[5]	Zhang Y, Xi Y, Fang J, et al. Identification and characterization of monoclonal antibodies against GP73 for use as a potential biomarker in liver cancer screening and diagnosis [J]. Immunoassay Immunochem, 2016, 37(4): 390-406.
[6]	Zhao Y, Wang M, Cui C, et al. Significance of combined tests of serum golgi glycoprotein 73 and other biomarkers in diagnosis of small primary hepatocellular carcinoma [J]. Cancer Biomark, 2015, 15(5): 677-683.
[7]	Mao Y, Yang H, Xu H, et al. Golgi protein 73(GOLPH2) is a valuable serum marker for hepatocellular carcinoma [J]. Gut, 2010, 59(12): 1687-1693.
[8]	Zhou Y, Yin X, Ying J, et al. Golgi protein 73 versus alpha-feto protein as a biomarker for hepatocellular carcinoma: adiagnosticmeta-analysis [J]. BMC Cancer, 2012, 12(1): 17.
[9]	Ardito G, RevellL, D′Alatri L, et al. Revisited anatomy ofThe recurrent laryngeal nerves [J]. Am J Surg, 2004, 187(2): 249-253.
[10]	秦琴，莱智勇，高然朋, 等. 载脂蛋白Apo E修饰脂质体对肝癌细胞的靶向转染 [J]. 中华实验外科杂志, 2014, 31(8): 1709-1711.
[11]	Lai Z, Qin Q, Yu B, et al. Construction of plasmid vector p AFP-HSVtk-IRES2- EGFP and its effect on the cytotoxicity of ganciclovirto hepatocellular carcinoma [J]. Chin Med J (Engl), 2014, 127(12): 2337-2341.
[12]	Pardridge WM. Drug and gene targeting to the brain with molecular trojan horses [J]. Nat Rev Drug Discov, 2002, 1(2): 131-139.
[13]	Pathak PO, Nagarsenker MS, Barhate CR, et al. Cholesterol anchored arabinogalactan for asialoglycoprotein receptor targe-ting: synthesis, characterization, and proof of concept of hepato-specific delivery [J]. Carbohyd Res, 2015, 408: 33-43.
[14]	Daniels TR., Delgado T, Helguera G, et al. The transferrin receptor part Ⅱ: targeted delivery of therapeutic agentsinto cancer cells [J]. Clin Immunol, 2006, 121(2): 159-176.
[15]	Feng D, Song Y, Shi W, et al. Distinguishing folate-receptor-positive cells from folate-receptor-negative cells using a fluorescence off-on nanoprobe [J]. Anal chem, 2013, 85(13): 6530-6535.
[16]	Kim IY, Kang YS, Lee DS, et al. Antitumor activity of EGFR targeted pH - sensitive immunoliposomes encapsulating gemcitabine in A549 xenograft nude mice [J]. J Control Release, 2009, 140(1): 55-60.
[17]	Wei H, Li B, Zhang R, et al. Serum GP73, a marker for evaluating progression in patients with chronic HBV infections [J]. PLoS One. 2013, 8: e53862.
[18]	Hu JS, Wu DW, Liang S, et al. GP73, a resident Golgi glycoprotein, is sensibility and specificity for hepatocellular carcinoma of diagnosis in a hepatitis B- endemic Asian population [J]. Med Oncol, 2010, 27(2): 339-345.
[19]	Bachert C, Fimmel C, Linstedt AD. Endosomal trafficking and proprotein convertase cleavage of cis Golgi protein GP73 produces marker for hepatocellular carcinoma [J]. Traffic, 2007, 8(10): 1415-1423.
[20]	Gu Y, Chen W, Zhao Y, et al. Quantitative analysis of elevated serum Golgi protein-73 expression in patients with liver diseases [J]. Ann Clin Biochem, 2009, 46(1): 38-43.
[21]	Riener MO, Stenner F, Liewen H, et al. Golgi phosphoprotein 2 (GOLPH2) expression in liver tumors and its value as a serum marker in hepatocellular carcinomas [J]. Hepatology, 2009, 49(5): 1602-1609.
[22]	谢红彬，宫钰，彭涛. 高尔基体驻膜糖蛋白GP73启动子克隆 [J]. 安徽农业科学, 2008, 36(24): 10359-10361, 10401.
[23]	Kladney RD, Tollefson AE, Wold WS, et al. Upregulation of the Golgi protein GP73 by adenovirus infection requires the E1A Ct BP interaction domain [J].Virology, 2002, 301(2): 236-246.
[24]	Liu Y, Zhang X, Sun T, et al. Knockdown of Golgi phosphoprotein 2 inhibits hepatocellular carcinoma cell proliferation and motility [J]. Oncotarget, 2016, 16 (7): 21404-21415.
[25]	Zhang F, Gu Y, Li X, et al. Up-regulated Golgi phosphoprotein 2(GOLPH2) expression in lung adenocarcinoma tissue [J]. Clin. Biochem, 2010, 43 (12): 983-991.
[26]	Qu L, Wang Y, Gong L, et al. Suicide gene therapy for hepatocellular carcinoma cells by surviving promoter-driven expression of the herpes simplex virus thymidine kinase gene [J]. Oncol Rep, 2013, 29(4): 1435-1440.

Please choose a citation manager

Content to export