过表达乙酰肝素酶促进胆囊癌细胞系GBC-SD的增殖和迁移

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (2): 198-202.

过表达乙酰肝素酶促进胆囊癌细胞系GBC-SD的增殖和迁移

刘子祥, 周少波^*, 张子艳

蚌埠医学院第二附属医院普外科, 安徽蚌埠 233000

收稿日期:2019-08-11 修回日期:2019-12-02 出版日期:2020-02-05 发布日期:2020-02-05
通讯作者: ^*zhoushaobodoctor@sina.com
基金资助:
安徽高校自然科学研究项目(KJ2018A0240)

Over-expression of heparanase promotes proliferation and migration of gallbladder cancer cell line GBC-SD

LIU Zi-xiang, ZHOU Shao-bo^*, ZHANG Zi-yan

Department of General Surgery, the Second Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China

Received:2019-08-11 Revised:2019-12-02 Online:2020-02-05 Published:2020-02-05
Contact: ^*zhoushaobodoctor@sina.com

摘要/Abstract

摘要： 目的探讨乙酰肝素酶基因过表达对胆囊癌细胞系(GBC-SD)生物学功能的影响。方法构建乙酰肝素酶过表达质粒载体,通过脂质体法转染至胆囊癌细胞系GBC-SD中。用RT-qPCR和Western blot验证转染效果。CCK-8法、流式细胞计量术、划痕愈合实验检测细胞增殖、凋亡、迁移。Western blot检测syndecan-1、FGF-2、E-cadherin蛋白表达情况。结果 GBC-SD细胞经转染后乙酰肝素酶 mRNA(P＜0.01)和蛋白(P＜0.05)表达水平显著升高。与对照组相比,过表达乙酰肝素酶显著上调GBC-SD细胞体外增殖和迁移能力(P＜0.05),下调凋亡水平(P＜0.01),FGF-2蛋白表达显著升高(P＜0.05),而syndecan-1(P＜0.01)和E-cadherin(P＜0.05)蛋白表达降低。结论乙酰肝素酶过表达促进胆囊癌细胞系GBC-SD的增殖与迁移,其机制可能与FGF-2调节E-cadherin有关。

关键词: 胆囊癌, 乙酰肝素酶, 增殖, 凋亡, 迁移

Abstract: Objective To investigate the effect of heparanase over-expression on the biological function of gallbladder carcinoma cell line GBC-SD. Methods The heparanase over-expression plasmid vector was constructed and transfected into GBC-SD cells by liposome method. The transfection efficiency was verified by RT-qPCR and Western blot. CCK-8 assay, flow cytometry and scratch healing experiment were used to detect the cell proliferation, apoptosis and migration.The expression of syndecan-1, FGF-2 and E-cadherin protein was detected by Western blot. Results The expression of heparanase mRNA(P＜0.01) and protein(P＜0.05) in gallbladder cancer cell line GBC-SD was significantly increased after transfection.Compared with the control group,the over-expression of heparanase significantly up-regulated the proliferation and migration ability of GBC-SD cells in vitro(P＜0.05), down-regulated the level of apoptosis(P＜0.05),and increased the expression of FGF-2 protein, while decreasing the expression of syndecan-1(P＜0.01) and E-cadherin(P＜0.05). Conclusions Over-expression of heparanase promotes the proliferation and migration of gallbladder cancer cell line GBC-SD, and the mechanism may be related to its regulation of E-cadherin through FGF-2.

Key words: gallbladder carcinoma, heparanase, proliferation, apoptosis, migration

中图分类号:

R735.8

刘子祥, 周少波, 张子艳. 过表达乙酰肝素酶促进胆囊癌细胞系GBC-SD的增殖和迁移[J]. 基础医学与临床, 2020, 40(2): 198-202.

LIU Zi-xiang, ZHOU Shao-bo, ZHANG Zi-yan. Over-expression of heparanase promotes proliferation and migration of gallbladder cancer cell line GBC-SD[J]. Basic & Clinical Medicine, 2020, 40(2): 198-202.

参考文献

[1]Hundal R, Shaffer EA. Gallbladder cancer: epidemiology and outcome [J]. Clin Epidemiol, 2014, 6: 99-109.
[2]Siegel RL, Fedewa SA, Miller KD, et al. Cancer statistics for Hispanics/Latinos, 2015 [J]. CA Cancer J Clin, 2015, 65: 457-480.
[3]Masola V, Gambaro G, Tibaldi E, et al. Heparanase and syndecan-1 interplay orchestrates fibroblast growth factor-2-induced epithelial-mesenchymal transition in renal tubular cells [J]. J Biol Chem, 2012, 287: 1478-1488.
[4]Liu LP, Sheng XP, Shuai TK, et al. Helicobacter pylori promotes invasion and metastasis of gastric cancer by enhancing heparanase expression [J]. World J Gastroenterol, 2018, 24: 4565-4577.
[5]Chen X, Jiang W, Yue C, et al. Heparanase contributes to trans-endothelial migration of hepatocellular carcinoma cells [J]. J Cancer, 2017, 8: 3309-3317.
[6]Song JW, Tan YX, Li SB, et al. Gemcitabine-induced heparanase promotes aggressiveness of pancreatic cancer cells via activating EGFR signaling [J]. Oncotarget, 2017, 8: 58417-58429.
[7]Jin H, Zhou S, Yang S, et al. Heparanase overexpression down-regulates syndecan-1 expression in a gallbladder carcinoma cell line [J]. J Int Med Res, 2017, 45: 662-672.
[8]Rivara S, Milazzo FM, Giannini G. Heparanase: a rainbow pharmacological target associated to multiple pathologies including rare diseases [J]. Future Med Chem, 2016, 8: 647-680.
[9]Wang L, Huang X, Kong G, et al. Ulinastatin attenuates pulmonary endothelial glycocalyx damage and inhibits endothelial heparanase activity in LPS-induced ARDS [J]. Biochem Biophys Res Commun, 2016, 478: 669-675.
[10]Vlodavsky I, Miao HQ, Medalion B, et al. Involvement of heparan sulfate and related molecules in sequestration and growth promoting activity of fibroblast growth factor [J]. Cancer Metastasis Rev, 1996, 15: 177-186.
[11]Vlodavsky I, Bar-Shavit R, Ishai-Michaeli R, et al. Extracellular sequestration and release of fibroblast growth factor: a regulatory mechanism [J]. Trends Biochem Sci, 1991, 16: 268-271.
[12]Secchi MF, Masola V, Zaza G, et al. Recent data concerning heparanase: focus on fibrosis, inflammation and cancer [J]. Biomol Concepts, 2015, 6: 415-421.
[13]Strutz F, Zeisberg M, Ziyadeh FN, et al. Role of basic fibroblast growth factor-2 in epithelial-mesenchymal transformation [J]. Kidney Int, 2002, 61: 1714-1728.

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