HSPG和无HS-GAGs链HSPG体外抗乳腺癌的研究

中国临床药理学与治疗学 ›› 2009, Vol. 14 ›› Issue (6): 617-623.

HSPG和无HS-GAGs链HSPG体外抗乳腺癌的研究

谢强¹, 蒋志文², 祝晓光², 刘浩²

¹安徽省立医院PET-CT 中心, 合肥 230001, 安徽;
²蚌埠医学院药理教研室, 蚌埠 233000, 安徽

收稿日期:2009-01-09 修回日期:2009-06-02 出版日期:2009-06-26 发布日期:2020-10-27
通讯作者: 蒋志文, 男, 教授, 硕导, 研究方向:肿瘤药理学。Tel:0552-3175288 E-mail:zzwwjj51@yahoo.com.cn
作者简介:谢强,男,硕士,研究方向:正电子药物与肿瘤药理学。Tel:0551-2283773 E-mail:jackxie340@21cn.com

Anti-breast cancer experiment research on heparan sulfate proteoglycan and heparan sulfate proteoglycan without heparan sulfate-glycosaminoglycan chain in vitro

XIE Qiang¹, JIANG Zhi-wen², ZHU Xiao-guang², LIU Hao²

¹PET-CT Center of Anhui Provincial Hospital, Hefei 230001, Anhui, China;
²Department of Pharmacology, Bengbu Medical College, Bengbu 233000, Anhui, China

Received:2009-01-09 Revised:2009-06-02 Online:2009-06-26 Published:2020-10-27

摘要/Abstract

摘要： 目的: 比较硫酸乙酰肝素蛋白聚糖(简称HSPG) 和去硫酸乙酰肝素链(简称HS-GAGs 链即HS 链) 的HSPG 的体外抗人乳腺癌细胞MDA-MB-231 的抗增殖和凋亡诱导作用。方法: 对HSPG 进行提取、纯化, 硫酸乙酰肝素酶1 水解HSPG 的HS链;光学显微镜法观察两组分引起的细胞形态学的变化;MTT 比色法检测两组分对肿瘤细胞生长曲线的影响;流式细胞术结合Annexin V-FITC 及PI 双染标记法观察两组分对肿瘤细胞凋亡的影响。结果: HSPG 可致MDA-MB-231 细胞脱落悬浮;MTT 法检测结果显示HSPG 对MDA-MB-231细胞增殖的抑制作用随浓度的增加而增强, 而去HS 链HSPG 只有在高浓度时才呈现出抑制作用。HSPG 能诱导MDA-MB-231 的早期凋亡, 而去HS链HSPG 在高浓度(4 mg/mL) 时对MDA-MB-231 早期凋亡有诱导作用。结论: HSPG 能诱导MDAMB-231 的早期凋亡和抑制细胞生长增殖。作用机制可能与HSPG 的硫酸乙酰肝素链有关。

关键词: 硫酸乙酰肝素蛋白聚糖, 硫酸乙酰肝素链, MDA-MB-231, MCF-7, 凋亡

Abstract: AIM: To compare the antiproliferative effect and apoptosis-inducing activity by HSPG and HSPG without heparan sulfate chains on human mammary cancer cell MDA-MB-231 in vitro. METHODS: HSPG was isolated and purified, the heparan sulfate chains of HSPG was hydrolyzed by heparitinase1.The changes of two components cell morphology were observed by light microscope, the anti-proliferation of HSPG and HSPG without HS chains was measured by MTT assay, the cell growth curve was drawn and the apoptosis was detected by Annexin V PI double staining and flow cytometry. RESULTS: MCF-7 type HSPG induced the exfoliation and suspension of MDA-MB-231 cells.MTT assay showed that different concentrations of HSPG inhibited MDA-MB-231 cells proliferation in a concentration-dependent manner.There was inhibition effect only in high concentrations of HSPG without HS chains.HSPG induced MDA-MB-231 cells'apoptosis at the early stage, however, 4 mg/mL HSPG without HS chains had some effects on the apoptosis at the early stage. CONCLUSION: HSPG could induce MDA-MB-231 cells' apoptosis at the early stage and distinctly inhibit the growth of MDA-MB-231 cells, the mechanism may be related to HS chains of HSPG.

Key words: heparan sulphate proteoglycan, heparan sulphate chain, MDA-MB-231, MCF-7, apoptosis

中图分类号:

R965.2

谢强, 蒋志文, 祝晓光, 刘浩. HSPG和无HS-GAGs链HSPG体外抗乳腺癌的研究[J]. 中国临床药理学与治疗学, 2009, 14(6): 617-623.

XIE Qiang, JIANG Zhi-wen, ZHU Xiao-guang, LIU Hao. Anti-breast cancer experiment research on heparan sulfate proteoglycan and heparan sulfate proteoglycan without heparan sulfate-glycosaminoglycan chain in vitro[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2009, 14(6): 617-623.

参考文献

[1] Eccles SA.Heparanase:breaking down barriers in tumors [J].Nat Med, 1999, 5(7):735-736.
[2] Iozzo RV, San Antonio JD.Heparan sulfate proteoglycans: heavy hitters in the angiogenesis arena[J].J Clin Invest, 2001, 108(3):349.
[3] Mongiat M, Sweeney SM, San Antonio JD, et al.Endorepellin, a novel inhibitor of angiogenesis derived from the C terminus of perlecan[J].J Biol Chem, 2003, 278 (6):4238-4249.
[4] Parish CR, Freeman C, Hulett MD.Heparanase:a key enzyme involved in cell invasion[J].Biochim Biophys Acta, 2001, 1471(3):M99-108.
[5] Parish CR, Freeman C, Hullett MD.Heparanases:a key enzyme involved in cell invasion [R].Biophys Acta, 2001, 1471(3):99-108.
[6] Bloushtain N, Qimron U, Bar-Ilan A.Membrane-associated hepran sulfate proteoglycans are involved in the recognition of cellular targets by NKp30 and NKp46[J].J Immunol, 2004, 173(4):2392-2401.
[7] 蒋志文, Lebourhis Xuefen, Hubert Hondermarck.肿瘤细胞的进行性增殖和Bip GRP78 的合成[J].中国药理学通报, 2002, 18(1):79-83.
[8] 张均田.现代药理实验方法[M].北京医科大学中国协和医科大学联合出版社, 1998:819-820.
[9] Hiroko H, Koji K.Heparan sulfate proteoglycans mediating the epithelial and mesenchymal interaction[J].Seikagaku, 2001, 73(10):1246-1256.
[10] 蒋志文.乳腺上皮细胞硫酸乙酰肝素蛋白聚糖的抗增殖活性及其变异[D].博士学位论文, 2000:9.
[11] Mali M, Elenius K, Miettinen HM, et al.Inhibition of basic fibroblast growth factor-induced growth promotion by overexpression of syndecan-1[J].J Biol Chem, 1993, 268 (32):24215-24222.
[12] Dhodapkar MV, Abe E, Theus A, et al.Syndecan21 is a multifunctional regulator of myeloma pathobiology:Control of tumor cell survival, growth, and bone cell differentiation[J].Blood, 1998, 91(8):2679-2688.
[13] Gonzalez AD, Kaya M, Shi W, et al.OCI25 GPC3, a glypican encoded by a gene that is mutated in the Simpson-Golabi-Behmelovergrowth syndrome, induces apoptosis in a cell line-specificmanner[J].J Cell Biol, 1998, 141(6): 1407-1414.
[14] Schmidt A, Yoshidall K.The antiproliferative activity of arterial heparan sulfate resides in domains enriched with 2-0-sulfated uronic acid residue[J].J Cell Biol, 1992, 267: (27), 19242-19247.
[15] Perrimon N, BernfieldM.Specificities of heparan sulphate proteoglycans in developmental processes[J].Nature, 2000, 404(6779):725-728.
[16] 姜笃银, 付小兵, 盛志勇.硫酸乙酰肝素糖蛋白的结构-功能多样性与相关修饰酶群作用[J].中国病理生理杂志, 2005, 21(5):1020-1026.
[17] Gilat D, Hershkoviz R, Golakorn I, et al.Moleuclar behavior adapts to context:heparanase functions as an extracellular matrix-degrading enzyme or as a T cell adhesion molecule, depending on the local pH[J].J Exp Med, 1995, 181(5):1929-1934.
[18] Walch ET, Marchetti D.Role of neurot rophins and neurot rophins receptors in the in vitro invasion and heparanase production of human prostate cancer cells[J].Clin Exp Metastasis, 1999, 17(4):307-314.
[19] EI-Assal ON, Yamanoi A, Ono T, et al.The clincopathological signifycance of heparanase and basic fibroblast growth factor expressions in hepatocellular carcinoma[J]. Clin Cancer Res, 2001, 7(5):1299-1305.
[20] Parish CR, Freeman C, Brown KJ, et al.Identification of sulfated oliGosaccharide-based inhibitors of tumor growth and metastasis usingnovel in vitro assays for angiogenesis and heparanase activity[J].Cancer Res, 1999, 59(14): 3433-3441.
[21] Woods A.Syndecans:transmembrane modulators of adhesion and matrix assembly[J].J Clin Invest, 2001, 107 (8):935-941.
[22] Bame KJ.Heparanases:endogly cosidases that degrade heparan sulfate proteoglycans[J].Gly cobiology, 2001, 11 (6):91R-98R.
[23] Vlodavsky I, Eldor A, Haimovitz-Friedman A, et al.Expression of heparanase by platelets and circulating cells of the immune system:possible involvement in diapedesis and extravasation[J].Invasion Metastasis, 1992, 12(2):1122-127.