HSPG和无HS-GAGs链HSPG体外抗乳腺癌的研究

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MDA-MB-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。

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

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 heparitinasel. 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.