【摘要】 目的:探究槐耳多糖(HP)对子宫内膜癌细胞生物学行为和星形胶质细胞升高基因1(AEG-1)表达的影响。方法:采用不同浓度HP(0～11μg/mL)处理人子宫内膜癌ishikawa细胞24 h,通过检测细胞活力确定HP处理浓度；将ishikawa细胞分为Ct组(正常培养)、HP-L组(1μg/mL HP)、HP-H组(5μg/mL HP)、si-NC组(转染si-NC)、si-AEG-1组(转染si-AEG-1)、HP-H+pcDNA组(转染pcDNA+5μg/mL HP)、HP-H+pcDNA-AEG-1组(转染pcDNA-AEG-1+5μg/mL HP)。CCK-8法、EdU染色测定细胞增殖活性；流式细胞术、Transwell、划痕实验分别检测细胞凋亡、侵袭、迁移；Western blot评估AEG-1、细胞周期蛋白D1(CyclinD1)、基质金属蛋白酶(MMP)-2、MMP-9、Bcl-2相关X蛋白(Bax)、半胱氨酸蛋白酶3(Caspase-3)蛋白表达水平。通过细胞实验分组进行裸鼠体内成瘤试验检测HP及AEG-1对裸鼠肿瘤生长的影响。结果:选取1μg/mL HP、5μg/mL HP分别作为后续处理ishikawa细胞的低、高剂量浓度；与Ct组比较，HP-L组、HP-H组细胞OD450值、EdU阳性率、划痕愈合率、侵袭细胞数、AEG-1、CyclinD1、MMP-2、MMP-9蛋白表达水平、裸鼠肿瘤质量均降低(P<0.05),细胞凋亡率、Bax、Caspase-3蛋白表达水平均升高(P<0.05);与Ct组、si-NC组比较，si-AEG-1组细胞OD450值、EdU阳性率、划痕愈合率、侵袭细胞数、AEG-1、CyclinD1、MMP-2、MMP-9蛋白表达水平、裸鼠肿瘤质量均降低，细胞凋亡率、Bax、Caspase-3蛋白表达水平均升高(P<0.05);与HP-H组、HP-H+pcDNA组比较，HP-H+pcDNA-AEG-1组细胞OD450值、EdU阳性率、划痕愈合率、侵袭细胞数、AEG-1、CyclinD1、MMP-2、MMP-9蛋白表达水平、裸鼠肿瘤质量均升高，细胞凋亡率、Bax、Caspase-3蛋白表达水平均降低(P<0.05)。结论:槐耳多糖可能通过下调AEG-1调控ishikawa细胞生物学行为。更多还原

【Abstract】 Objective:To investigate the impact of Huaier polysaccharide(HP) on the biological behaviors of endometrial cancer cells and the expression of astrocyte elevated gene-1(AEG-1).Methods:Different concentrations of HP(0～11 μg/mL) treated human endometrial cancer ishikawa cells for 24 hours, and determined the HP treatment concentration by detecting the cell viability.Ishikawa cells were divided into the Ct group(normally culture),HP-L group(1 μg/mL HP),HP-H group(5 μg/mL HP),si-NC group(transfected with si-NC),si-AEG-1 group(transfected with si-AEG-1),HP-H+pcDNA group(transfected with pcDNA+5 μg/mL HP),and HP-H+pcDNA-AEG-1 group(transfected with pcDNA-AEG-1+5 μg/mL HP).Cell proliferation was measured by CCK-8 method and EdU staining.Flow cytometry, Transwell and scratch assay were used to detect cell apoptosis, invasion and migration, respectively.The protein expression of AEG-1,Cyclin D1,matrix metalloproteinase(MMP)-2,MMP-9,Bcl-2-associated X protein(Bax),and Caspase-3 were detected by Western blot.The effect of HP and AEG-1 on tumor growth in nude mice was detected through cell experiment grouping and in vivo tumor formation test.Results:Select 1 μg/mL HP,5 μg/mL HP as low and high dose concentration of ishikawa cells for subsequent treatment.Compared with the Ct group, the OD450 value of ishikawa cells, EdU positive rate, scratch healing rate, number of invasive cells, the protein expression of AEG-1,CyclinD1,MMP-2,MMP-9,and the tumor mass in nude mice decreased in the HP-L group and HP-H group(P<0.05),and the apoptosis rate, and the expression of Bax and Caspase-3 protein increased(P<0.05).Compared with Ct group and si-NC group, the OD450 value of ishikawa cells, EdU positive rate, scratch healing rate, number of invasive cells, the protein expression of AEG-1,CyclinD1,MMP-2,MMP-9,and the tumor mass in nude mice decreased in the si-AEG-1 group, and the apoptosis rate, and the expression of Bax and Caspase-3 protein increased(P<0.05).Compared with HP-H group and HP-H+pcDNA group, the OD450 value of ishikawa cells, EdU positive rate, scratch healing rate, number of invasive cells, the protein expression of AEG-1,CyclinD1,MMP-2,MMP-9,and the tumor mass in nude mice increased in the HP-H+pcDNA-AEG-1 group, and the apoptosis rate, and the expression of Bax and Caspase-3 protein decreased(P<0.05).Conclusion:Huaier polysaccharide regulates the biological behavior of ishikawa cells by down-regulating AEG-1.更多还原