人内皮抑素对卵巢癌细胞A2780体内外生长的影响及机制

目的 探讨人内皮抑素(human endostatin,hES)对人卵巢癌细胞株A2780体内、外生长的影响,并探讨其可能机制.方法 高保真PCR从含有人内皮抑素基因的人肝脏组织中扩增出人内皮抑素编码区,将其克隆人pcDNA3.1载体中,构建pcDNA3.1-ES重组质粒;将此重组质粒转染人卵巢癌细胞A2780,观察A2780细胞生长.20只BalB/C-nu 品系裸鼠随机分为2组:hES转染组(10只)、对照组(10只),建立人卵巢癌裸鼠模型.1个月后,测量瘤体积,计算抑瘤率;标本行苏木精-伊红染色分析组织学形态;同时用Western blot检测hES、凋亡抑制蛋白Bcl-2的表达;流式细胞仪检测转染对裸鼠移植瘤组织细胞凋亡率影响.结果 hES转染A2780细胞后,细胞生长速率减慢.转染hES的裸鼠肿瘤生长较对照组明显减慢(P<0.01),抑瘤率为74%;转染组肿瘤组织较对照组的肿瘤坏死有所增多,胞核嗜碱性染色明显低于对照组;转染组hES蛋白水平明显高于对照组,而Bcl-2蛋白水平明显低于对照组;流式结果 显示转染使肿瘤细胞的凋亡显著增加(P<0.05).结论 转染hES可抑制卵巢癌A2780细胞体内、外的生长,其机制可能与促进细胞凋亡有关.

The Effects of Human Endostatin on the Growth of Ovarian Cancer Cells A2780 in vitro and in vivo and the Possible Mechanism

Objective To investigate the effects of human endostatin(hES)on ovarian cancer cells A2780 growth in vitro and in vivo and the possible mechanism.Methods The coding region for hES was amplified by PCR from human liver tissue and then subcloned into pcDNA3.1 vector.The growth rate of A2780 cells transfected with hES was observed.A2780 cells(1× 10~7/ml)were inoculated subcutaneously into 20 nude mice,and the mice were randomly divided into two groups:the hES-trans-fected group(group A,n=10)and the control group(group B,n=10).One month later,the size of subcutaneous tumor was measured,and the tumor inhibition rate was calculated.The specimens were stained with HE for the histological analysis.Cell apoptosis in ovarian neoplasm transplantation tissues was determined by flow cytometry.The expression of hES and Bcl-2 was detected by Western blot.Results The growth rate of the hES-transfected ovarian cancer cells A2780 was significantly decreased.In group A,growth of tumor in nude mice was significantly slowed as compared with group B(P<0.01),with the tumor inhibition rate being 74% in group A.The tumor necrosis was increased and the basophilia stain of nucleus decreased significantly in group A as compared with group B.The expression of hES was increased significantly,but that of Bcl-2 was decreased significantly in group A.Flow cytometry revealed that hES transfection could significantly incyease the apoptosis rate of tumor cells(P<0.05).Conclusion Transfection of hES could suppress the growth of ovarian cancer cells A2780 in vitro and in vivo,which may be related tO the promotion of apoptosis.