肝素结合表皮生长因子在顺铂耐药卵巢癌中的表达及相关性研究

The expression and relationship of Heparin binding-epidermal growth factor-like growth factor in the cisplatin-resistant ovarian cancer

目的检测肝素结合表皮生长因子(HB-EGF)在顺铂耐药卵巢癌中的表达水平并探讨HB-EGF与卵巢癌对顺铂耐药性的关系。方法①体外实验:蛋白印迹法检测卵巢癌亲本细胞株A2780和顺铂耐药株A2780/CDDP中HB-EGF蛋白的表达。分别予以HB-EGF抑制剂CRM197治疗,四甲基偶氮唑蓝(MTT)比色法检测细胞的增殖能力,酶标仪检测加药前后caspase-3蛋白的活性;②体内实验:建立A2780组和A2780/CDDP组卵巢癌裸鼠移植瘤模型,免疫组化法检测2组移植瘤中HB-EGF蛋白的表达。分别予以CRM197治疗观察移植瘤的生长情况并计算CRM197抑瘤率。结果体内外实验结果表明,与A2780组相比,HB-EGF在A2780/CDDP组中高表达。CRM197可有效地降低A2780/CDDP细胞的增殖能力和caspase-3的表达,抑制A2780/CDDP组裸鼠移植瘤的生长(P<0.001)。结论 HB-EGF在顺铂耐药卵巢癌中高表达,并与卵巢癌对顺铂的耐药性相关。

Objective To detect the heparin binding-epidermal growth factor-like growth factor（HB-EGF） expression in cisplatin-resistant ovarian cancer and to investigate the relationship between HB-EGF and the resistance to cisplatin.Methods The human ovarian carcinoma cell line A2780 and the cisplatin-resistant ovarian carcinoma cell line A2780/CDDP were treated with cross-reacting material 197（CRM197,the HB-EGF inhibitor）. Western blot was used to detect HB-EGF expression in A2780 and A2780/CDDP cells.The effect of CRM197on cell proliferation and apoptosis was examined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphe-nyl-tetrazolium bro-mide（MTT）and caspase-3activity assay respectively.（2）In animal experiment,the A2780 and A2780/CDDP cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability following CRM197 treatment. Immunohistochemistry was used to detect the HB-EGF expression of A2780 and A2780/CDDP groups.Results In experiments in vivo and in vitro,compared with A2780 group,HB-EGF protein was significantly over-expressed in A2780/CDDP group.CRM197 as a HB-EGF-targeted agent potently decelerated cell growth and caspase-3activity in A2780/CDDP cells and suppressed tumor growth in A2780/CDDP xenografts（P＆lt;0.001）.Conclusion HBEGF is over-expressed in cisplatin-resistant ovarian cancer and contributes to chemoresistance to cisplatin.