摘要
目的:探讨卵巢癌组织中VEGF、MMP-9蛋白的表达及贝伐单抗联合顺铂对卵巢癌细胞SKOV3的作用机制。方法:收集41例上皮性卵巢恶性肿瘤组织、30例卵巢良性肿瘤组织、30例正常卵巢组织,采用免疫组化法检测卵巢组织中VEGF、MMP-9蛋白的表达情况。MTT实验检测2.5、5.0、10.0、20.0、40.0 mg/L的贝伐单抗对卵巢癌细胞SKOV3的体外增殖抑制作用;流式细胞仪检测5 mg/L贝伐单抗、3 mg/L顺铂单独及联合使用对卵巢癌细胞凋亡的影响;Western blot法检测5 mg/L贝伐单抗、3 mg/L顺铂单独及联合使用对VEGF下游信号通路中关键蛋白AKT、Bad和MMP-9表达的影响。结果:卵巢癌、卵巢良性肿瘤、正常卵巢组织中VEGF蛋白阳性表达率分别为75.6%、40.0%、6.7%;MMP-9蛋白阳性表达率分别为70.7%、20.0%、3.3%,卵巢癌组与卵巢良性肿瘤组和对照组相比均升高(P均<0.001)。贝伐单抗在体外可浓度依赖性地抑制卵巢癌细胞SKOV3的增殖,其IC50为(8.89±0.27)mg/L。贝伐单抗、顺铂可联合诱导卵巢癌细胞SKOV3凋亡(P<0.001);贝伐单抗联合顺铂处理组与单独药物处理组相比,可减弱磷酸化AKT和MMP-9蛋白的表达,增强促凋亡蛋白Bad的表达。结论:VEGF、MMP-9在卵巢癌组织中呈共同高表达状态;贝伐单抗联合顺铂通过对VEGF下游信号通路的调控以及MMP-9蛋白的影响,发挥协同抗肿瘤效应。
Aim： To detect expressions of VEGF and MMP-9 in ovarian cancer tissue and explore mechanism of bevacizumab plus cisplatin on ovarian cancer SKOV3 cells. Methods： A total of 41 epithelial ovarian malignant tumors,30 cases of benign ovarian tumor,30 cases of normal ovarian tissue from ovariectomy were collected. Immunohistochemistry was used to detect the expressions of VEGF and MMP-9 in ovarian tissue. The effect of bevacizumab（ 2. 5,5. 0,10. 0,20. 0,40. 0 mg/L） on SKOV3 cells was detected by MTT assay. Flow cytometry was used to detect the apoptosis of ovarian cancer cells which were treated with bevacizumab（ 5 mg/L） or cisplatin（ 3 mg/L） alone or in combination,and Western blot was used to detect the expressions of AKT,Bad,and MMP-9,the key downstream proteins of VEGF signal pathway. Results：The positive expression rates of VEGF in ovarian cancer,benign ovarian tumor and normal ovarian tissue were 75. 6%,40. 0% and 6. 7%,respectively（ P 0. 001）,and the positive rates of MMP-9 were 70. 7%,20. 0% and 3. 3%,respectively（ P 0. 001）. Bevacizumab inhibited the proliferation of ovarian cancer cells in a concentration-dependent manner,and the IC50 value of bevacizumab was（ 8. 89 ± 0. 27） mg/L. The interaction between bevacizumab and cisplatin was significant（ P 0. 001）,the combined use of bevacizumab and cisplatin performed best compared to the use alone,and synergistic induction of apoptosis in ovarian cancer cells. The bevacizumab plus cisplatin significantly reduced the expression of phosphorylated AKT and MMP-9 protein,and enhanced the expression of the pro-apoptotic protein Bad when compared with bevacizumab or cisplatin alone. Conclusion： VEGF and MMP-9 are overexpressed in ovarian cancer tissue. Bevacizumab combined with cisplatin exerts a synergistic antitumor effect through the regulation of VEGF signaling pathway and MMP-9 protein.
作者
刘端
李红雨
臧星卉
鱼志琪
孙玮
李玉光
LIU Duan;LI Hongyu;ZANG Xinghui;YU Zhiqi;SUN Wei;LI Yuguang(Department of Gynecology and Obstetrics,the Third Affiliated Hospital,Zhengzhou University,Zhengzhou 45005)
出处
《郑州大学学报（医学版）》
CAS
北大核心
2018年第4期516-520,共5页
Journal of Zhengzhou University(Medical Sciences)
