畸胎瘤细胞源性生长因子反义核酸载体对高度恶性人卵巢癌细胞增殖和侵袭的抑制效应及相关机制  

Inhibitory effects of an antisense PCDGF vector on proliferation and invasion of highly malignant ovarian cancer cells and the related mechanism

作　　者：刘玉兰[1] 王燕[1] 郎雁[1] 吴绪峰[1] 熊俊[1] 朱小红[1] 张幼鸿[1] 张水娟[1] 龚丽艳[1] 卢运萍[2] 马丁[2] 

机构地区：[1]湖北省妇幼保健院妇科,武汉430070 [2]华中科技大学同济医学院附属同济医院妇产科

出　　处：《中华肿瘤杂志》2009年第2期90-94,共5页Chinese Journal of Oncology

基　　金：国家973重大基础科学基金（2002CB513100）;国家杰出青年基金（30025017）

摘　　要：目的观察畸胎瘤细胞源性生长因子（PCDGF）反义核酸载体对高度恶性人卵巢癌细胞株Sw626和A2780增殖和侵袭的抑制效应，并初步探讨其相关机制。方法采用二苯基溴化四氮唑蓝（Mrrr）法和Boyden小室体外侵袭实验，检测PCDGF反义RNA真核表达载体对Sw626和A2780细胞增殖和侵袭能力的影响。采用Westernblot技术，检测转染PCDGF反义RNA真核表达载体前后Sw626细胞cyclin D1和CDK4蛋白表达的变化。采用逆转录聚合酶链反应（RT—PCR）和明胶酶谱法，分析PCDGF反义RNA真核表达载体对Sw626细胞基质金属蛋白酶2（MMP-2）表达和活性的影响。结果与空白对照组相比，PCDGF反义核酸载体转染组Sw626和A2780细胞的增殖抑制率分别为72．9％和70．9％，侵袭能力分别被抑制了62．9％和59．0％。转染组Sw626细胞cyclin D1和CDK4蛋白的表达水平分别为0．38±0．08和0．37±0．13，明湿低于空白对照组（0．84±0．11和0．64±0．11，P〈0．01）。与空白对照组（0．89±0．09）相比，转染组Sw626细胞MMP-2mRNA的表达水平（0．66±0．11）虽未见降低（P〉0．05），但MMP-2酶原的活性被明显抑制。结论PCDGF反义核酸可显著抑制高度恶性人卵巢癌细胞株Sw626和A2780的增殖和侵袭能力，并逆转其部分恶性表型，这可能与其能下调cyclin D1和CDK4蛋白的表达并抑制MMP-2酶原的活性有关；PCDGF可以作为卵巢癌治疗的新靶点。Objective To investigate the inhibitory effects of an antisense PC cell derived growth factor （PCDGF） vector on proliferation and invasion of highly malignant ovarian cancer cell lines Sw626 and 32780 cells, and preliminarily explore the related mechanisms. Methods MTT assay and Boyden chamber in vitro invasion assay were employed to detect the changes of proliferation and invasion ability in the Sw626 and A2780 cells transfected with anti-sense PCDGF. The expression levels of cyclin D1 and CDK4 proteins before and after transfection were detected by Western blotting. The effects on the expression and activity of MMP-2 were evaluated by quantitative RT-PCR and zymography, respectively. Results Comparing with the blank group, the proliferation inhibition rate of the Sw626 and A2780 cells transfected with anti-sense PCDGF was 72.9% and 70.9%, respectively, and the invasion ability was inhibited by 62.9% and 59.0% , respectively. The levels of cyclin D1 and CDK4 protein expression in antisense PCDGF transfected ceils were 0.38 ±0.08 and 0.37±0.13, respectively,all significantly lower than 0.84±0.11 and 0.64 ± 0.11, respectively, in the blank group （ P 〈 0. 01 ）. The MMP-2 mRNA expression level in antisense PCDGF transfected cell group was 0.66 ± 0.11, not significantly decreased in comparison with 0.89 ± 0.09 in the blank group （P 〉 0.05）, but the activity of MMP-2 was inhibited significantly. Conclusion The antisense PCDGF vector may inhibit markedly the proliferation and invasion of highly malignant ovarian cancer cells, and partially reverses their malignant phenotype. It seems to be related with down-regulating the expression of cyclin D1 and CDK4 and inhibiting the activity of MMP-2. Our findings indicate that PCDGF may become a new target for antisense gene therapy of ovarian cancer.

关 键 词：卵巢肿瘤 反义核酸载体 增殖 侵袭 畸胎瘤细胞源性生长因子 

分 类 号：R686[医药卫生—骨科学]