摘要
目的:探讨甲基转移酶抑制剂5-氮杂-2'-脱氧胞苷(5-aza-2'-deoxycytidine,5-Aza-dC)对卵巢癌细胞系OVCAR3中FANCF(Fanconi anemia complementation group F)基因表达的影响,及其对化疗药物紫杉醇敏感性的影响,以了解5-Aza-dC的抗肿瘤效应,寻找卵巢癌治疗的新靶点。方法:采用5-Aza-dC处理FANCF基因甲基化的卵巢癌细胞OVCAR3和FANCF基因非甲基化的卵巢癌细胞A2780。应用甲基化特异性PCR(methylation speci-c-PCR,MSP)、RT-PCR和蛋白质印迹法分析这2种细胞经5-Aza-dC处理前后FANCF基因的甲基化状态以及FANCF mRNA和蛋白的表达情况。MTT法检测卵巢癌细胞的增殖情况。FCM检测紫杉醇对卵巢癌细胞凋亡的影响。建立OVCAR3和A2780细胞裸鼠移植瘤模型,观察5-Aza-dC对裸鼠移植瘤生长的影响,并采用免疫组织化学法检测移植瘤中FANCF蛋白的表达情况。结果:体外实验显示,5-Aza-dC处理后,OVCAR3细胞中FANCF基因发生去甲基化,FANCF mRNA和蛋白的表达水平增加,细胞生长缓慢。5-Aza-dC处理组OVCAR3细胞裸鼠移植瘤体积较未处理组明显缩小,质量也明显降低,FANCF蛋白表达增加;而A2780细胞裸鼠移植瘤组中未观察到上述变化。结论:甲基转移酶抑制剂5-Aza-dC可能通过抑制卵巢癌细胞增殖而成为潜在的卵巢癌治疗药物,但同时也会增加紫杉醇的耐药风险。
Objective： To investigate the effect of methyltransferase inhibitor 5-Aza-dC （5-aza-2＇- deoxycytidine） on the expression of FANCF （Fanconi anemia complementation group F） gene in ovarian cancer OVCAR3 cells, and examine its effect on the sensitivity to chemotherapeutic drug paclitaxel in ovarian cancer in vitro, and to explore the antitumor effect of 5-Aza-dC as a new target for treatment of ovarian cancer. Methods： The ovarian cancer OVCAR3 cells （methylated FANCF gene） and A2780 cells （unmethylated FANCF gene） were treated with 5-Aza-dC. The methylation status of FANCF gene and the expression levels of FANCF mRNA and protein in OVCAR3 and A2780 cells before and after the treatment with 5-Aza-dC were detected by MSP （methylation-specific PCR）, RT-PCR and Western blotting, respectively. The proliferation rate of these ovarian cancer cells was detected by M＇IF method, and the apoptosis rate of these cells treated with paclitaxel was measured by FCM （flow cytometry）. The xenografts were induced in nude mice transplanted with OVCAR3 cells or A2780 cells. The effect of 5-Aza-dC on the growth of the xenografts in nude mice was observed. The expression of FANCF protein in the xenografts was detected by immunohistochemistry. Results： DNA promoter methylation disappears in OVCAR3 cells after treatment with 5-Aza-dC, and the expression levels of FANCF mRNA and protein were increased in vitro. The growth of OVCAR3 cells was also slowed down. The volume and the weight of OVCAR3 ovarian cancer xenografts treated with 5-Aza-dC were both significantly decreased as compared with those of the untreated xenografts in nude mice; the expression of FANCF protein in the xenografts was increased after treatment with 5-Aza-dC. These changes were not observed in A2780 ovarian cancer xenografts in nude mice. Conclusion： The methyltransferase inhibitor 5-Aza-dC may become a potential therapeutic drug in the treatment of ovarian cancer through inhibiting the proliferation of ovarian cancer cells, but it
出处
《肿瘤》
CAS
CSCD
北大核心
2012年第6期413-419,共7页
Tumor
基金
重庆市科委自然科学基金资助项目(编号:cstc2011jjA10081)
