沉默ATF5对卵巢癌裸鼠成瘤的影响  

EFFECT OF ATF5 ON OVARIAN CARCINOMA IN NUDE MICE

作　　者：谷雪[1] 丁朝霞[1] 胡明[2] 王斌[2] 陈爱平[1] 钱冬萌[2] 

机构地区：[1]青岛大学医学院附属医院妇科,山东青岛2660031 [2]青岛大学医学院微生物学教研室,山东青岛2660031

出　　处：《齐鲁医学杂志》2014年第5期377-380,共4页Medical Journal of Qilu

基　　金：山东省高等学校科技计划项目(J12LL11)

摘　　要：目的研究沉默人转录激活因子5(ATF5)后对卵巢癌SKOV-3细胞体外增殖、凋亡及体内成瘤的影响,探讨人类卵巢癌基因治疗的新途径。方法将慢病毒携带的小干扰RNA感染卵巢癌SKOV-3细胞(实验组),空载体转染的卵巢癌SKOV-3细胞(空载体对照组)和卵巢癌SKOV-3细胞(空白对照组)进行对照,对比观察细胞的生长状态;MTT法检测细胞的增殖生长情况;罗氏凋亡检测试剂盒检测细胞凋亡情况;分别于裸鼠皮下接种3组卵巢癌SKOV-3细胞,观察各组裸鼠的存活状态、种植瘤生长情况;RT-PCR检测瘤组织中ATF5mRNA的表达;Western-Blot方法检测瘤组织中ATF5蛋白的表达;苏木素-伊红染色(HE染色)观察瘤组织生物学形态变化。结果经转染ATF5基因的小干扰RNA后,在相同培养时间内3组细胞的增殖和生物学形态没有明显差异,但实验组细胞凋亡数量较其他两组明显增多。皮下接种成瘤后,3组裸鼠的成瘤率没有明显区别,实验组与空载体对照组和空白对照组相比,肿瘤成瘤时间晚,生长缓慢;移植瘤ATF5mRNA表达及蛋白表达降低;细胞分裂象明显减少,凋亡现象明显。结论沉默ATF5对人上皮性卵巢癌裸鼠移植瘤生长有抑制作用,这可能成为卵巢癌基因治疗的新靶点。Objective To study the effect of silencing ATF5 on in-vitro proliferation and apoptosis of ovarian cancer SKOV-3cells and explore a new way of gene therapy for this malignancy. Methods The lentivirus carried with RNA interference was infected with ovarian cancer SKOV-3cells,and compared with an empty vector transfection SKOV-3cells and SKOV-3cells.The growth of the cells was detected by MTT;and the apoptosis by Cell Death Detection Kit.Three kinds of SKOV-3cells were respectively inoculated subcutaneously in nude mice to observe the mouse survival in each group and the growth of implanted tumors.The expressions of ATF5 mRNA and ATF protein in tumor tissue were respectively detected by RT-PCR and Western Blot,HE staining was used to observe the morphologic changes of the tumor tissue. Results After transfection with ATF5 mRNA,the differences of cell proliferation and biologic modality between the three groups-with same cultivation time-were not significant,while the amount of apoptosis was much more in the experimental group than the other two.After inoculation,there was no significant difference between the three groups versus the control and the specificity transfection groups.The expressions of both ATF5 mRNA and ATF5 protein in transplanted tumors declined,cell division decreased,and apoptosis obvious. Conclusion Silence ATF5 shows an inhibition on the growth of transplantation tumor of human epithelial ovarian cancer in nude mice,which is likely to become a new target in gene therapy of ovarian cancer.

关 键 词：转录激活因子类 卵巢肿瘤 RNA干扰 

分 类 号：R737.31[医药卫生—肿瘤]