【作者】 刘勇；

【导师】 马丁；

【作者基本信息】 华中科技大学 ， 妇产科， 2010， 硕士

【摘要】 目的目前研究证实,哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)在各种恶性肿瘤发生发展中发挥重要角色,而其特异抑制剂雷帕霉素被认为能够增强化疗药物的细胞毒性。本研究主要评价雷帕霉素联合顺铂对宫颈癌细胞Hela的协同作用。方法Hela细胞经雷帕霉素和(或)顺铂处理后,以MTT法检测细胞的增殖情况。以流式细胞仪检测细胞凋亡。Western blot检测p-4E-BP1和p-S6K1的表达情况。结果雷帕霉素联合顺铂抑制Hela细胞增殖,且随时间延长抑制作用越显著,与单独用药组相比差异有显著性。雷帕霉素可增强顺铂在Hela细胞中引起的凋亡。经雷帕霉素作用后的Hela细胞表达p-4E-BP1和p-S6K1较对照组下降,两者联用明显下降。结论实验结果显示,雷帕霉素和顺铂联用在抑制Hela细胞增殖并诱导其凋亡上具有协同作用。这预示着雷帕霉素和顺铂联用可能是一种合理的靶向治疗宫颈癌的方法。目的有文献报道,PI-103通过同时抑制PI3K和mTOR激酶途径对神经胶质瘤有强大的杀伤作用。本文通过评价PI-103联用顺铂对C13K细胞体外增殖、凋亡及裸鼠移植瘤生长的影响为卵巢癌的靶向治疗寻找新的药物。方法C13K细胞经PI-103和(或)顺铂处理后,以MTT法检测细胞的增殖情况,以流式细胞仪检测细胞凋亡,Western blot检测p-AKT和p-S6K1的表达情况。建立C13K细胞皮下裸鼠移植瘤模型,随机分成对照组、PI-103组、顺铂组、合并组4组,连续用药4周。观察肿瘤生长情况,计算抑瘤率。结果PI-103联合顺铂抑制C13K细胞增殖,且随时间延长抑制作用越显著,抑制效果优于单独用药。PI-103可增强顺铂在C13K细胞中引起的凋亡。经PI-103作用后的C13K细胞表达p-AKT和p-S6K1较对照组下降,PI-103联合顺铂明显抑制裸鼠C13K细胞皮下移植瘤的生长,较顺铂单药组有明显差异性。结论PI-103增强顺铂对CI3K细胞裸鼠移植瘤生长的抑制作用更多还原

【Abstract】 Objective Recent data suggested that the mammalian target of rapamycin (mTOR) play an important role in the tumorigenesis and development of malignant tumors.Rapamycin,a specific inhibitor of mTOR,is known to enhance the cytotoxicity of chemotherapeutic drugs.This study aimed to evaluate the synergistic effect of rapamycin and cisplatin in cervical cancer cell line Hela.Methods By using Hela cells,cell proliferation was assessed by MTT assay after exposure to rapamycin, cisplatin, or both in combination. Apoptosis was evaluated by flow cytometry. Immunoblot analysis was performed to assess expression of phosphorylation of 4E-BP1 and S6 kinase 1.Results Rapamycin and cisplatin inhibited growth in a time-dependent manner in Hela cell lines . Simultaneous exposure of cisplatin in combination with rapamycin resulted in a significant synergistic antiproliferative effect.Rapamycin increased cisplatin-induced apoptosis in Hela cells .Rapamycin inhibited expression of 4E-BP1 and S6 kinase 1 in Hela cells,and combined treatment with cisplatin inhibited expression over that of rapamycin alone.Conclusions The results of the current study provide evidence of a synergistic relation between rapamycin and cisplatin in both inhibition of Hela cells growth and induction of apoptosis. This suggests that rapamycin and cisplatin may be a rational combination of a targeted therapy for cervical cancer. Objective Data suggested that PI-103 had powerful cytotoxic effects due to combinatorial inhibition of mTOR and PI3 kinase in malignant glioma.This study aimed to evaluate the influence of PI-103 on C13K cells proliferation,apoptosis and the growth of C13K cells xenografls in nude mice.Methods By using C13K cells, cell proliferation was assessed by MTT assay after exposure to PI-103, cisplatin, or both in combination. Apoptosis was evaluated by flow cytometry. Immunoblot analysis was performed to assess expression of phosphorylation of AKT and S6 kinase 1 .C13K cells was subcutaneously transplanted in to 16 nude mice,which were randomly divided into control group,PI-103 group,cisplatin group,combined treatment group,After corresponding treatments for 4 consecutive weeks,the tumor inhibition rate was evaluated。Results PI-103 in combination with cisplatin inhibited growth in a time-dependent manner in C13K cell lines .The inhibitory effect of combination therapy was superior to those treated with PI-103 and cisplatin alone.PI-103 increased cisplatin-induced apoptosis in C13K cells. PI-103 inhibited expression of phosphorylation of AKT and S6 kinase 1 in C13K cells.PI-103 plus cisplatin significantly inhibited the proliferation of xenografted C13K cells in nude mice,and there was significant differences between combined group and cisplatin group.Conclusions PI-103 enhanced growth inhibition of transplanted C13K cells by Cisplatin in nude mice.更多还原