维生素C与宫颈癌HeLa细胞凋亡关系的研究

目的:探讨维生素C是否能诱导实体肿瘤宫颈癌HeLa细胞凋亡,及其作用机制.方法:采用不同浓度的维生素C处理HeLa细胞不同时段,显微镜下观察细胞的形态学变化;噻唑蓝(MTT)法和流式细胞分析法测定细胞生长抑制情况;检测不同浓度的维生素C[(0.25-4)mmol/L]处理细胞15min、30min、1h、2h、5h、8h、12h、24 h不同时段后细胞内过氧化氢(H2O2)水平变化情况.结果:0.25 mmol/L维生素C对HeLa细胞生长无明显影响,而0.5 mmol/L维生素C处理HeLa细胞24 h后细胞的生长受到抑制,表现出细胞凋亡的特征,MTT检测此时细胞的存活率只有59.94%,随着维生素C浓度的增加和作用时间的延长效果更加明显,与流式细胞分析的结果一致.用DCFH-DA标记细胞,检测细胞内H2O2水平,发现不同浓度维生素C处理HeLa细胞15min后,除0.25 mmol/L组外,其他组的细胞内H2O2水平比对照组升高(4mmol/L组为对照组的162.46%),且呈现明显的剂量依赖性,持续到30min达到高峰(4mmol/L组为为对照组的174.55%),1 h后检测细胞内H2O2水平已下降,以后各时间段检测到的细胞内H2O2水平均比对照组低.结论:低浓度维生素C(≤0.25mmol/L)单独处理实体肿瘤宫颈癌HeLa细胞不能抑制其生长,而较高浓度(0.5mmol/L)维生素C能诱导HeLa细胞凋亡,且与细胞内H2O2水平改变有密切关系 .H2O2积累是维生素C诱导肿瘤细胞凋亡途径中的早期事件.     

褪黑素通过下调HIF-1α表达增强卵巢癌细胞OVCAR3顺铂敏感性

目的:探讨褪黑素对卵巢癌细胞顺铂敏感性的影响及其机制。方法:采用不同浓度褪黑素联合顺铂作用于卵巢癌细胞OVCAR3,MTT法检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测凋亡相关蛋白及信号通路蛋白AKT及ERK的磷酸化,同时检测褪黑素对HIF-1α蛋白表达的影响。结果:MTT结果显示,褪黑素和顺铂联合作用能够显著抑制OVCAR3细胞增殖(P<0.05)。流式细胞仪检测结果显示,褪黑素和顺铂联合作用相较于单独顺铂作用或褪黑素作用能够显著增加OVCAR3细胞的凋亡。Western blot结果显示,褪黑素与顺铂联合作用能够增加促凋亡蛋白BAX及Caspase3的表达,抑制抗凋亡蛋白BCL2的表达,检测信号通路结果显示,褪黑素与顺铂联合作用能够促进AKT及ERK的磷酸化,同时褪黑素能够抑制HIF-1α蛋白的表达。结论:褪黑素通过抑制HIF-1α的表达使OVCAR3细胞对顺铂敏感。     

重组人TRAIL蛋白联合顺铂对人卵巢癌细胞生长及凋亡的影响

背景与目的：研究发现肿瘤坏死因子的相关凋亡诱导配体(tumor necrosis factor-related apoptosis inducing ligand,TRAIL)可以增强化疗药物对肿瘤细胞的杀伤作用。本研究旨在探讨TRAIL与顺铂联合应用对体外培养的卵巢癌细胞SKOV3和OVCAR3生长凋亡的影响及可能的诱导机制。方法：利用MTT法和流式细胞仪检测在顺铂和重组人TRAIL蛋白共同作用下,SKOV3和OVCAR3细胞的增殖抑制效应及细胞凋亡程度;并应用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)检测药物处理后TRAIL死亡受体DR4、DR5的mRNA表达水平;同时用蛋白[质]印迹法(Western blot)检测DR4、DR5的蛋白表达水平。结果：SKOV3和OVCAR3细胞均对TRAIL蛋白敏感,随着TRAIL蛋白浓度的升高,细胞的生长抑制率可达64%;而TRAIL与顺铂联合用药对两种细胞的抑制率均达到92%以上,对细胞的增殖抑制呈现高效协同作用,与单独用药组比较差异有统计学意义(P<0.05);TRAIL和顺铂联合组两种细胞凋亡率分别为(31.50±0.79)%和(36.60±1.31)%,显著高于单独用药组;RTFQ-PCR和Western blot检测结果显示,SKOV3和OVCAR3细胞在TRAIL与顺铂联合用药后,死亡受体DR4、DR5表达水平均显著上调。结论：在体外,TRAIL与化疗药物顺铂联用能明显抑制卵巢癌细胞增殖,诱导肿瘤细胞凋亡。TRAIL能明显增强顺铂对卵巢癌细胞的敏感性,其诱导机制可能与死亡受体DR4、DR5表达水平上调有关。     

ING4基因表达上调对卵巢上皮性癌OVCAR细胞顺铂耐药性的影响

目的：探讨ING4基因在表达上调的卵巢上皮性癌（卵巢癌）细胞系OVCAR细胞中对顺铂耐药性的影响。方法：脂质体介导的ING4-Ad及相应阴性对照片段转染人卵巢癌细胞OVCAR,实验分为3组：转染组、阴性对照组、空白对照组。实时荧光定量PCR技术检测3组细胞中ING4基因的表达;采用四甲基偶氮唑蓝（ MTT）法测定各组细胞增殖抑制率和绘制细胞生长曲线,集落形成实验测定细胞生长能力,检测3组细胞对顺铂的敏感性,及不同浓度顺铂和作用不同时间后3组细胞生长的抑制率和凋亡率及细胞周期变化。结果：转染组、阴性对照组及空白对照组OVCAR细胞中ING4的表达量分别为66.76±11.40、1.28±0.09、1.22     

Aurora A调控活性氧对卵巢癌顺铂耐药的影响

背景与目的：极光激酶A（Aurora A kinase,Aurora A）属于丝/苏氨酸蛋白激酶家族中的一员,可促进卵巢癌的化疗抵抗。活性氧（reactive oxygen species,ROS）由机体正常代谢产生,参与细胞信号转导过程。肿瘤细胞由于代谢旺盛及细胞内氧化还原体系的失衡,ROS水平高于正常细胞。并且耐药细胞株中ROS水平降低,提示ROS在肿瘤耐药中发挥作用。该研究旨在探讨ROS在Aurora A介导顺铂化疗耐药中的作用。方法：采用二氯二氢荧光素-乙酰乙酸酯（2’, 7’-dichlorofluorescin diacetate,DCFH-DA）法检测人卵巢癌细胞株A2780和顺铂耐药株A2780/CDDP中ROS的水平,采用四甲基偶氮唑蓝（methyl thiazolyl tetrazolium,MTT）法检测顺铂的半数抑制浓度（50% inhibitory concentration,IC₅₀）;构建Aurora A过表达和沉默细胞株,检测细胞内ROS水平;加入ROS清除剂N-乙酰-L-半胱氨酸（N-acetyl-L-cysteine,NAC）,检测顺铂的IC₅₀;采用蛋白［质］印迹法（Western blot）检测相关信号通路,探讨可能的分子机制。结果：顺铂耐药株中ROS水平低于对照细胞株。加入NAC降低细胞内ROS可增强细胞对顺铂的耐药性。Aurora A过表达,细胞内ROS水平下降,可增强细胞对顺铂的耐药;而Aurora A沉默后,细胞内ROS升高,则可提高细胞对顺铂的敏感性。Aurora A过表达,促进AMP活化蛋白激酶（AMP-activated protein kinase,AMPK）的磷酸化及糖酵解。结论：Aurora A可能通过促进AMPK磷酸化及糖酵解过程,降低细胞内ROS水平,从而增强卵巢癌细胞对顺铂的耐药性。     

LncRNA ITGA9-AS1在卵巢癌中的表达及与顺铂化疗敏感性的关系

目的:观察长链非编码RNA（LncRNA） ITGA9-AS1在卵巢癌中的表达水平,探究其与顺铂化疗敏感性的关系。方法:选取2017年01月至2019年01月本院收治的64例卵巢癌患者,卵巢摘除手术中采集卵巢癌组织,另收集因卵巢囊肿需作手术切除但已证实无瘤细胞的正常卵巢组织标本,采用实时定量PCR（RT-qPCR）检测卵巢癌及正常卵巢组织、顺铂（DDP）耐药卵巢癌细胞SKOV3/DDP、卵巢癌细胞（OVCAR5、OVCAR8、SKOV3）及永生化卵巢上皮细胞（IOSE29、IOSE80）中LncRNA ITGA9-AS1表达水平。采用慢病毒介导LncRNA ITGA9-AS1转染并筛选稳定细胞株,MTT法检测过表达LncRNA ITGA9-AS1对SKOV3、SKOV3/DDP增殖的影响及对不同浓度（0、1、2、4、8、16 mg/L）DDP的敏感性。结果:与正常卵巢组织比较,卵巢癌组织中LncRNA ITGA9-AS1表达水平降低（P＜0.05）。与IOSE29、IOSE80细胞比较,OVCAR5、OVCAR8、SKOV3及SKOV3/DDP细胞中LncRNA ITGA9-AS1表达水平显著降低（P＜0.05）;与SKOV3细胞比较,SKOV3/DDP细胞中LncRNA ITGA9-AS1表达水平显著降低（P＜0.05）。与空白对照组及慢病毒对照组比较,慢病毒过表达组SKOV3、SKOV3/DDP细胞中LncRNA ITGA9-AS1表达水平升高（P＜0.05）,增殖率降低（P＜0.05）,细胞存活率呈DDP浓度依赖降低（P＜0.05）。结论:LncRNA ITGA9-AS1在卵巢癌组织及癌细胞中低表达,过表达LncRNA ITGA9-AS1可增加卵巢癌SKOV3及卵巢癌顺铂耐药细胞SKOV3/DDP的顺铂化疗敏感性。     

As2O3诱导胰腺癌细胞凋亡的相关研究

目的探讨三氧化二砷(arsenictrioxide,As2O3)对胰腺癌细胞的生长的影响及其作用机制。方法采用不同浓度的As2O3处理胰腺癌AsPC-1细胞不同时间(1d、2d、3d),在显微镜和电镜下观察细胞的形态,噻唑蓝(MTT)法和流式细胞分析法测定细胞生长抑制情况;不同浓度的As2O3处理细胞2h、5h、8h、12h、24h后,用DCFH-DA标记细胞后检测荧光强度反映细胞内过氧化氢(H2O2)水平。结果1μmol/LAs2O3处理AsPC-1细胞24h即呈现明显的生长抑制和凋亡特征。As2O3对细胞内H2O2水平影响的结果显示As2O3处理AsPC-1细胞2h后,细胞内H2O2的水平即明显升高(10μmol/L组达79%),到5h达到高峰(增高169%),12h后下降,24h水平与对照组接近,且H2O2水平的变化对As2O3有明显的剂量依赖性。结论As2O3能诱导AsPC-1细胞凋亡,其机制可能与细胞内的H2O2水平改变有关,并且H2O2积累是As2O3诱导细胞凋亡途径中的早期事件,H2O2可能在As2O3诱导肿瘤细胞凋亡途径中扮演类似第二信使的角色。     

拓扑替康诱导卵巢癌COC1/ DDP 细胞凋亡的分子机制初步探讨

  目的 探讨拓扑替康(TPT)对人卵巢癌顺铂耐药细胞株COC1／DDP的杀伤和诱导凋亡活性及其作用机制。方法 MTT比色法与软琼脂克隆形成测定TPT对人卵巢癌顺铂耐药细胞株COC1／DDP的杀伤效应；TUNEL法和流式细胞仪研究TPT对靶细胞的凋亡诱导作用；Western blot检测TPT对COC1／DDP细胞内bcl-2、bax和caspase-3基因表达的影响以及caspase-3活性的改变。结果 TPT对COC1／DDP细胞有明显细胞毒性作用，不仅有剂量依赖性，也存在明显的时间依赖性；COC1／DDP细胞在TPT作用后出现特征性凋亡形态特征，且凋亡率由8．54％上升为23．16％(P<0．05)。TPT不影响COC1／DDP细胞内bcl-2蛋白表达，却明显增加19ax蛋白和caspase-3蛋白表达，并提高caspase-3活性。结论 TPT对人卵巢癌顺铂耐药细胞株COC1／DDP有明显的杀伤和促凋亡作用，其机制可能依赖于细胞内bax蛋白表达增高和caspase-3的活化。     

人参皂甙 Rh2 联合顺铂对食管癌细胞的杀伤作用

目的：体外观察低剂量人参皂甙Rh2（ginsenoside Rh2,GS-Rh2）对抗肿瘤药顺铂（cisplatin,DDP）杀伤人食管癌细胞株Eca109的影响,并探讨其可能的作用机制.方法：人食管癌细胞Eca109经培养符合条件后,分别加用0.3μg/ml DDP（A组）、20μmol/ml GS-Rh2（B组）、0.3μg/ml DDP＋20μmol/ml GS-Rh2（C组）、对照组（D组）处理48小时.流式细胞仪检测各组细胞凋亡率和死亡率.高效液相色谱检测各组细胞内DDP的药物浓度.蛋白质印迹法检测各组细胞中p53表达.结果：细胞凋亡率和死亡率C组显著高于A、B两组（P＜0.05）;C组细胞内DDP的药物浓度显著高于A组细胞（P＜0.05）;C组细胞p53表达水平低于A组（P＜0.05）.结论：体外低剂量GS-Rh2能增强DDP对人食管癌细胞Eca109的杀伤作用,其作用机制可能与降低细胞内p53表达有关.     

CIK细胞联合顺铂对卵巢癌耐药细胞SKOV3/CDDP的杀伤作用

目的：探讨细胞因子诱导的杀伤（cytokine-induced killer,CIK）细胞联合顺铂对卵巢癌顺铂耐药细胞SKOV3/CD-DP小鼠腹腔移植模型的体内外杀伤作用。方法：取正常人的外周血单个核细胞（PBMC）,加入IFN-γ、IL-2、CD3McAb和IL-1,体外诱导成CIK细胞。用MTT法测定CIK细胞、顺铂及两者联合对人卵巢癌细胞株SKOV3/CDDP细胞的杀伤活性。在SCID小鼠腹腔内接种卵巢癌SKOV3/CDDP细胞,观察CIK细胞对荷瘤鼠的体内抑瘤作用,并用RIA法检测各组血清中CA125的含量。结果：顺铂对SKOV3/CDDP细胞的IC50为315.5μg/ml,较其对SKOV3细胞的IC50（85μg/ml）增加了4倍;CIK细胞对SKOV3与SKOV3/CDDP细胞的杀伤活性均在48h最高,且随效靶比的增高而增强,其对SKOV3/CDDP细胞的杀伤活性明显高于SKOV3（P〈0.05）。仅25μg/ml的顺铂即可使效靶比为12.5∶1组中的联合杀伤率比单纯顺铂组增加5.8倍,比单纯加CIK（相同效靶比）细胞杀伤率增加2.3倍,且随效靶细胞比值和顺铂浓度的升高呈依赖性增加。与对照组相比,CIK组与CIK＋顺铂组均能显著抑制癌性结节的生长,抑瘤率达100%,且3组SCID鼠血中CA125值有显著性差异（P〈0.05）。结论：正常人CIK细胞联合顺铂对清除晚期卵巢癌表达耐药蛋白的腹腔种植转移病灶可能会有较好的效果,并有希望成为前景良好的综合治疗方案。     

Enhanced cisplatin cytotoxicity by disturbing the nucleotide excision repair pathway in ovarian cancer cell lines

Ovarian cancer is the leading cause of death among women from gynecological malignancies inthe United States. Resistance to the chemotherapeutic agent cisplatin isa major limitation for the successful treatment of ovarian cancer. In an effort to overcome the cisplatin resistance problem in ovarian cancer treatment, we have sought to enhance cisplatin cytotoxicity by perturbing the nucleotide excision repair (NER) pathway. The NER pathway is responsible for repairing cisplatin bound to DNA. Expression of one of the NER components, ERCC1, is correlated with cisplatin drug resistance. Hence, we targeted ERCC1 by antisense RNA methodologies, and we show that we could sensitize a relatively sensitive A2780 cell line and also the highly resistant OVCAR10 cell line to cisplatin by expressing antisense ERCC1 RNA in them as measured with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. The A2780 cell lines expressing antisense ERCC1 had 1.9-8.1-fold enhancements in cisplatin sensitivity. The OVCAR10 antisense ERCC1 cell lines had IC(50) values ranging from 2.28 microM to 2.7 microM cisplatin as compared with 9.52 micro M for control OVCAR10 cells. The OVCAR10 antisense ERCC1 cells also show reduced DNA-damage repair capacity as assessed by host cell reactivation. Furthermore, immunocompromised mice transplanted with the antisense cell lines survived longer than the mice bearing control cells after response to cisplatin treatment. These data suggest that it is possible to substantially enhance the cisplatin cytotoxicity by disturbing the NER pathway in cisplatin-resistant cell lines and to enhance the survival capacity of mice in an ovarian cancer xenograft model.     

Arsenic compounds induce cytotoxicity and apoptosis in cisplatin-sensitive and -resistant gynecological cancer cell lines

PURPOSE: Arsenic compounds have been found to be effective in the treatment of acute promyelocytic leukemia through the downregulation of bcl-2 expression. Resistant ovarian cancer cells often overexpress bcl-2 or p53 proteins or both. We hypothesized that arsenic compounds, such as As2O3 and As2S3, could also be active against gynecological cancers resistant to conventional chemotherapy. METHODS: We investigated the effects of these two arsenic compounds in vitro on ovarian cancer cell lines sensitive (OVCAR, GG, JAM) and resistant (CI80-13S) to cisplatin (CDDP) and on human cervical cancer cell lines (HeLa) in comparison with their effects on human fibroblasts (HF). A fluorometric assay based on measurements of fluorescein diacetate (FDA) in cells was used to determine cell viability. Apoptosis was assessed in terms of cell morphology, by flow cytometry and by a DNA fragmentation assay. RESULTS: Treatment of each cell line with the As2O3 or As2S3 led to a marked dose-dependent decrease in cell growth. The IC50 of the two compounds indicated a significantly greater cytotoxic effect against all the cancer cells tested than against the normal HF. At a clinically achievable concentration (2 microM), As2O3 selectively inhibited the growth and induced apoptosis in CI80-13S, OVCAR and HeLa cells but had no significant apoptotic effect on GG or JAM cells or HF. Following treatment with 5 microM As2S3, the CI80-13S, OVCAR and HeLa cells also exhibited growth inhibition and induction of apoptosis. CONCLUSIONS: Arsenic compounds (As2O3 and As2S3) can inhibit growth and induce apoptosis in human ovarian and cervical cancer cells at clinically achievable concentrations, indicating that As2O3 and As2S3 could be effective in the treatment of gynecological cancer.     

Characterizations of irofulven cytotoxicity in combination with cisplatin and oxaliplatin in human colon, breast, and ovarian cancer cells

Purpose: This study assessed the cytotoxic effects of irofulven in combination with oxaliplatin and cisplatin in a panel of human cancer cell lines. Methods: Growth inhibition studies were performed using the human HT29 colon cancer cell line, irofulven-resistant derivative HT29/IF2, breast cancer cell line MCF7, and ovarian cancer line CAOV3. Irofulven–oxaliplatin combinations were compared with irofulven–cisplatin combinations in the same cell lines using similar experimental settings. Cells were exposed for 1 h to irofulven and then for 24 h to oxaliplatin or cisplatin and vice versa. Results: Single agent irofulven displayed cytotoxic effects against human colon HT29 cells, human breast cancer cell lines including MCF7, SKBR3, and ZR-75-1, and human ovarian cancer cell lines CAOV3, OVCAR3, and IGROV1, with OVCAR3 being the most sensitive cancer cell line (IC₅₀: 2.4 μM). In all tested cell lines the oxaliplatin–irofulven combination led to clear evidence of synergistic activity. In HT29 and HT29/IF2, the sequence oxaliplatin followed by irofulven appears to be the most effective whereas in MCF7 cells, irofulven given prior to or simultaneously with oxaliplatin is more effective than the other schedule. The combination displays additive activity toward CAOV3 ovarian cells when irofulven was administered prior to or simultaneously with oxaliplatin and partially synergistic when oxaliplatin was followed by irofulven. In most of the cell lines, the sequence oxaliplatin followed by irofulven appears to be the most effective as compared to other schedules. A combination of irofulven with cisplatin has the same efficacy as with oxaliplatin for the same cell lines. Cell cycle studies show that irofulven increases the proportion of cells in the S phase. Cisplatin–irofulven and oxaliplatin–irofulven combinations block cells in G1/S and potently induce apoptosis. Conclusion: Irofulven displays synergistic antiproliferative and pro-apoptotic effects when combined with oxaliplatin over a broad range of concentrations in human colon and breast cancer cells. Acquired resistance to irofulven has limited impact on the effects of cisplatin–irofulven and oxaliplatin–irofulven combinations. Based on these data, irofulven–oxaliplatin and cisplatin–irofulven combinations will be further explored in clinical trials, favoring the use schedules of oxaliplatin given prior to irofulven in patients with cancer.     

siRNA沉默结肠癌转移相关基因1表达对人类卵巢癌细胞株侵袭转移的影响

目的 检测结肠癌转移相关基因1（MACC1）在卵巢癌细胞株中的表达,并探讨用siRNA技术抑制其表达对卵巢癌细胞生物学行为的影响.方法 应用实时荧光定量PCR （RT-qPCR）及Western blot法检测OVCAR3、ES-2、SKOV3、HO-8910卵巢癌细胞株中MACC1的表达,合成MACC1特异性siRNA并转染OVCAR3细胞,利用RT-qPCR筛选并鉴定MACC 1基因有效沉默后,应用体外黏附实验、Transwell迁移及侵袭实验、体外血管拟态实验检测MACC1基因沉默后OVCAR3细胞的体外黏附、迁移、侵袭及血管生成能力的变化.结果 OVCAR3细胞较其他卵巢癌细胞株高表达MACC1.MACC1基因沉默后,OVCAR3细胞的体外黏附能力受到不同程度的抑制;Transwell迁移实验中,MACC1 siRNA干扰的OVCAR3细胞（干扰组）转入底层膜的细胞数为（245.5±12.8）个,低于阴性对照组和空白对照组[分别为（500.3±16.5）、（496.3±13.1）个,均P＜0.05]; Transwell侵袭实验中,干扰组转入底层膜的细胞数为（185.3±14.1）个,低于阴性对照组和空白对照组[分别为（405.7±9.1）、（416.3±11.5）个,均P＜0.05];体外血管拟态显示干扰组细胞多呈散在分布,连接减少,形成的完整结构少.结论 利用siRNA技术抑制MACC1基因表达可有效抑制卵巢癌OVCAR3细胞的体外转移和侵袭能力,MACC1有望成为卵巢癌治疗的靶基因.     

RAD001 inhibits human ovarian cancer cell proliferation, enhances cisplatin-induced apoptosis, and prolongs survival in an ovarian cancer model.

PURPOSE: mTOR (mammalian target of rapamycin) plays a central role in regulating cell growth and cell cycle progression and is regarded as a promising therapeutic target. We examined whether mTOR inhibition by RAD001 (everolimus) is therapeutically efficacious in the treatment of ovarian cancer as a single agent and in combination with cisplatin. EXPERIMENTAL DESIGN: Using four human ovarian cancer cell lines, we determined the effect of RAD001 by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Western blot, and apoptosis assays. We evaluated the association between phospho-AKT/mTOR activity and RAD001 sensitivity. We also determined the effect of RAD001 on tumor growth and malignancy using mice inoculated with human ovarian cancer cells. RESULTS: RAD001 markedly inhibited cell proliferation of human ovarian carcinoma cells with high AKT activity (OVCAR10 and SKOV-3), but the effect was minimal in cells with low AKT activity (OVCAR4 and OVCAR5). Sensitivity to RAD001 was independent of p53 expression. RAD001 inhibited the phosphorylation of downstream 4E-BP1 and p70S6 kinase and attenuated the expression of Myc. RAD001 also attenuated the expression of HIF-1 alpha and vascular endothelial growth factor, important factors in angiogenesis and tumor invasiveness. RAD001 enhanced cisplatin-induced apoptosis in cells with high AKT/mTOR activity, with minimal effect in cells with low AKT-mTOR activity. Mouse xenografts of SKOV-3 cells revealed that RAD001 inhibits tumor growth, angiogenesis, and i.p. dissemination and ascites production and prolongs survival. Moreover, treatment with RAD001 significantly enhanced the therapeutic efficacy of cisplatin in vivo. CONCLUSION: These results indicate that RAD001 could have therapeutic efficacy in human ovarian cancers with hyperactivated AKT/mTOR signaling.