Myxoma virus-mediated oncolysis of ascites-derived human ovarian cancer cells and spheroids is impacted by differential AKT activity.
Myxoma virus-mediated oncolysis of ascites-derived human ovarian cancer cells and spheroids is impacted by differential AKT activity.
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粘液瘤病毒介导的腹水来源的卵巢癌细胞和球体的溶瘤作用受到差异 AKT 活性的影响。
DOI：
10.1016/j.ygyno.2012.01.048
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发表时间：
2012-05
影响因子：
4.7
通讯作者：
Shepherd TG
中科院分区：
文献类型：
--
作者：
Correa RJ;Komar M;Tong JG;Sivapragasam M;Rahman MM;McFadden G;Dimattia GE;Shepherd TG
我们认为转移性上皮性卵巢癌(EOC)是溶瘤剂粘液瘤病毒(MYXV)的潜在治疗靶点。从患者腹水中分离出原代EOC细胞，并将其培养为贴壁细胞或使用超低贴壁培养皿悬浮培养。用表达绿色荧光蛋白的MYXV感染细胞和球体。通过荧光显微镜、病毒滴定和免疫印迹检测M-T7和M130病毒蛋白的表达，并用AlamarBlue法检测细胞活力。用Akti-1/2(5μM)和雷帕霉素(20 NM)检测PI3K-akt信号在介导MYXV感染中的作用。在贴壁培养中生长的腹水来源的EOC细胞可被MYXV感染有效地杀死。悬浮培养的EoC细胞形成三维EoC球体，很容易使MYXV进入细胞，但不受晚期MYXV感染的细胞病变影响。在重新附着时(以模拟继发性转移)，EoC球体对MYXV介导的肿瘤分解重新敏感。促进MYXV有效感染的关键决定因素是激活的PI3K-AKT信号通路的存在。用AKT特异性抑制剂Akti-1/2治疗可减少单层EoC细胞和球体的感染。直接感染新鲜采集的腹水表明，54.5%的患者样本对MYXV介导的溶瘤细胞杀伤敏感。我们还证明，腹水中存在的S因子可能对MYXV感染和EOC细胞溶瘤产生负面影响，这可能是由于内源性AKT活性下调所致。AKT的不同活性是在转移计划的关键步骤中调节MYXV介导的EOC球体溶瘤的机制基础。此外，我们提供了第一个证据，证明MYXV溶瘤疗法可能对相当大比例的卵巢癌转移患者有效。
We propose that metastatic epithelial ovarian cancer (EOC) is a potential therapeutic target for the oncolytic agent, Myxoma virus (MYXV). Primary EOC cells were isolated from patient ascites and cultured as adherent cells or in suspension using Ultra Low-Attachment dishes. MYXV expressing green fluorescent protein was used to infect cells and spheroids. Infection was monitored by fluorescence microscopy, viral titering and immunoblotting for M-T7 and M130 virus protein expression, and cell viability by alamarBlue assay. Akti-1/2 (5 μM) and rapamycin (20 nM) were used to assay the role of PI3K–AKT signaling in mediating MYXV infection. Ascites-derived EOC cells grown in adherent culture are effectively killed by MYXV infection. EOC cells grown in suspension to form three-dimensional EOC spheroids readily permit MYXV entry into cells, yet are protected from the cytopathic effects of late MYXV infection. Upon reattachment (to model secondary metastasis), EOC spheroids are re sensitized to MYXV-mediated oncolysis. The critical determinant that facilitates efficient MYXV infection is the presence of an activated PI3K-AKT signaling pathway. Treatment with the specific AKT inhibitor Akti-1/2 reduces infection of monolayer EOC cells and spheroids. Direct infection of freshly collected ascites demonstrated that 54.5% of patient samples were sensitive to MYXV-mediated oncolytic cell killing. We also demonstrate that factor(s) present in ascites may negatively impact MYXV infection and oncolysis of EOC cells, which may be due to a down-regulation in endogenous AKT activity. Differential activity of AKT serves as the mechanistic basis for regulating MYXV-mediated oncolysis of EOC spheroids during key steps of the metastatic program. In addition, we provide the first evidence that MYXV oncolytic therapy may be efficacious for a significant proportion of ovarian cancer patients with metastatic disease.
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DOI：
10.1038/nrmicro1099
发表时间：
2005-03
期刊：
Nature reviews. Microbiology
影响因子：
--
作者：
通讯作者：
--
影响因子：
7
作者：
Kuk, Cynthia;Kulasingam, Vathany;Diamandis, Eleftherios P.
通讯作者：
Diamandis, Eleftherios P.
影响因子：
4.7
作者：
Correa, Rohann J. M.;Peart, Teresa;Shepherd, Trevor G.
通讯作者：
Shepherd, Trevor G.
影响因子：
11.2
作者：
Galanis E;Hartmann LC;Cliby WA;Long HJ;Peethambaram PP;Barrette BA;Kaur JS;Haluska PJ Jr;Aderca I;Zollman PJ;Sloan JA;Keeney G;Atherton PJ;Podratz KC;Dowdy SC;Stanhope CR;Wilson TO;Federspiel MJ;Peng KW;Russell SJ
通讯作者：
Russell SJ
影响因子：
12.4
作者：
Jiang, H;Wang, Z;Amalfitano, A
通讯作者：
Amalfitano, A