Measuring soluble forms of extracellular cytokeratin 18 identifies both apoptotic and necrotic mechanisms of cell death produced by adenoviral-mediated interferon alpha: possible use as a surrogate marker.
Measuring soluble forms of extracellular cytokeratin 18 identifies both apoptotic and necrotic mechanisms of cell death produced by adenoviral-mediated interferon alpha: possible use as a surrogate marker.
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DOI：
10.1038/cgt.2009.2
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发表时间：
2009-07
影响因子：
6.4
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中科院分区：
文献类型：
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腺病毒介导人干扰素α-2b（Ad-IFN）介导的人膀胱癌细胞通过直接和间接机制产生肿瘤特异性细胞死亡。间接机制涉及所产生的分泌型IFN，其杀死IFN蛋白敏感的癌细胞以及对邻近癌细胞具有细胞毒性的尚未鉴定的旁观者因子。Ad-IFN在癌细胞中的转染和表达导致直接细胞杀伤。由于细胞角蛋白18（CK 18）的分子形式，无论半胱天冬酶裂解与否，都分别与细胞凋亡或坏死性细胞死亡相关，因此我们确定了IFNα蛋白或Ad-IFN治疗膀胱癌细胞后是否可以观察到CK 18的一种或两种形式的增加。M30和M65 ELISA定量（分别用于CK 18相关凋亡和坏死细胞死亡的测定）用作产生的细胞死亡的替代标志物。在IFN蛋白敏感的RT 4膀胱癌细胞中，IFN主要产生M30相关的细胞死亡，而Ad-IFN治疗导致高水平的M30和M65。相比之下，Ad-IFN处理的细胞的条件培养基，无论是来自正常人尿路上皮细胞或膀胱癌细胞，当添加到IFN蛋白抗性KU 7或UC 9膀胱癌细胞中时，主要引起M30水平的增加，这表明存在于CM中的旁观者因子主要产生凋亡性细胞死亡。此外，当用Ad-IFN处理IFN蛋白抗性KU 7和UC 9细胞时，观察到M65水平显著增加，高于对M30观察到的水平，再次表明Ad-IFN也产生了额外的坏死相关细胞死亡。Ad-IFN处理后，正常尿路上皮细胞没有表现出细胞毒性，M30或M65也没有增加。由于膀胱内Ad-IFN治疗目前正在评估其在浅表性膀胱癌中的疗效，因此在Ad-IFN治疗之前和之后的各个时间点测量尿液中的M30和M65水平不仅可以提供疗效的生物标志物，而且还可以提供个体患者肿瘤中由各种细胞杀伤机制产生的不同类型和比例的细胞杀伤的证据。
Adenoviral transduction of human bladder cancer cells with human interferon α-2b (Ad-IFN) produces cancer specific cell death via direct and indirect mechanisms. The indirect mechanisms involve the secreted IFN produced which kill IFN protein sensitive cancer cells as well as yet unidentified bystander factors which are cytotoxic to neighboring cancer cells. The direct cell kill results from transfection and expression of Ad-IFN in the cancer cells. Since the molecular forms of cytokeratin 18 (CK18), either caspase cleaved or not, have been associated with apoptotic or necrotic cell death, respectively, we determined if increases in either or both CK18 forms could be observed following IFNα protein or Ad-IFN treatment of bladder carcinoma cells. Quantification of M30 and M65 ELISAs (assays for CK18 associated apoptotic and necrotic cell death, respectively) were used as surrogate markers of the cell death produced. In the IFN protein sensitive RT4 bladder cancer cells IFN produced primarily M30 related cell death whereas Ad-IFN treatment resulted in high levels of both M30 and M65. In contrast, conditioned medium from Ad-IFN treated cells whether from normal human urothelial cells or bladder cancer cells caused mainly increases in M30 levels when added to IFN protein resistant KU7 or UC9 bladder cancer cells, suggesting that the bystander factors present in the CM produced primarily apoptotic cell death. In addition, a significant increase in M65 levels above that observed for M30 was seen when the IFN protein resistant KU7 and UC9 cells were treated with Ad-IFN, again indicating there is additional necrotic related cell death produced by Ad-IFN as well. Normal urothelial cells showed no cytotoxicity nor increases in M30 or M65 following Ad-IFN treatment. Since intravesical Ad-IFN treatment is presently being evaluated for its efficacy in superficial bladder cancer that measurement of M30 and M65 levels in the urine at various time-points before and after Ad-IFN treatment may provide not only a biomarker of efficacy but also evidence for the different types and proportion of cell kill produced by the various mechanisms of cell kill in the tumors of individual patients.
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影响因子：
6.4
作者：
Zhang, X.;Dong, L.;Benedict, W. F.
通讯作者：
Benedict, W. F.
影响因子：
3
作者：
Cummings, J.;Ranson, M.;Dive, C.
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Dive, C.
影响因子：
11.2
作者：
Papageorgiou, A;Lashinger, L;McConkey, DJ
通讯作者：
McConkey, DJ
影响因子：
8
作者：
Chapman, E. J.;Hurst, C. D.;Knowles, M. A.
通讯作者：
Knowles, M. A.
影响因子：
12.4
作者：
Benedict, WF;Tao, ZM;Connor, RJ
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