Reovirus infection of cancer cells is not due to activated Ras pathway.
Reovirus infection of cancer cells is not due to activated Ras pathway.
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呼肠孤病毒感染癌细胞并非由于激活Ras途径所致。
DOI：
10.1038/cgt.2008.84
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发表时间：
2009
影响因子：
6.4
通讯作者：
Holland,JF
中科院分区：
文献类型：
--
作者：
Song,L;Ohnuma,T;Gelman,IH;Holland,JF
Lee和他的同事1，2声称呼肠孤病毒选择性地杀死具有激活的Ras途径的细胞：激活的Ras或Ras途径的激活元件抑制双链RNA激活蛋白激酶（PKR）激活，从而允许病毒蛋白质合成。呼肠孤病毒作为一种潜在的抗癌剂仍在积极研究中。我们在八种人类肿瘤细胞系中检测了呼肠孤病毒诱导的细胞病变效应（CPE）。未转化的NIH/3 T3和LLC-MK2细胞系作为对照。NIH/3 T3是小鼠成纤维细胞系; 3 LLC-MK2细胞系来源于成年恒河猴的肾脏。4呼肠孤病毒血清型3可以感染LLCMK 2、许多人肿瘤细胞系和几种未转化的鼠细胞系如L929，5，但不感染NIH/3 T3。我们试图将CPE和通过聚丙烯酰胺凝胶电泳测量的细胞内病毒RNA与通过蛋白质印迹法测量的单个细胞系中的相对Ras蛋白水平相关联。此外，尝试通过免疫印迹法检测总的和磷酸化的双链RNA激活蛋白激酶和磷酸化的真核起始因子-2 α（eIF-2α），将病毒诱导的翻译控制途径与CPE联系起来。6我们证实在8个肿瘤细胞系和LLC-MK 2细胞中观察到的CPE是由于病毒感染所致。NIH/3 T3细胞未表现出可检测到的病毒RNA，这与缺乏感染性相关。病毒感染前后Ras表达水平变化不大。Ras蛋白水平与CPE程度相关（NIH/3 T3细胞除外）。Ras在DND-1A黑色素瘤、2780卵巢癌和NIH/3 T3细胞系中的表达水平相同。总双链RNA激活蛋白激酶或磷酸化双链RNA激活蛋白激酶的表达与CPE和Ras表达均不相关。磷酸化真核起始因子-2 α水平升高仅在4种细胞系（DND-1A、U87 MG胶质母细胞瘤、NIH/3 T3和LLC-MK 2）中与CPE呈负相关。在其余6个细胞系（2780、Hep-2喉鳞状细胞癌、MCF-7乳腺癌、人绒毛膜癌、U87 MG. wtEGFR
Lee and his co-workers 1, 2 claimed that reovirus selectively killed cells with an activated Ras pathway: activated Ras or an activated element of the Ras pathway inhibited double-stranded RNA-activated protein kinase (PKR) activation, thereby allowing viral protein synthesis. Reovirus remains actively pursued as a potential anticancer agent. We have examined the reovirus-induced cytopathic effect (CPE) in eight human tumor cell lines. Untransformed NIH/3T3 and LLC-MK2 cell lines served as controls. NIH/3T3 is a mouse fibroblast cell line; 3 the LLC-MK2 cell line was derived from kidneys of adult Rhesus monkeys. 4 Reovirus serotype 3 can infect LLCMK2, many human tumor cell lines and several untransformed murine cell lines such as L929, 5 but not NIH/3T3. We sought to correlate CPE, and intracellular viral RNA measured by polyacrylamide gel electrophoresis, with relative Ras protein levels in the individual cell lines, measured by western blotting. In addition, attempts were made to correlate virus-induced translational control pathways with CPE by immunoblotting for total and phosphorylated double-stranded RNA-activated protein kinase and phosphorylated eukaryotic initiation factor-2α (eIF-2α). 6We confirmed that the CPE observed in eight tumor cell lines and LLC-MK2 cells was due to the viral infection. NIH/3T3 cells exhibited no detectable viral RNA, correlating with a lack of infectivity. Ras expression levels showed little change before and after the virus infection. Ras protein levels correlated with the degree of CPE (except in NIH/3T3 cells). The level of Ras expression was the same in DND-1A melanoma, 2780 ovarian carcinoma and NIH/3T3 cell lines. The expression of total double-stranded RNA-activated protein kinase or phosphorylated double-stranded RNA-activated protein kinase correlated with neither CPE nor Ras expression. Elevated levels of phosphorylated eukaryotic initiation factor-2α correlated inversely with CPE only in four cell lines (DND-1A, U87MG glioblastoma, NIH/3T3 and LLC-MK2). In the remaining six cell lines (2780, Hep-2 laryngeal squamous cell carcinoma, MCF-7 breast cancer, JAR choriocarcinoma, U87MG. wtEGFR