Reovirus exerts potent oncolytic effects in head and neck cancer cell lines that are independent of signalling in the EGFR pathway.
Reovirus exerts potent oncolytic effects in head and neck cancer cell lines that are independent of signalling in the EGFR pathway.
复制标题
异病毒在头颈癌细胞系中发挥有效的溶瘤作用，这与EGFR途径中的信号无关。
DOI：
10.1186/1471-2407-12-368
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发表时间：
2012-08-24
期刊：
影响因子：
3.8
通讯作者：
Harrington KJ
中科院分区：
文献类型：
--
作者：
Twigger K;Roulstone V;Kyula J;Karapanagiotou EM;Syrigos KN;Morgan R;White C;Bhide S;Nuovo G;Coffey M;Thompson B;Jebar A;Errington F;Melcher AA;Vile RG;Pandha HS;Harrington KJ
呼肠孤病毒利用Ras下游的异常信号传导介导肿瘤特异性溶瘤。由于~90%的头颈部鳞状细胞癌（SCCHN）过表达EGFR，SCCHN细胞系对溶瘤呼肠孤病毒敏感，我们对15种头颈部癌细胞系中呼肠孤病毒的作用进行了详细分析。研究了入组前和入组后事件，试图定义预测呼肠孤病毒敏感性/耐药性的生物标志物。特别是，我们分析了EGFR/Ras信号在确定SCCHN中病毒介导的细胞毒性中的作用。为了测试EGFR途径活性是否预测对呼肠孤病毒的敏感性增加，进行了通过MTT测定的呼肠孤病毒IC 50与通过蛋白质印迹和FACS测定的EGFR水平之间的相关性分析。通过MTT测定分析EGFR信号传导的抑制或刺激对呼肠孤病毒溶瘤作用的影响，并通过TCID 50测定分析病毒生长。我们接下来分析了通过p38 MAPK、PI 3-K或MEK的特异性抑制剂抑制Ras下游信号传导对通过MTT测定检测的呼肠孤病毒杀伤的影响。PKR在呼肠孤病毒杀伤中的作用也通过使用2-氨基嘌呤阻断PKR和通过MTT测定测定细胞存活来确定。通过蛋白质印迹法分析caspase 3切割来检测SCCHN对呼肠孤病毒的凋亡反应。呼肠孤病毒敏感性和EGFR水平之间的相关性分析显示没有关联。中间亚病毒和核心颗粒显示出与完整呼肠孤病毒相同的感染性/细胞毒性。因此，未通过细胞进入确定灵敏度。在4个细胞系中，肿瘤溶解和病毒生长均不受EGFR信号传导抑制或刺激的影响。Ras下游信号传导的抑制没有消除呼肠孤病毒的溶瘤作用，此外，使用2-氨基嘌呤调节PKR没有改变抗性细胞系中呼肠孤病毒的敏感性。在感染的细胞中未检测到半胱天冬酶3裂解，在泛半胱天冬酶抑制的细胞中观察到溶瘤作用。总之，呼肠孤病毒在广泛的SCCHN细胞系中具有强效溶瘤作用。通过分析EGFR/Ras/MAPK通路来定义敏感性/耐药性的尝试未能提供明确的反应预测生物标志物。目前正在对体外和临床研究的材料进行进一步分析，以进一步阐明这一问题。
Reovirus exploits aberrant signalling downstream of Ras to mediate tumor-specific oncolysis. Since ~90% squamous cell carcinomas of the head and neck (SCCHN) over-express EGFR and SCCHN cell lines are sensitive to oncolytic reovirus, we conducted a detailed analysis of the effects of reovirus in 15 head and neck cancer cell lines. Both pre- and post-entry events were studied in an attempt to define biomarkers predictive of sensitivity/resistance to reovirus. In particular, we analysed the role of EGFR/Ras signalling in determining virus-mediated cytotoxicity in SCCHN. To test whether EGFR pathway activity was predictive of increased sensitivity to reovirus, correlative analyses between reoviral IC50 by MTT assay and EGFR levels by western blot and FACS were conducted. Inhibition or stimulation of EGFR signalling were analysed for their effect on reoviral oncolysis by MTT assay, and viral growth by TCID50 assay. We next analysed the effects of inhibiting signalling downstream of Ras, by specific inhibitors of p38MAPK, PI3-K or MEK, on reoviral killing examined by MTT assay. The role of PKR in reoviral killing was also determined by blockade of PKR using 2-aminopurine and assaying for cell survival by MTT assay. The apoptotic response of SCCHN to reovirus was examined by western blot analysis of caspase 3 cleavage. Correlative analyses between reoviral sensitivity and EGFR levels revealed no association. Intermediate sub-viral and core particles showed the same infectivity/cytotoxicity as intact reovirus. Therefore, sensitivity was not determined by cell entry. In 4 cell lines, oncolysis and viral growth were both unaffected by inhibition or stimulation of EGFR signalling. Inhibition of signalling downstream of Ras did not abrogate reoviral oncolysis and, in addition, modulation of PKR using 2-aminopurine did not alter reovirus sensitivity in resistant cell lines. Caspase 3 cleavage was not detected in infected cells and oncolysis was observed in pan-caspase inhibited cells. In summary, reovirus is potently oncolytic in a broad panel of SCCHN cell lines. Attempts to define sensitivity/resistance by analysis of the EGFR/Ras/MAPK pathway have failed to provide a clear predictive biomarker of response. Further analysis of material from in vitro and clinical studies is ongoing in an attempt to shed further light on this issue.
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影响因子：
2.7
作者：
HASHIRO, G;LOH, PC;YAU, JT
通讯作者：
YAU, JT
影响因子：
5.1
作者：
Errington, F.;White, C. L.;Melcher, A. A.
通讯作者：
Melcher, A. A.
影响因子：
4.8
作者：
Favata, MF;Horiuchi, KY;Trzaskos, JM
通讯作者：
Trzaskos, JM
影响因子：
6.1
作者：
Cassell A;Grandis JR
通讯作者：
Grandis JR
DOI：
10.1158/1078-0432.ccr-10-0054
发表时间：
2010-06-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Harrington KJ;Karapanagiotou EM;Roulstone V;Twigger KR;White CL;Vidal L;Beirne D;Prestwich R;Newbold K;Ahmed M;Thway K;Nutting CM;Coffey M;Harris D;Vile RG;Pandha HS;Debono JS;Melcher AA
通讯作者：
Melcher AA