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[iTRAQ] 卵巢癌-biomarker研究-iTRAQ技术

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发表于 2015-12-17 17:51:47 | 显示全部楼层 |阅读模式

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Mol Cell Proteomics. 2013 Feb;12(2):356-68. doi: 10.1074/mcp.M112.019521. Epub 2012 Nov 19.
Proteomic analysis of temporally stimulated ovarian cancer cells for biomarker discovery.

IF=
14年6.564
13年7.254
12年7.251
11年7.398

研究关键词卵巢癌,生物标记物, iTRAQ,lysosomal-associated membrane protein 1 (LAMP-1),EGFR,PI3K/Akt

研究概要
本研究目的在于通过蛋白质组学技术探索卵巢癌对于酪氨酸激酶抑制剂或EGFR单克隆抗体治疗敏感性的生物标记物。 研究对象为人卵巢癌细胞系OVCAR3,分别用EGF、LY294002(PI3K/Akt抑制剂)、EGF+LY294002处理细胞,应用MTT检测各组细胞的增值活性,并分别于处理12h、24h、48h时留取收集细胞及基质,用iTRAQ技术检测每个时间点时4组细胞(以上3个处理组及对照组)及4组基质的蛋白表达情况,即进行3个8标的iTRAQ检测,通过生物信息学分析发现,LAMP1可能可以作为卵巢癌的生物标记物,进一步扩大样本量,利用组织芯片及Western blot技术验证该结论。该研究主要结论为发现LAMP1可能可以用来预估酪氨酸激酶抑制剂或EGFR单克隆抗体对卵巢癌治疗的有效性。

Abstract
While ovarian cancer remains the most lethal gynecological malignancy in the United States, there are no biomarkers available that are able to predict therapeutic responses to ovarian malignancies. One major hurdle in the identification of useful biomarkers has been the ability to obtain enough ovarian cancer cells from primary tissues diagnosed in the early stages of serous carcinomas, the most deadly subtype of ovarian tumor. In order to detect ovarian cancer in a state of  hyperproliferation, we analyzed the implications of molecular signaling cascades in the ovarian cancer cell line OVCAR3 in a temporal manner, using a mass-spectrometry-based proteomics approach. OVCAR3 cells were treated with EGF(1), and the time course of cell progression was monitored based on Akt phosphorylation and growth dynamics. EGF-stimulated Akt phosphorylation was detected at 12 h post-treatment, but an effect on proliferation was not observed until 48 h post-exposure. Growth-stimulated cellular lysates were analyzed for protein profiles between treatment groups and across time points using iTRAQ labeling and mass spectrometry. The protein response to EGF treatment was identified via iTRAQ analysis in EGF-stimulated lysates relative to vehicle-treated specimens across the treatment time course. Validation studies were performed on one of the differentially regulated proteins, lysosomal-associated membrane protein 1 (LAMP-1), in human tissue lysates and ovarian tumor tissue sections. Further, tissue microarray analysis was performed to demarcate LAMP-1 expression across different stages of epithelial ovarian cancers. These data support the use of this approach for the efficient identification of tissue-based markers in tumor development related to specific signaling pathways. LAMP-1 is a promising biomarker for studies of the progression of EGF-stimulated ovariancancers and might be useful in predicting treatment responses involving tyrosine kinase inhibitors or EGF receptor monoclonal antibodies.


PMID: 23172893  PMCID: PMC3567859

全文链接
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3567859/







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发表于 2015-12-17 18:30:30 微信发帖 | 显示全部楼层
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