Fan Alice C, Deb-Basu Debabrita, Orban Mathias W, Gotlib Jason R, Natkunam Yasodha, O'Neill Roger, Padua Rose-Ann, Xu Liwen, Taketa Daryl, Shirer Amy E, Beer Shelly, Yee Ada X, Voehringer David W, Felsher Dean W
Stanford University, Departments of Medicine and Pathology, California, USA.
Nat Med. 2009 May;15(5):566-71. doi: 10.1038/nm.1903. Epub 2009 Apr 12.
Current methods of protein detection are insensitive to detecting subtle changes in oncoprotein activation that underlie key cancer signaling processes. The requirement for large numbers of cells precludes serial tumor sampling for assessing a response to therapeutics. Therefore, we have developed a nanofluidic proteomic immunoassay (NIA) to quantify total and low-abundance protein isoforms in nanoliter volumes. Our method can quantify amounts of MYC oncoprotein and B cell lymphoma protein-2 (BCL2) in Burkitt's and follicular lymphoma; identify changes in activation of extracellular signal-related kinases-1 (ERK1) and ERK2, mitogen-activated kinase-1 (MEK), signal transducer and activator of transcription protein-3 (STAT3) and STAT5, c-Jun N-terminal kinase (JNK) and caspase-3 in imatinib-treated chronic myelogeneous leukemia (CML) cells; measure an unanticipated change in the phosphorylation of an ERK2 isomer in individuals with CML who responded to imatinib; and detect a decrease in STAT3 and STAT5 phosphorylation in individuals with lymphoma who were treated with atorvastatin. Therefore, we have described a new and highly sensitive method for determining oncoprotein expression and phosphorylation in clinical specimens for the development of new therapeutics for cancer.
当前的蛋白质检测方法对于检测作为关键癌症信号传导过程基础的癌蛋白激活的细微变化并不敏感。由于需要大量细胞，因此无法进行连续肿瘤采样以评估对治疗的反应。因此，我们开发了一种纳米流体蛋白质组免疫测定法（NIA），用于在纳升体积中定量总蛋白和低丰度蛋白异构体。我们的方法可以定量伯基特淋巴瘤和滤泡性淋巴瘤中MYC癌蛋白和B细胞淋巴瘤蛋白-2（BCL2）的含量；鉴定伊马替尼治疗的慢性粒细胞白血病（CML）细胞中细胞外信号相关激酶-1（ERK1）和ERK2、丝裂原活化激酶-1（MEK）、信号转导和转录激活蛋白-3（STAT3）和STAT5、c-Jun N端激酶（JNK）和半胱天冬酶-3激活的变化；测量对伊马替尼有反应的CML患者中ERK2异构体磷酸化的意外变化；并检测用阿托伐他汀治疗的淋巴瘤患者中STAT3和STAT5磷酸化的降低。因此，我们描述了一种用于确定临床标本中癌蛋白表达和磷酸化的新的高灵敏度方法，以开发新的癌症治疗方法。
