Hunt Allison L, Pierobon Mariaelena, Baldelli Elisa, Oliver Julie, Mitchell Dave, Gist Glenn, Bateman Nicholas W, Larry Maxwell G, Petricoin Emanuel F, Conrads Thomas P
1Women's Service Line, Inova Health System, 3300 Gallows Rd., Falls Church, VA 22042 USA.
2Gynecologic Cancer Center of Excellence, Department of Obstetrics and Gynecology, Uniformed Services University and Walter Reed National Military Medical Center, 8901 Wisconsin Avenue, Bethesda, MD 20889 USA.
Clin Proteomics. 2020 Mar 9;17:9. doi: 10.1186/s12014-020-09272-z. eCollection 2020.
Reversible protein phosphorylation represents a key mechanism by which signals are transduced in eukaryotic cells. Dysregulated phosphorylation is also a hallmark of carcinogenesis and represents key drug targets in the precision medicine space. Thus, methods that preserve phosphoprotein integrity in the context of clinical tissue analyses are crucially important in cancer research. Here we investigated the impact of UV laser microdissection (UV LMD) and IR laser capture microdissection (IR LCM) on phosphoprotein abundance of key cancer signaling protein targets assessed by reverse-phase protein microarray (RPPA). Tumor epithelial cells from consecutive thin sections obtained from four high-grade serous ovarian cancers were harvested using either UV LMD or IR LCM methods. Phosphoprotein abundances for ten phosphoproteins that represent important drug targets were assessed by RPPA and revealed no significant differences in phosphoprotein integrity from those obtained using higher-energy UV versus the lower-energy IR laser methods.
可逆性蛋白质磷酸化是真核细胞中信号转导的关键机制。磷酸化失调也是致癌作用的一个标志，并且是精准医学领域的关键药物靶点。因此，在临床组织分析中能够保持磷蛋白完整性的方法在癌症研究中至关重要。在此，我们研究了紫外激光显微切割（UV LMD）和红外激光捕获显微切割（IR LCM）对通过反相蛋白质微阵列（RPPA）评估的关键癌症信号蛋白靶点的磷蛋白丰度的影响。使用UV LMD或IR LCM方法从取自4例高级别浆液性卵巢癌的连续薄切片中获取肿瘤上皮细胞。通过RPPA评估了代表重要药物靶点的10种磷蛋白的磷蛋白丰度，结果显示，与使用高能量紫外激光方法相比，低能量红外激光方法所获得的磷蛋白完整性没有显著差异。
