Screening of differential proteins in lung adenocarcinoma and squamous cell carcinoma by laser capture microdissection combined with protein microarray

Release date: 2007-09-10

Screening of differentially expressed proteins in lung adenocarcinoma and squamous cell carcinoma by laser capture microdissection combined with protein microarray. Second Affiliated Hospital of Xi'an Jiaotong University School of Medicine, Tian Yingxuan, Yang Yuying, Nan Yandong, Zhang Wei, Zhou Bin, Bo Lina, Huo Shufen, Institute of Oncology, Second Hospital of Zhejiang University School of Medicine, Yu Jiekai Zhengshu lung squamous cell carcinoma and adenocarcinoma are the main tissue types of lung cancer. The use of proteomics methods to compare the differential proteins of two tumors is an effective means to elucidate its cancerous mechanism, screening and diagnostic markers. However, the heterogeneity of tumor tissue is a major problem in proteomics research. Laser capture microdissection (LCM) technology is an effective method to solve the problem of tumor tissue heterogeneity. Its combined surface enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) protein chip technology allows screening of differential proteins from trace samples. We used LCM in combination with SELDI-TOF-MS protein chip technology and support vector machine (SVM) method for the feasibility of differential proteomic studies of lung adenocarcinoma and squamous cell carcinoma. 6 cases of fresh lung adenocarcinoma and 7 cases of lung squamous cell carcinoma were prepared for 8um thickness frozen sections; modified HE staining; 1.2×105/one homogenous adenocarcinoma cells and 1.45×105/squamous carcinoma cells were selectively obtained by LCM. The protein expression profiles of adenocarcinoma and squamous carcinoma cells were analyzed by PBS II+ SELDI-TOF-MS analyzer (IMAC chip) and the differences were compared. The discriminative potency of candidate marker proteins was screened and verified using the ZUCI-ProteinChip DataAnalyze System software package. The results showed that 87 differential protein peaks were screened in SELDI spectra between squamous cell carcinoma and adenocarcinoma cells. The 10 protein peaks with the most significant differences were used as potential candidate marker proteins. Four proteins (molecular weights of 2505 U, 4004 U, 4847 U, 11412 U) were highly expressed in squamous cell carcinoma; six proteins (molecular weights of 3333 U, 3592 U, 3848 U, 5036 U, 5191 U, 5211, respectively) U) is lowly expressed in squamous cell carcinoma and highly expressed in adenocarcinoma. The expression of 4847 U in squamous cell carcinoma and adenocarcinoma was statistically significant (P<0.05). 10 differential proteins were randomly combined to form a combined model; the SVM and ROC curves were used to establish a classification prediction model and the efficacy of each model was evaluated; the scores composed of 3 protein spots (molecular weight 4847 U, 11412 U, 3592 U) were selected. Type diagnostic model. The blinded cross-validation model can accurately distinguish between squamous cell carcinoma and adenocarcinoma, and its sensitivity and specificity can reach 100%. It is suggested that LCM combined with SELDI protein chip technology can screen sensitive and specific lung cancer marker proteins and typing diagnostic models; this technology will provide a new and effective tool for early diagnosis of lung adenocarcinoma. ——Midi Medical Network

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