Study of differential proteins in lung adenocarcinoma using laser capture microdissection combined with liquid chip-mass spectrometry technology  

作　　者：BU Li-na YANG Shuan-ying LI Feng-tao SHANG Wen-li ZHANG Wei HUO Shu-fen NAN Yan-dong TIAN Ying-xuan DU Jie LIN Xiu-li LIU Yan-feng LIN Yu-rong RONG Biao-xue 

机构地区：[1]Department of Respiratory Medicine, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, Shaanxi 710004, China [2]Department of General Thoracic Surgery , Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an, Shaanxi 710004, China [3]Department of Respiratory Medicine, Xi'an Second Hospital, Xi'an, Shaanxi 710004, China

出　　处：《Chinese Medical Journal》2010年第22期3309-3313,共5页中华医学杂志（英文版）

基　　金：This work was supported by grants from the National Natural Science Foundation of China （No. 30570795） and Program for New Century Excellent Talents in University （No. NECT-06-0845） and the Program in Science and Technology of Xi＇an, Shaanxi Province （No. SF08009（1））.

摘　　要：Background In recent years the proportion of lung adenocarcinoma （adCA） which occurs in lung cancer patients has increased. Using laser capture microdissection （LCM） combined with liquid chip-mass spectrometry technology, we aimed to screen lung cancer biomarkers by studying the proteins in the tissues of adCA. Methods We used LCM and magnetic bead based weak cation exchange （MB-WCX） to separate and purify the homogeneous adCA cells and normal cells from six cases of fresh adCA and matched normal lung tissues. The proteins were analyzed and identified by matrix assisted laser desorption/ionization time-of-fight mass spectrometry （MALDI-OF-MS）. We screened for the best pattern using a radial basic function neural network algorithm. Results About 2.895x10s and 1.584x10s cells were satisfactorily obtained by LCM from six cases of fresh lung adCA and matched normal lung tissues, respectively. The homogeneities of cell population were estimated to be over 95% as determined by microscopic visualization. Comparing the differentially expressed proteins between the lung adCA and the matched normal lung group, 221 and 239 protein peaks, respectively, were found in the mass-to-charge ration （M/Z） between 800 Da and 10 000 Da. According to ttest, the expression of two protein peaks at 7521.5 M/Zand 5079.3 M/Z had the largest difference between tissues. They were more weakly expressed in the lung adCA compared to the matched normal group. The two protein peaks could accurately separate the lung adCA from the matched normal lung group by the sample distribution chart. A discriminatory pattern which can separate the lung adCA from the matched normal lung tissue consisting of three proteins at 3358.1 M/Z, 5079.3 M/Z and 7521.5 M/Z was established by a radial basic function neural network algorithm with a sensitivity of 100% and a specificity of 100%. Conclusions Differential proteins in lung adCA were screened using LCM combined with liquid chip-mass spectrometry technology, and a biomarker model was establisBackground In recent years the proportion of lung adenocarcinoma （adCA） which occurs in lung cancer patients has increased. Using laser capture microdissection （LCM） combined with liquid chip-mass spectrometry technology, we aimed to screen lung cancer biomarkers by studying the proteins in the tissues of adCA. Methods We used LCM and magnetic bead based weak cation exchange （MB-WCX） to separate and purify the homogeneous adCA cells and normal cells from six cases of fresh adCA and matched normal lung tissues. The proteins were analyzed and identified by matrix assisted laser desorption/ionization time-of-fight mass spectrometry （MALDI-OF-MS）. We screened for the best pattern using a radial basic function neural network algorithm. Results About 2.895x10s and 1.584x10s cells were satisfactorily obtained by LCM from six cases of fresh lung adCA and matched normal lung tissues, respectively. The homogeneities of cell population were estimated to be over 95% as determined by microscopic visualization. Comparing the differentially expressed proteins between the lung adCA and the matched normal lung group, 221 and 239 protein peaks, respectively, were found in the mass-to-charge ration （M/Z） between 800 Da and 10 000 Da. According to ttest, the expression of two protein peaks at 7521.5 M/Zand 5079.3 M/Z had the largest difference between tissues. They were more weakly expressed in the lung adCA compared to the matched normal group. The two protein peaks could accurately separate the lung adCA from the matched normal lung group by the sample distribution chart. A discriminatory pattern which can separate the lung adCA from the matched normal lung tissue consisting of three proteins at 3358.1 M/Z, 5079.3 M/Z and 7521.5 M/Z was established by a radial basic function neural network algorithm with a sensitivity of 100% and a specificity of 100%. Conclusions Differential proteins in lung adCA were screened using LCM combined with liquid chip-mass spectrometry technology, and a biomarker model was establis

关 键 词：lung adenocarcinoma laser capture microdissection magnetic beads mass spectrometry 

分 类 号：R686[医药卫生—骨科学]