Li Xiuling, Zheng Shutao, Liu Qing, Liu Tao, Liang Meng, Gao Xiangpeng, Lu Mang, Sheyhidin Ilyar, Lu Xiaomei
Clinical Medical Research Institute, First Affiliated Hospital of Xinjiang Medical University, Xinjiang Uygur Autonomous Region, Urumqi, P.R. China.
State Key Lab Incubation Base of Xinjiang Major Diseases Research, First Affiliated Hospital of Xinjiang Medical University, Xinjiang Uygur Autonomous Region, Urumqi, P.R. China.
Mol Carcinog. 2015 Sep;54(9):779-88. doi: 10.1002/mc.22145. Epub 2014 Mar 26.
The aim of the study was to identify candidate biomarkers for esophageal squamous cell carcinoma (ESCC) in Kazakh ethnic in Xinjiang as well as to reveal the potential role of Annexin A2 in ESCC carcinogenesis and progression. Five paired of Kazakh's ESCC tissues (T) and matched adjacent morphologically normal tissues (N) were separated by two-dimensional electrophoresis (2-DE) and differential proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Annexin A2 was identified as a down-regulated protein in Kazakh's ESCC and further validated by immunohistochemistry (IHC) in 77 Kazakh's ESCC formalin-fixed paraffin-embeded (FFPE) samples. The expression level of Annexin A2 protein significantly correlated with the degree of ESCC differentiation and depth of invasion. For clarification of the role of Annexin A2 in regulating cell phenotype, in vitro eukaryotic expression vectors harboring full length Annexin A2 (pCMV-XL5-Annexin A2) was tranfected into Eca109 cells, and transfection effects were evaluated by RT-PCR and Western blotting analysis, respectively. Functionally, there was a significant decrease in cell proliferation, migration, and invasion capability in Eca109 with transfected pCMV-XL5-Annexin A2 compared to the controls. Furthermore, up-regulating Annexin A2 can significantly cause cell cycle arrest at the G2 phase, but no apoptosis was induced. Together, our findings suggested that Annexin A2 was involved in malignant phenotype and was a potential biomark for molecular classification in ESCC.
本研究旨在寻找新疆哈萨克族食管鳞状细胞癌（ESCC）的候选生物标志物，并揭示膜联蛋白A2在ESCC发生发展中的潜在作用。采用二维电泳（2-DE）技术分离5对哈萨克族ESCC组织（T）及其配对的形态学正常的癌旁组织（N），并用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）鉴定差异蛋白。膜联蛋白A2被鉴定为哈萨克族ESCC中下调的蛋白，并通过免疫组织化学（IHC）在77例哈萨克族ESCC福尔马林固定石蜡包埋（FFPE）样本中进一步验证。膜联蛋白A2蛋白的表达水平与ESCC的分化程度和浸润深度显著相关。为阐明膜联蛋白A2在调节细胞表型中的作用，将携带全长膜联蛋白A2的真核表达载体（pCMV-XL5-膜联蛋白A2）转染至Eca109细胞，分别通过RT-PCR和蛋白质印迹分析评估转染效果。在功能上，与对照组相比，转染pCMV-XL5-膜联蛋白A2的Eca109细胞的增殖、迁移和侵袭能力显著降低。此外，上调膜联蛋白A2可显著导致细胞周期阻滞在G2期，但未诱导细胞凋亡。总之，我们的研究结果表明膜联蛋白A2参与了恶性表型的形成，是ESCC分子分类的潜在生物标志物。
