Differentially expressed proteins in the human esophageal cancer cell line Eca-109, in the presence and absence of gemcitabine

作者：Ma, Zenghuang; Xue, Xiaojie^*

来源：Molecular Medicine Reports, 2018, 17(1): 1873-1878.

DOI：10.3892/mmr.2017.8041

• gemcitabine
• esophageal cancer
• differential proteins
• western blotting
• matrix-assisted laser desorption
• ionization time of flight mass spectrometry

The present study aimed to screen and study the roles of differentially expressed proteins in the human esophageal cancer cell line Eca-109, in the presence and absence of gemcitabine (GEM). The 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di-phenytetrazoliumromide (MTT) method was used to assay the vitality of the Eca-109 cells following treatment with GEM (1-16 mu g/ml). The cell apoptosis was measured by using fluorescence activated cell sorting. The proteins in the treated Eca-109 cells were extracted, validated, and assayed via two-dimensional gel electrophoresis combined with matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The differentially expressed proteins were then determined by western blotting. Furthermore, alterations in mitochondrial ultrastructure of the treated cells were observed under a transmission electron microscope. GEM significantly inhibited the growth of the Eca-109 cells in a concentration- and time-dependent manner, and the 50% inhibition concentration (IC50) value was 3.87 mu g/ml. The MALDI-TOF-MS analysis revealed that there were three differentially expressed proteins following the GEM treatment, compared with the control. The differential proteins were verified to be B cell lymphoma-2 associated X, apoptosis regulator (Bax)-, apoptosis-associated speck-like protein containing a CARD (ASC) and myeloid cell leukemia sequence (Mcl)-1. Western blotting revealed that the expression levels of ASC and Bax- proteins in the treated cancer cells were significantly upregulated, whereas the Mcl-1 protein expression was markedly downregulated compared with the control. Furthermore, the GEM treatment destroyed the mitochondrial ultrastructure of the cancer cells, leaving swelled mitochondria, a fading matrix and destroyed the mitochondrial cristae. GEM significantly inhibits the growth and promotes apoptosis of the Eca-109 cells, due to the alterations in the expression levels of the differential proteins, including ASC, Mcl-1 and Bax-.

• 出版日期2018-1
• 单位湖北理工学院