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Establishment of hypoxia-resistant gastric cancer cell line and biological characteristics identification |
CHE Xiaofang1,2 SHI Xiaonan1,2 ZHENG Chunlei1,2 YANG Zichang1,2 ZHANG Xiaojie1,2 QU Xiujuan1,2 LIU Yunpeng1,2 |
1.Department of Medical Oncology, the First Hospital of China Medical University, Liaoning Province, Shenyang 110001, China;
2.Key Laboratory of Anticancer Drugs and Biotherapy of Liaoning Province, Liaoning Province, Shenyang 110001, China |
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Abstract Objective To establish a hypoxic-resistant gastric cancer cell line and analyze its biological characteristics to provide a good model for the study of hypoxic microenvironment in gastric cancer. Methods The hypoxic-resistant gastric cancer cell lines MGC803/hypo and BGC823/hypo, which were tolerant to 2% O2, were established by gradually decreasing the oxygen concentration. The morphology of the cells was observed by inverted microscope. The cell metastasis ability was detected by Transwell chamber. The cell proliferation ability was detected by MTT assay. The protein expression was detected by Western blot. The biological characteristics of the gastric cancer cell lines were compared with those of their parents. Results Compared with the parental gastric cancer cell line, the hypoxic-resistant gastric cancer cell line showed spindle-shaped changes and loosely arranged. The color of the culture medium was easy to turn yellow, the migration ability was significantly enhanced (P < 0.05), and the proliferation ability was weakened (P > 0.05). Hypoxia-related index HIF-2α was significantly up-regulated, Glut1 was up-regulated in glycolytic index, ZO-1 was down-regulated in epithelial mesenchymal transition (EMT), vimentin was up-regulated, and metastasis-related indicators MUC1, integrin α5, and integrin β1 were up-regulated. Conclusion This study established hypoxic-resistant gastric cancer cell lines MGC803/hypo and BGC823/hypo which can tolerate 2% O2 concentration. The Metastasis ability is obviously enhanced and the proliferative ability is relatively weakened. This provides a good model for exploring the mechanism of hypoxic microenvironment promoting tumor metastasis.
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