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目的应用SELDI-TOF-MS技术对比分析胃癌细胞经转移生长因子β1(TGF-β1)刺激分泌后的蛋白质谱变化,为下一步筛选有意义的差异蛋白奠定理论和实验基础。方法体外传代培养胃癌细胞株SGC7901,BGC823,MKN45,并分为实验组和对照组。实验组加入TGF-β1刺激,而对照组不加入TGF-β1刺激。培养24h后收集细胞培养液并离心,与WCX2蛋白芯片杂交后,上机进行SELDI-TOF-MS检测。结果(1)BGC823细胞受TGF-β1刺激后,与对照组比较发现了13个差异蛋白峰,质荷比(M/Z)是M4294,M4932,M4945,M4972,M4991,M5015,M5036,M5060,M5153,M5180,M5197,M8577,M8784。(2)MKN45细胞受TGF-β1刺激后,与对照组比较发现了18个差异蛋白峰,M/Z分别是M4292,M4931,M4945,M4972,M4990,M5014,M5152,M5178,M7055,M8190,M8570,M8652,M8670,M8780,M9963,M10098,M10523,M11653。(3)SGC7901细胞受TGF-β1刺激后,与对照组比较发现了8个有意义的差异蛋白峰,M/Z分别是M4945,M4972,M4992,M5015,M5180,M7056,M8573,M8604。(4)比较3种胃癌细胞经TGF-β1刺激后的蛋白质图谱,共发现2个有意义的共同差异蛋白峰,它们的M/Z分别是M4945,M4972。结论筛选出与TGF-β1作用相关的胃癌特征性的生物标记物——M4945,M4972,这些生物标记为胃癌患者浸润、转移的早期预测和临床诊断的进一步研究奠定了基础。
Objective To compare and analyze the changes of protein profiles in gastric cancer cells stimulated by TGF-β1 using SELDI-TOF-MS technology, and lay the theoretical and experimental foundation for further screening of differentially expressed proteins. Methods Gastric cancer cell lines SGC7901, BGC823 and MKN45 were subcultured and divided into experimental group and control group. The experimental group added TGF-β1 stimulation, while the control group did not add TGF-β1 stimulation. After cultured for 24h, the cell culture medium was collected and centrifuged. After hybridization with the WCX2 protein chip, SELDI-TOF-MS was performed on the machine. Results (1) Thirteen differential protein peaks were found in BGC823 cells stimulated with TGF-β1 compared with the control group. The mass-to-charge ratio (M / Z) of BGC823 cells was M4294, M4932, M4945, M4972, M4991, M5015, M5036, M5060, M5153, M5180, M5197, M8577, M8784. (2) 18 different protein peaks were found in MKN45 cells stimulated by TGF-β1 compared with the control group, and M / Z were M4292, M4931, M4945, M4972, M4990, M5014, M5152, M5178, M7055, M8190, M8570 , M8652, M8670, M8780, M9963, M10098, M10523, M11653. (3) There were eight significant differential protein peaks in SGC7901 cells stimulated by TGF-β1 compared with the control group. M / Z were M4945, M4972, M4992, M5015, M5180, M7056, M8573 and M8604, respectively. (4) Comparison of the protein profiles of the three kinds of gastric cancer cells stimulated by TGF-β1, we found two significant common difference protein peaks, their M / Z are M4945, M4972 respectively. Conclusion The biomarkers M4945 and M4972, which are characteristic of gastric cancer, were selected according to the role of TGF-β1. These biomarkers have laid the foundation for the further study on the early prediction and clinical diagnosis of gastric cancer.