应用SELDI-TOF-MS技术筛选TGF-β1刺激胃癌细胞分泌蛋白的研究

Study on screening protein secretion after TGF-β1 stimulation of gastric cancer cells by SELDI-TOF-MS technique

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 SELDI-TOF-MS technology was used to contrastly analyse the changes of protein expression profiles of gastric cancer cells after TGF- β1 stimulation, and provide theoretical and experimental foundation for screening different significant proteins. Methods Serial suheuhivation gastric cancer cells BGC823 , MKN45 , and SGC7901 cultured in vitro were grouped into test and control groups. TGF-β1 was added to the test group, but not to control group. Cell culture fluid was collected and centrifuged after cultured 24h, and crossing with WCX2 protein chip to detect protein differences. Results When the test group was compared with control group, we found： （ 1 ） thirteen different proteins in BGC823 cells after TGF-β1 stimulaton, and their M/Z were M4294,M4932,M4945,M4972, M4991, M5015, 5036, M5060, M5153, M5180, M5197, M8577, and M8784, respectively; （2） eighteen different proteins in MKN45 cells after TGF-β1 stimulaton, and their M/Z were M4292, M4931, M4945, M4972, M4990, M5014, M5152, M5178, M7055, M8190, M8570, M8652, M8670, M8780, M9963, M10098 , M10523 , and M11653 , respectively; （ 3 ） eight difference proteins in SGC7901 cells after TGF-β1 stimulation, and their M/Z were M4945, M4972, M4992, M5015, M5180, M7056, M8573, and M8604, respectively. By comparing three protein expression profiles of gastric cancer cells after TGF-β1 treatment, we found two significcant proteins with common differences, that had M/Z of M4945 and M4972, respectively. Conclusions The biological markers whose M/Z is M4945, M4972 with gastric cancer characteristic and associated with TGF-β1 have been screened, which can be as the basis for early prediction and clinical diagnosis research on metastasis and invasiveness of gastric cancer.