原代成纤维细胞和纤维肉瘤细胞自分泌转化生长因子β1浓度检测及对转化生长因子β1增殖的调节

Detection of autocrine transforming growth factor-beta 1 concentration in the primary fibroblasts and fibrosarcoma cells and effects on cell proliferation

背景:体外实验常采用外源性转化生长因子β1刺激来观察细胞增殖变化,易忽略细胞本身自分泌转化生长因子β1的影响。目的:检测原代成纤维细胞和纤维肉瘤细胞(L929)自分泌转化生长因子β1的浓度和细胞增殖的变化。方法:Elisa试剂盒检测原代成纤维细胞和纤维肉瘤细胞各时相点细胞内和培养上清中生长转化因子β1水平。结果与结论:纤维肉瘤细胞内转化生长因子β1浓度随培养时间延长而升高,原代成纤维细胞内转化生长因子β1浓度随培养时间延长而降低;两种细胞培养上清转化生长因子β1浓度均随时间而上升,但纤维肉瘤细胞显著高于原代成纤维细胞(P<0.01),峰值时达到原代成纤维细胞的20倍;两种细胞增殖曲线在72h内均随时间而显著增高(P<0.01),同一时间点比较,纤维肉瘤细胞明显高于原代成纤维细胞。提示自分泌转化生长因子β1具有促进细胞增殖的作用,不同的自分泌水平可能是影响原代成纤维细胞这种正常细胞和纤维肉瘤细胞肿瘤细胞对于外源转化生长因子β1反应不同的重要原因。

BACKGROUND：Although tremendous strides have been made in clarifying the different regulation mechanisms of cell proliferation on normal cells and tumor cells,until now,it is still not clear.Exogenous stimulation of transforming growth factor-β1（TGF-β1） is often used in vitro experiments to observe the changes in cell proliferation and the effects of autocrine TGF-β1 itself are easily ignored.OBJECTIVE：To detect the concentration of autocrine TGF-β1and cell proliferation of the primary fibroblasts（Fb） and fibrosarcoma cells（L929）,and to investigate the effects of the autocrine TGF-β1 on cell proliferation.METHODS：The TGF-β1 levels in cells and supernatants of Fb and L929 cells were detected by ELISA test kit.The proliferation of Fb and L929 cells were detected by MTT assay.RESULTS：TGF-β1 concentration in L929 cells increased with prolonged incubation time,whereas TGF-β1 concentration in Fb cells decreased with prolonged incubation time.TGF-β1 concentration in supernatant of Fb and L929 cells increased with prolonged incubation time,but TGF-β1 concentration in supernatant of L929 cells was significantly higher than that of Fb cells（P 0.01）.The cell proliferation curves both in Fb and L929 cells were significantly increased within 72 hours（P 0.01）.Compared with the same time,the cell proliferation of L929 cells was significantly higher than that of Fb cells.The autocrine TGF-β1 of the Fb and L929 cells can promote cell proliferation,and different levels of autocrine TGF-β1 maybe the important reason for different reactions of Fb and L929 cells stimulated by exogenous TGF-β1.