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【目的】观察血瘀证食管鳞癌患者血清对食管鳞癌EC9706细胞增殖和细胞周期的影响,探讨血瘀证食管鳞癌患者血液微环境对食管癌细胞的作用。【方法】将EC9706细胞以含胎牛血清(FCS)的RPMI-1640培养液置于37℃、体积分数5%饱和湿度的CO2培养箱中孵育24 h,再以不含血清的培养液孵育24 h后,分别加入血瘀证、脾气虚证食管鳞癌患者血清和健康人血清,四甲基偶氮唑盐(MTT)法检测细胞增殖情况,光镜下观察细胞形态变化,流式细胞术观察细胞周期变化。【结果】血瘀证、脾气虚证食管鳞癌患者血清刺激细胞增殖率为50%时的浓度(PI50)分别为71.1μL/m L和118μL/m L;2组人血清刺激细胞增殖的峰值均出现在时间点48 h。光镜观察结果显示,血瘀证组、脾气虚证组血清刺激细胞呈长梭形或多角形改变,血瘀证组细胞数目较脾虚证组增多。细胞周期结果显示,与脾气虚组比较,血瘀证组患者血清刺激的G1增殖期细胞百分率明显减少,S增殖期细胞百分率明显增高(均P<0.05)。【结论】血瘀证食管鳞癌患者血清微环境较脾气虚证食管鳞癌患者更有利于食管癌细胞的增殖,其作用机制可能与调节细胞周期有关。
【Objective】 To observe the effect of serum of patients with esophageal squamous cell carcinoma of blood stasis on the proliferation and cell cycle of esophageal squamous cell carcinoma EC9706 cells and to investigate the effect of blood microenvironment on esophageal cancer cells in patients with esophageal squamous cell carcinoma of blood stasis. 【Method】 EC9706 cells were cultured in RPMI-1640 medium containing fetal bovine serum (FCS) for 24 h in a CO2 incubator with a volume fraction of 5% saturated humidity at 37 ° C., and incubated with serum-free medium for 24 h h, were added to the blood stasis, spleen qi deficiency of esophageal squamous cell carcinoma serum and healthy human serum, tetramethylzole (MTT) method to detect cell proliferation, morphological changes observed under light microscope, flow cytometry Observation of cell cycle changes. 【Results】 The results showed that the PI50 of serum-stimulated cell proliferation rate was 71.1 μL / m L and 118 μL / m L respectively for blood-stasis syndrome and spleen-qi deficiency syndrome group. The peak of cell proliferation stimulated by two groups of human serum All appeared at the time point 48 h. Light microscopy results showed that the serum-stimulated cells in blood stasis syndrome group and spleen qi deficiency syndrome group showed long fusiform or polygonal changes, and the number of cells in blood stasis syndrome group was more than that in spleen deficiency syndrome group. The results of cell cycle showed that the percentage of cells in G1 proliferative phase and the percentage of cells in S proliferative phase were significantly increased (P <0.05). 【Conclusion】 The serum microenvironment of patients with esophageal squamous cell carcinoma of blood stasis is more beneficial to the esophageal cancer cell proliferation than the patients with esophageal squamous cell carcinoma of the spleen qi deficiency syndrome. The possible mechanism may be related to the regulation of cell cycle.