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[目的]采用细胞培养的方法研究核黄素对人食管癌细胞株生长增殖的影响,探讨核黄素营养水平与食管癌的关系。[方法]常规培养人食管癌细胞株Eca109细胞。(1)用不同浓度的核黄素作用于细胞株,MTT比色法测定Eca109细胞的增殖活性,计算细胞增殖率。(2)采用流式细胞仪法测定核黄素作用Eca109细胞后细胞周期的改变。(3)免疫组织化学法测定核黄素对Eca109细胞作用24 h Cyclin D1蛋白表达的影响。(4)HE染色以观察核黄素作用24 h对Eca109细胞的分化影响。[结果](1)不同浓度的核黄素作用于Eca109细胞后,其增殖率未见明显变化。(2)核黄素的加入使Eca109的细胞周期改变,出现G2/M期阻滞,但不促进其凋亡。(3)核黄素处理24 h后,经过免疫组化测定,食管癌细胞Cyclin D1蛋白表达无降低,与阴、阳性对照组相比差异均无统计学意义(P﹥0.05)。(4)HE染色观察各浓度组细胞形态形态无明显改变。[结论]核黄素不会影响食管癌Eca109的增殖,但可能将细胞阻滞在M期。
[Objective] The research aimed to study the effect of riboflavin on the growth and proliferation of human esophageal cancer cell lines by cell culture and to explore the relationship between riboflavin nutrition and esophageal cancer. [Method] The human esophageal cancer cell line Eca109 cells were routinely cultured. (1) Different concentrations of riboflavin act on the cell line, MTT colorimetric assay of Eca109 cell proliferation activity, calculate the cell proliferation rate. (2) The cell cycle changes of Eca109 cells treated with riboflavin were determined by flow cytometry. (3) The effect of riboflavin on the expression of Cyclin D1 protein in Eca109 cells by immunohistochemical method for 24 h. (4) HE staining to observe the differentiation of Eca109 cells induced by riboflavin for 24 h. [Results] (1) The proliferation rate of Eca109 cells treated with different concentrations of riboflavin did not change significantly. (2) The addition of riboflavin changed the cell cycle of Eca109, which resulted in G2 / M arrest, but not apoptosis. (3) After 24 h of riboflavin treatment, the expression of Cyclin D1 protein in esophageal cancer cells was not decreased after immunohistochemical analysis, and there was no significant difference between the control group and the negative control group (P> 0.05). (4) HE staining showed no significant changes in cell morphology and morphology at each concentration. [Conclusion] Riboflavin does not affect the proliferation of esophageal cancer Eca109 cells, but may arrest the cells in M phase.