山东医药2017，Vol.57Issue(6)：1-4,4.DOI:10.3969/j.issn.1002-266X.2017.06.001
低氘水对人肺癌细胞 A549增殖和分化的影响及机制
Effects of deuterium-depleted water on proliferation and differentiation of lung cancer A549 cells
摘要
Abstract
Objective To study the effects of deuterium-depleted water ( DDW) on human lung cancer cell line A549 and to explore the underlying molecular mechanisms .Methods A549 cells were randomly divided into two groups:the observation group and the control group , which were seperately cultured in the medium prepared with 50 ppm DDW and in the medium prepared with normal water for 30 d.The morphology of cells was observed under an inverted microscope . Cell growth was measured using the colorimetric cell counting kit-8 ( CCK-8) reagent to compare the growth rate of cells be-tween the two groups .The cells in the two groups were synchronized by serum starvation and then were separately stimula -ted with medium containing 10%serum of DDW and medium containing normal water .At 36 and 48 h after the stimula-tion, cells from each group were trypsinized , stained with propidium iodide ( PI) and then were analyzed by fluorescence-activated cell sorting (FACS) to determine the percentages of cells distributed in G 1, S and G2/M phases during cell cy-cle.Total proteins were extracted from cells cultured in normal medium or DDW medium .Western blotting was conducted to detect the expression levels of MEK 1 and EGFR, phosphorylation of serine 217 and serine 298 on MEK1, and phospho-rylation of tyrosine 1016 and tyrosine 1110 .The expression levels of several alveolus-specific protein markers ( type Ⅰalveolus markers:AQP5 and T1a, typeⅡalveolus markers SPB, SPC1, SPC2 and CFTR) in cells of the two groups were also compared.Results In the control group, the cells were cuboidal and in close contact with accumulation of cells .In the observation group , the cells were spindle , and intercellular contact was loose with cell elongation .The OD of the obser-vation group was lower than that of the control group (all P<0.05).After the serum starvation, cells were stimulated for 36 h, the percentage of cells in G 1 phase of the observation group was 71.75%, which was higher than that of the control group (57.01%), the percentage of cells in S phase was 22.43%, which was lower than that of the control group (35.88%), (all P<0.05).There was no significant difference in cell cycle ratio between the two groups at 48 h.No sig-nificant difference was found in the expression of MEK 1, EGFR and the phosphorylation of EGFR on tyrosine 1016 and 1110 between these two groups .In the observation group , the phosphorylation of MEK 1 on both serine 217 and serine 298 was significantly decreased (all P<0.01).The expression level of SPC2 in the observation group was significantly in-creased as compared with that of the control group (P<0.01).Conclusions Long-term cultivation of A549 lung cancer cells with DDW medium significantly inhibits the cell proliferation and differentiation , which is related to decreasing phos-phorylation level of MEK1, arresting the G1/S phase transformation, inducing A549 cells from type Ⅱ alveolar epithelial cells to type I alveolar epithelial cells and improving the differentiation of A 549 cells.关键词
肺癌/低氘水/细胞周期/丝裂原活化的细胞外信号调节激酶/细胞分化Key words
lung carcinoma/deuterium-depleted water/cell cycle/mitogen-activated extracellular signal-regulated kinase/cell differentiation分类
医药卫生引用本文复制引用
戚冰雪,李子宜,丛峰松..低氘水对人肺癌细胞 A549增殖和分化的影响及机制[J].山东医药,2017,57(6):1-4,4.基金项目
国家自然科学基金资助项目（81272478）；教育部重点实验室开放课题（AF4150013）。 （）
