3种增塑剂单独和联合暴露对MCF-7细胞增殖的影响

Study on the combined effects of the three kinds of plasticizers on the proliferations of MCF-7 cells

为了探究结构差异较大、应用较为广泛的几类增塑剂雌激素活性的联合效应,选择邻苯二甲酸二丁酯(DBP)、双酚A(BPA)和壬基酚(NP)作为邻苯二甲酸酯、双酚A类和烷基酚类增塑剂的代表物进行试验。用DBP、BPA和NP单独及两两混合处理MCF-7细胞。采用M丌法检测培养24 h、48 h、72 h和96 h时的细胞增殖情况。采用流式细胞术检测药物培养48 h后的细胞生长周期分布,并计算细胞增殖指数(PI)。运用效应叠加模型(ES)判定联合效应类型。结果表明,在单独暴露试验中,DBP、BPA和NP组PI均大于1,且均能提高S期(DNA合成期)细胞比例。因此,DBP、BPA和NP均能显著促进MCF-7细胞增殖。混合暴露试验中,1)DBP和BPA在MTT试验中24 h、48h、72 h和96 h时的效应叠加指数(ESI)分别为1.013 9、1.023 8、0.9999、1.010 8,在流式细胞仪试验中ESI为1.014 1。因此,DBP和BPA的雌激素活性联合效应为加和作用。2)DBP和NP在MTT试验中24 h、48 h、72 h和96 h时的ESI分别为1.004 0、1.008 6、1.011 5、1.0103,流式细胞仪试验中ESI为0.997 0。因此,DBP和NP的雌激素活性联合效应为加和作用。3)BAP和NP在MTT试验中24 h、48 h、72 h和96 h时的ESI分别为0.980 6、0.981 8、0.977 7、0.973 3,流式细胞仪试验的ESI为0.912 8。由此可知,BPA和NP的雌激素活性联合效应为拮抗作用。因此,可以采用MCF-7细胞增殖试验研究环境污染物雌激素活性联合效应。

The aim of the present study is to make an exploration of the combined effects of the various plasticizers which have broadly been met with but differ largely in structure.In this paper,dibutyl phthalate（DBP）,bisphenol A（BPA） and nonylphenol（NP） were used to represent phthalic acid esters,bisphenol A analogues and alkylphenol.MCF-7 cells were grown on media containing one or two of these drugs.Furthermore,the proliferations of the cells cultured for 24 h,48 h,72 h and 96 h were tested by MTT assay and the cell proliferation indices（PI） were calculated.The combined effect of plasticizers could be identified and determined by a summation model,whereas all of the PIs in the DBP,BPA and NP groups could be tested in more than one single dose experiments,which help to increase the proportion of cells in the S phase.Therefore,the result displayed that DBP,BPA and NP could promote the proliferation of MCF-7 cells remarkably.Moreover,in the MTT experiment,the ESIs of DBP and BPA had been made known equal to 1.013 9,1.023 8,0.999 9 and 1.010 8 respectively as a result of 24 h,48 h,72 h and 96 h experiment,whereas that of the ESI turned out to be 1.014 1 in the flowing cytometry experiments.Thus,the estrogenic activity excited by the combined effects of DBP and BPA proved to be additive in effect.The ESIs of DBP and NP as the result of the MTT experiments of 24 h,48 h,72 h and 96 h later had been found equal to 1.0040,1.008 6,1.011 5 and 1.010 3,respectively.Meanwhile,the effect of ESI came out at 0.997 0 in the flow cytometry experiment.Hence the combined effects of DBP and NP turned out to be additive.As for the MTT experiments,the ESIs of BPA and NP had been worked out equal to 0.980 6,0.981 8,0.973 3 and0.977 7 correspondingly for different time intervals.Thus,the estrogen activity of BPA and DBP indicated a mild antagonism for the ESI was equal to 0.9128 asa result of the flow cytometry.Furthermore,the developed method proved to be suitable for screening the combined effect of the various estrogenic substances along with its underlying mechanism.