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目的研究不同试验条件对MCF-7增殖实验检测雌激素效应的影响。方法选用两种来源(ATCC和某肿瘤医院)的MCF-7细胞,研究不同浓度、不同培养时间、不同溶剂、不同培养液条件下MCF-7细胞对17β-雌二醇(E2)、乙炔雌二醇(EE2)、雌三醇(E3)、雌酮(E1)的反应,以优化MCF-7增殖实验条件。结果不同E2浓度条件下源于ATCC的MCF-7细胞增殖率高于肿瘤医院提供的MCF-7细胞;随着E2浓度的增加,细胞增殖率增加,当E2浓度达到10-9 mol/L时,细胞增殖率最大,此后随浓度增加刺激作用逐渐减弱;酚红、血清中有雌激素样作用物质可以刺激雌激素依赖阳性的人乳腺癌细胞MCF-7生长,而MCF-7在无酚红去激素培养液中生长较为缓慢;受试物溶剂DMSO细胞增殖率低于无酚红去激素DMEM受试物溶剂,DMSO对人乳腺癌细胞MCF-7具有一定毒性;MCF-7细胞在去雌激素培养基中培养时间越长,细胞对雌二醇的刺激作用越敏感,培养时间达到7 d时,细胞对雌二醇的敏感性最高,MCF-7细胞与E2接触时间在48 h左右比较适宜,四种受试物雌激素效应从高到底排列为EE2>E2>E3>E1。结论 ATCC来源的MCF-7细胞对雌激素更加敏感;E2浓度为10-9 mol/L,以无酚红去激素DMEM为受试物溶剂;MCF-7细胞在去雌激素培养基中培养7 d,与E2接触48 h的条件下,MCF-7细胞对雌二醇的敏感性最高;四种受试物雌激素效应从高到低排列为EE2>E2>E3>E1。
Objective To study the effect of different test conditions on estrogen effect in MCF-7 proliferation assay. Methods MCF-7 cells from two different sources (ATCC and a tumor hospital) were used to study the effects of MCF-7 cells on the expression of 17β-estradiol (E2), ethynylestradiol Diol (EE2), estriol (E3), estrone (E1) reaction to optimize MCF-7 proliferation experimental conditions. Results The proliferation rate of MCF-7 cells from ATCC was higher than that of MCF-7 cells from tumor hospital under different concentrations of E2. With the increase of E2 concentration, the cell proliferation rate increased. When E2 concentration reached 10-9 mol / L , The cell proliferation rate was the highest, then stimulated gradually with the increase of concentration; Phenol red, estrogen-like substances in serum could stimulate the growth of estrogen-dependent human breast cancer cell line MCF-7, while MCF- The proliferation rate of DMSO in test substance solvent was lower than that in DMEM without phenol red extract. DMSO had certain toxicity to human breast cancer cell line MCF-7. MCF- The longer the culture time, the more sensitive the cells to the stimulation of estradiol. When the culture time reached 7 days, the cells had the highest sensitivity to estradiol and the contact time between MCF-7 cells and E2 was about 48 h Suitably, the estrogenic effects of the four test substances were ranked EE2> E2> E3> E1 from the highest to the lowest. CONCLUSIONS: MCF-7 cells derived from ATCC are more sensitive to estrogen. E2 concentration is 10-9 mol / L, and DMEM with no phenol red is used as the solvent. MCF-7 cells are cultured in estrogen medium d, the contact of E2 with E2 for 48 h, the sensitivity of MCF-7 cells to estradiol was the highest; the estrogenic effects of the four tested products were arranged as EE2> E2> E3> E1 from high to low.