低剂量双酚A及邻苯二甲酸二乙基己酯对PC3细胞增殖的影响

Effects of low-dose Bisphenol A and DiethylheXylPhthalate on PC3 cells proliferation

目的:探讨低剂量环境内分泌干扰物双酚A(Bisphenol A,BPA)和邻苯二甲酸二乙基己酯(Di-2-ethylhexy-lPhthalate,DEHP)对PC3细胞增殖的影响。方法:PC3细胞用含10%胎牛血清的F12培养基培养,将处于对数生长期的细胞制成104/ml的细胞悬液,按100μl/孔容积加入96孔板,24 h后分别加入BPA、DEHP、雌二醇(Estradiol,E2)和双氢睾酮(Dihydrotestosterone,DHT)10μl/孔,终浓度为0、0.0001、0.001、0.01、0.1、1、10、100、1 000、10 000、100 000 nM,每个剂量设3个复孔,重复2次,24 h后加入CCK8 10μl/孔,37℃孵育4 h后450 nm处检测吸光度;然后在培养液中加入终浓度为0.1μM/ml的他莫昔芬(Tamoxifen,Tam)拮抗雌激素受体(Estradiol Receptor,ER)的作用,用能明显促进细胞增殖的0.01、0.1、1、10、100 nM剂量,再次对PC3细胞进行染毒,同样方法检测4种物质作用PC3后的吸光度。结果:10 nM和100 nM的BPA,0.01 nM、0.1 nM和10nM的E2,0.01、0.1、1、10、100、1000、10 000、100 000 nM的DEHP均有促进PC3细胞增殖的作用,而DHT却无这种影响,用TAM封闭ER功能后,0.1、1、10、100nM的BPA、DEHP、E2、DHT均有促进PC3增殖的效果。结论:低剂量环境内分泌干扰物BPA和DEHP可促进PC3细胞增殖,作用机制与E2相似;而当ER被封闭后,DHT却呈现出促细胞增殖作用。

Objective： To explore the effects of low doses of environmental endocrine disrupting chemicals bisphenol A （BPA） and DiethylhexylPhthalate （DEHP） on PC3 cells proliferation. Methods： PC3 cells were cultured with F12 medium include 10% fetal bovine serum at 37℃, 5% CO2, Cell suspension was added to 96 well plate, and each well had 1000 cells for 24h, after cells were exposed to 0, 0.0001, 0.001, 0.01, 0. 1, 1, 10, 100, 1000, 10000, 100000nM concentration of BPA, DEHP, Estradio （E2） and （Dihydrote- stosterone, DHT） , each dose for three well and repeat 2 times, respectively for 24h, cell proliferation was detected with CCK-8 assay. PC3 cells were cultured with F12 medium include 0. 1μM Tamoxifen （TAM）, after cells were ex posed to 0, 0. 01, 0. 1, 1, 10, 100, 1000nM concentration of BPA, DEHP, E2 and DHT, Absorbance was detected with CCK- 8 assay. Results： 10nM and 100nM of the BPA, 0.01nM, 0. lnM and 10nM orE2, 0.01, 0. 1, 1, 10, 100, 1000, 10000, 100000 nM of DEHP have the duty to promote the role of PC3 cell proliferation, while no such effects DHT, ER closed by TAM feature, 0. 1, 1, 10, 100 nM ofBPA, DEHP, E2 and DHT has the effect of promoting the proliferation of PC3. Conclusion： The low dose of environmental endocrine disruptors BPA and DEHP can promote PC3 cell proliferation, similar to the role of the mechanism and E2 ; and when the ER was closed, DHT show the role of promoting cell proliferation.