凌晓斐, 胥可, 孙亚昕, 邵华, 王翠娟. 镉通过类雌激素样作用促进人乳腺癌MCF-7细胞增殖[J]. 环境与职业医学, 2018, 35(4): 356-360. DOI: 10.13213/j.cnki.jeom.2018.17604
引用本文: 凌晓斐, 胥可, 孙亚昕, 邵华, 王翠娟. 镉通过类雌激素样作用促进人乳腺癌MCF-7细胞增殖[J]. 环境与职业医学, 2018, 35(4): 356-360. DOI: 10.13213/j.cnki.jeom.2018.17604
LING Xiao-fei, XU Ke, SUN Ya-xin, SHAO Hua, WANG Cui-juan. Estrogen-like effect of cadmium on promoting proliferation of human breast cancer MCF-7 cells[J]. Journal of Environmental and Occupational Medicine, 2018, 35(4): 356-360. DOI: 10.13213/j.cnki.jeom.2018.17604
Citation: LING Xiao-fei, XU Ke, SUN Ya-xin, SHAO Hua, WANG Cui-juan. Estrogen-like effect of cadmium on promoting proliferation of human breast cancer MCF-7 cells[J]. Journal of Environmental and Occupational Medicine, 2018, 35(4): 356-360. DOI: 10.13213/j.cnki.jeom.2018.17604

镉通过类雌激素样作用促进人乳腺癌MCF-7细胞增殖

Estrogen-like effect of cadmium on promoting proliferation of human breast cancer MCF-7 cells

  • 摘要: 目的 研究镉对人乳腺癌MCF-7细胞增殖及雌激素受体(estrogen receptor,ER)蛋白表达水平的影响及其可能机制。

    方法 用1×10-6、1×10-7、1×10-8、1×10-9、1×10-10、1×10-11 mol/L的17β-雌二醇(后称雌激素)培养不同来源的A、B两株MCF-7细胞4 d,应用CCK8法筛选敏感细胞株并确定雌激素促细胞增殖作用最强的浓度。用1×10-6、1×10-7、1×10-8、1×10-9、10-10、1×10-11 mol/L的镉溶液染毒敏感细胞株,确定镉促进细胞增殖的最大浓度。设置阴性对照组(去雌激素培养液)、实验组(1×10-8 mol/L镉)、阳性对照组(1×10-9 mol/L雌激素),通过克隆形成实验检测镉对MCF-7细胞集落形成能力的影响。利用ER抑制剂(ICI182780)初步探讨镉致细胞增殖的可能机制,增设实验抑制剂组(1×10-8 mol/L镉+1×10-7 mol/L ER抑制剂)、阳性抑制剂组(1×10-9 mol/L雌激素+1×10-7 mol/L ER抑制剂),通过CCK8法、流式细胞术和Western blot分别检测各组的细胞增殖率、细胞周期S期比例和ER表达水平。

    结果 实验所用B株MCF-7细胞为雌激素敏感细胞株,且当雌激素浓度为1×10-9 mol/L时,对MCF-7细胞的促进增殖作用(203.55±36.65)%最大(P<0.001)。与阴性对照组(100%)相比,1×10-8~1×10-10 mol/L镉溶液可促进MCF-7细胞增殖(163.78±31.90)%~(176.88±10.06)%(P<0.001)。1×10-8 mol/L镉可促进细胞集落形成(P<0.05),增加细胞S期比例(P<0.001),提高细胞ER蛋白表达水平(P<0.001)。与实验组相比,加入ER抑制剂能够抑制镉对MCF-7细胞的促增殖作用由(135.17±23.96)%降至(107.66±7.64)%(P<0.01),抑制镉诱导的细胞S期比例增加由(24.17±0.53)%降至(12.36±0.43)%(P<0.001),拮抗镉诱导的ER表达增加由(56.19±3.67)%降至(38.84±1.04)%(P<0.05)。

    结论 镉可以促进人乳腺癌细胞MCF-7增殖和ER表达,这种作用可被ER抑制剂所抑制,提示镉可能具有雌激素样作用。

     

    Abstract: Objective To investigate the effect and potential mechanism of cadmium (Cd) on the proliferation of human breast cancer MCF-7 cells and the expression of estrogen receptor (ER).

    Methods A and B strains of MCF-7 cells were treated with 17β-estradiol (hereinafter referred to as estrogen) at 1×10-6, 1×10-7, 1×10-8, 1×10-9, 1×10-10, and 1×10-11 mol/L, respectively, for 4 d. CCK8 assay was employed to detect a susceptible MCF-7 cell strain and to identify the concentration of estrogen promoting maximum cell proliferation. Then, the susceptible MCF-7 cell strain was treated with 1×10-6, 1×10-7, 1×10-8, 1×10-9, 1×10-10, and 1×10-11 mol/L cadmium solution, respectively, to identify the concentration of cadmium promoting maximum cell proliferation. We set a negative control group (medium without estrogen), an experimental group (1×10-8 mol/L cadmium), and a positive control group (1×10-9 mol/L estrogen) to evaluate colony formation ability by clonogenicity assay. Additionally, we set an experimental inhibitor group1×10-8 mol/L Cd+1×10-7 mol/L ER antagonist (ICI182780) and a positive inhibitor group (1×10-9 mol/L estrogen+1×10-7 mol/L ER antagonist) to detect cell proliferation, the ratio of S phase cells, and the expression level of ER by CCK8, flow cytometry, and Western blot, respectively.

    Results The MCF-7 cells of strain B were estrogen sensitive cells, and the 1×10-9 mol/L estrogen treatment induced the maximum cell proliferation(203.55±36.65)%. Compared with the negative control group (100%), 1×10-8-1×10-10 mol/L cadmium significantly promoted proliferation of MCF-7 cells(163.78±31.90)%-(176.88±10.06)% (P < 0.001). Besides, 1×10-8 mol/L cadmium treatment promoted colony formation of MCF-7 cells (P < 0.05), increased the ratio of S phase cells (P < 0.001), and elevated ER protein expression level (P < 0.001). Compare with the experimental group, ER antagonist suppressed the effect of cadmium on proliferation of MCF-7 cellsfrom (135.17±23.96)% to (107.66±7.64)% (P < 0.01), reduced the ratio of S phase cellsfrom (24.17±0.53)% to (12.36±0.43)% (P < 0.001), and down-regulated ER protein expression levelfrom (56.19±3.67)% to (38.84±1.04)% (P < 0.05).

    Conclusion Cadmium can promote proliferation and ER expression of breast cancer MCF-7 cells, and the effect can be inhibited by ER antagonist, indicating estrogen-like effect of cadmium.

     

/

返回文章
返回