雌激素对甲状腺乳头状癌细胞生长的影响及其机制

Effect of estrogen on the growth of papillary thyroid carcinoma cells and its mechanism

目的 观察雌激素对甲状腺乳头状癌BHP10-3细胞株生长的影响,并探讨其机制.方法 将生长良好的BHP10-3细胞根据随机数字表法分为实验组和对照组,实验组加入浓度为10-8mol/L的雌激素,对照组加入等量磷酸盐缓冲液（PBS）,噻唑蓝（MTT）测定BHP10-3细胞增殖,流式细胞仪测定BHP10-3细胞周期,Transwell侵袭迁移实验测定BHP10-3细胞侵袭迁移能力,聚合酶链反应（PCR）测定BHP10-3细胞人端粒酶反转录酶（hTERT）、细胞周期蛋白D1（Cyclin D1）、G蛋白耦联雌激素受体1（GPER1）、趋化因子受体1（CXCR1） mRNA表达量,Western blot测定BHP10-3细胞hTERT、Cyclin D1、GPER1、CXCR1蛋白表达量.结果 实验组BHP10-3细胞的增殖率[（18.34±1.67）%]明显高于对照组[（0.00±0.01）%,t=21.533,P=0.000].实验组BHP10-3细胞G1期细胞比例[（60.32±6.57）%]低于对照组[（71.21±8.44）%,t=4.153,P=0.004],实验组BHP10-3细胞S/G2期细胞比例[（37.48±3.12）%]高于对照组[（28.79±2.53）%,t=4.568,P=0.000].实验组侵袭细胞数[（42.31±4.35）个]和迁移细胞数[（46.51±4.57）个]均明显高于对照组[（17.35±2.54）个、（19.54±3.69）个;t=24.353,P=0.000;t=26.542,P=0.000）.实验组 BHP10-3细胞hTERT、Cyclin D1、GPER1、CXCR1 mRNA表达量（2.23±0.04、1.87±0.05、2.64±0.07、3.21±0.05） 均明显高于对照组（1.00±0.02、1.00±0.01、1.00±0.01、1.00±0.02;t=3.024,P=0.007;t=2.895,P=0.021;t=3.253,P=0.010;t=3.327,P=0.008）.实验组BHP10-3细胞hTERT、Cyclin D1、GPER1、CXCR1蛋白表达量（0.47±0.04、0.39±0.04、0.78±0.05、0.68±0.06） 均明显高于对照组（0.26±0.06、0.17±0.02、0.42±0.03、0.24±0.01;t=4.142,P=0.000;t=3.758,P=0.002;t=3.462,P=0.001;t=5.036,P=0.000）.结论 雌激素能够促进甲状腺乳头状癌BHP10-3细胞的增殖和侵袭转移,其机制可能与雌激素能够促进BHP10-3细胞hTERT、Cyclin D1、GPER1、CXCR1的表达量增加有关.

Objective To observe the effect of estrogen on the growth of papillary thyroid carcinoma cells, and to investigate its mechanism.Methods The well-grown BHP10-3 cells were divided into experimental group and control group according to random nubmer methods.The experimental group was added estradiol with concentration being 10-8 mol/L, the control group received the same amount of phosphate buffer （PBS） solution.The proliferation of BHP10-3 cells was detected by methyl thiazol tetrazolium （MTT） assay.The cell cycle of BHP10-3 cells was determined by flow cytometry.The invasion and migration ability of BHP10-3 cells were measured by Transwell invasion and migration assay.The human telomerase reverse transcriptase （hTERT） mRNA, Cyclin D1 mRNA, G protein coupled estrogen receptor 1 （GPER1） mRNA and chemokine receptor 1 （CXCR1） mRNA in BHP10-3 cells were determined by polymerase chain reaction （PCR）.The protein expressions of hTERT, Cyclin D1, GPER1 and CXCR1 in BHP10-3 cells were detected by Western blotting.Results The proliferation rate of BHP10-3 cells [（18.34±1.67）%] in the experimental group was significantly higher than that in the control group [（0.00±0.01）%, t=21.533, P=0.000].The percentage of G1 phase [（60.32±6.57）%] of BHP10-3 cells in the experimental group was lower than that of the control group [（71.21±8.44）%, t=4.153, P=0.004].The S/G2 phase of BHP10-3 cells in the experimental group was higher than that in the control group [（37.48±3.12）%, （28.79±2.53）%, t=4.568, P=0.000].The invasive cells （42.31±4.35） and migrating cells （46.51±4.57） in the experimental group were significantly higher than those in the control group （17.35±2.54, 19.54±3.69;t=24.353, P=0.000;t=26.542, P=0.000）.The expression levels of hTERT mRNA, Cyclin D1 mRNA, GPER1 mRNA and CXCR1 mRNA in BHP10-3 cells （2.23±0.04, 1.87±0.05, 2.64±0.07, 3.21±0.05） were significantly higher than those in the control group （1.00±0.02, 1.00±0.01, 1.00±0.01, 1.00±0.02;t=3.024, P=0.007;t=2.895, P=0.021;t=3.253, P=0.010;t=3.327, P=0.008）.The expression levels of hTERT protein, Cyclin D1 protein, GPER1 protein and CXCR1 protein in BHP10-3 cells （0.47±0.04, 0.39±0.04, 0.78±0.05, 0.68±0.06）were significantly higher than those in the control group （0.26±0.06, 0.17±0.02, 0.42±0.03, 0.24±0.01;t=4.142, P=0.000;t=3.758, P=0.002;t=3.462, P=0.001;t=5.036, P=0.000）.Conclusion Estrogen can promote the proliferation and invasion and metastasis of papillary thyroid carcinoma BHP10-3 cells, and its mechanism may be related to the estrogen can promote the hTERT, Cyclin D1, GPER1 and CXCR1 expression increased in BHP10-3 cells.