小檗碱体外抗子宫内膜癌HEC-1A细胞的作用研究

Effects of berberine on endometrial carcinoma HEC-1A cell line in vitro

目的研究小檗碱体外对非激素依赖型人子宫内膜癌HEC-1A细胞增殖、迁移、周期、凋亡的影响。方法 MTT法检测小檗碱对人子宫内膜癌HEC-1A[中分化腺癌,雌激素受体(ER)弱阳性,孕激素受体(PR)阴性]细胞株的抑制作用;倒置显微镜观察细胞形态变化;将实验组分成50、25、12.5 mg/L高、中、低剂量组。Transwell法测定细胞体外迁移能力,流式细胞仪观察细胞周期分布,Annexin V-FITC/PI双染检测HEC-1A细胞凋亡。结果小檗碱可显著抑制HEC-1A细胞生长,在100、50、25、12.5、6.25 mg/L浓度范围内呈剂量依赖性(P<0.05)。小檗碱能明显改变细胞形态,小檗碱作用于子宫内膜癌HEC-1A细胞后,倒置显微镜下可见部分细胞体积改变,细胞内出现空泡,结构破坏,裂解成碎片,贴壁细胞减少,并散在排列。抑制子宫内膜癌HEC-1A细胞体外迁移的能力,高剂量组效果更佳,迁移细胞数明显减少。小檗碱低剂量组可以将HEC-1A细胞阻滞于G0/G1期。小檗碱作用24 h能提高HEC-1A细胞总凋亡率、早期凋亡率及晚期凋亡率,早期凋亡率更明显。结论小檗碱能够抑制非激素依赖型子宫内膜癌HEC-1A细胞株的增殖、迁移,并且促进HEC-1A细胞凋亡,低剂量小檗碱能将细胞阻滞在G0/G1期。

Objective To study the effects of berberine on proliferation,migration,cell cycle and apoptosis in hormone-independent endometrial carcinoma cell line HEC-1 A. Methods Berberine＇s inhibition of HEC-1 A cells proliferation was measured by using MTT colorimetry assay. Cell morphology was inspected under an inverted microscope. Investigated Berberine groups were divided into high-dose group（ 50 mg/L）,mid-dose group（ 25 mg/L）,and low-dose group（ 12. 5 mg/L）. Migration ability of cells in vitro was determined with Transwell. Cell cycle expressions were observed by using flow cytometry.Apoptosis of HEC-1 A cells was detected with Annexin V-FITC/PI double staining. Results Berberine may significantly inhibit the proliferation of HEC-1 A cells in a dose-dependent manner（ effective concentration of berberine ranged between 100 mg/L to 6. 25 mg/L）（ P 0. 05）. Berberine could markedly change cell morphology,which was visible under an inverted microscope,with vacuoles in cells,structural damage even cracking into pieces,decreased adherent cells,and scattered alignment. Berberine could significantly inhibit HEC-1 A cell migration especially in the high-dose group,on which amount the number of migrating cells decreased markedly. G0/G1 phase arrest was induced by low-dose group treatment after 24 hours. Berberine could accelerate total rate of apoptosis,early apoptosis rate and late apoptosis rate of HEC-1 A cells significantly; however,increase of early apoptosis rate was more apparrent.Conclusion Berberine could inhibit proliferation and migration of hormone-independent endometrial carcinoma cell line HEC-1 A,and improve the apoptosis rate of HEC-1 A. Low-dose berberine may induce the cell at G0/G1 phase.