鞣花酸对BRCA1沉默的三阴性乳腺癌MDA-MB-231细胞生物学行为的影响及其作用机制

Effect of ellagic acid on the biological behaviors of BRCA1 silenced triple negative breast cancer cell line MDA-MB-231 and its possible machanism

目的探讨鞣花酸(ellagic acid,EA)对乳腺癌易感基因1(breast cancer susceptibility gene 1,BRCA1)沉默的三阴性乳腺癌MDA-MB-231细胞生物学行为的影响及可能的作用机制。方法采用不同浓度EA(0μg/mL、1μg/mL、5μg/mL、10μg/mL和20μg/mL)分别作用MDA-MB-231细胞24 h、48 h和72 h后,用MTT法检测细胞增殖能力,并分别计算其IC50,筛选最佳药物浓度。将BRCA1 siRNA转染MDA-MB-231细胞,以终浓度为5μg/mL的EA作用0 h、24 h、48 h、72 h、96 h,采用MTT法检测细胞增殖能力,划痕实验检测细胞迁移能力,Transwell小室检测细胞迁移和侵袭能力,RT-qPCR和Western blot检测PARP1 mRNA和蛋白的表达情况。结果不同浓度EA作用MDA-MB-231细胞24 h、48 h和72 h后均能抑制细胞增殖,IC50分别为13.739μg/mL、10.645μg/mL、5.344μg/mL。EA作用48 h、72 h和96 h后,siRNA+EA组的OD值均明显小于对照组、EA组和阴性对照组(P<0.001)。siRNA+EA组细胞的迁移率及穿膜细胞数均较其他3组低(P<0.001),且PARP1 mRNA和蛋白表达量亦均较低(P<0.001)。结论EA可抑制BRCA1沉默的三阴性乳腺癌MDA-MB-231细胞的增殖、迁移和侵袭,其作用机制可能与抑制DNA修复通路关键因子PARP1表达有关。

Objective To investigate the effects of ellagic acid(EA)on the biological behaviors of breast cancer susceptibility gene 1(BRCA1)silenced in triple negative breast cancer cell line MDA-MB-231 and its possible mechanism.Methods Different concentrations of EA(0μg/mL,1μg/mL,5μg/mL,10μg/mL and 20μg/mL)were used to treat MDA-MB-231 cells for 24 h,48 h and 72 h,respectively.The inhibition rate of proliferation were assessed by MTT assay and the IC50 was calculated to select the optimal drug concentration.After transfecting BRCA1 siRNA into MDA-MB-231 cells,EA was applied at a final concentration of 5μg/mL for 0 h,24 h,48 h,72 h,96 h,and cell proliferation was detected by MTT assay.Scratch assay was used to detect cell migration,Transwell chamber was used to detect cell migration and invasion,RT-qPCR and Western blot were used to detect the expression of PARP1 mRNA and protein.Results Different concentrations of EA had significant inhibitory effect on the proliferation of MDA-MB-231 cells at 24 h,48 h and 72 h,and IC50 were 13.739μg/mL,10.645μg/mL,5.344μg/mL,respectively.The OD value in siRNA+EA group was significantly lower than that of control group,EA group and negative control group(P<0.001). The cell migration rate and the number of transmembrane cells in siRNA+EA group were lower than those of the other three groups(P<0.001),and the expression of PARP1 mRNA and protein in siRNA+EA group were also significantly lower than those of the other three groups(P<0.001). Conclusions EA can inhibit the proliferation,migration and invasion of BRCA1 silenced MDA-MB-231 cells in vitro,and its mechanism may be related to related DNA repair pathway disorders.