The Effect Of Huaier Extract On The Proliferation,Apoptosis And Tamoxifen Resistance In Breast Cancer Cells

Objective Breast cancer has seriously threaten women's health and the incidence rate is still on the rise.Patients with ER positive breast cancer will eventually resistant to tamoxifen treatment after a long period of antiestrogen intervention.Upon patients having developed TAM resistance,they will benefit little from endocrine therapy.TAM resistance adversely influence the prognoses of ER positive breast cancer,thus it has become a crucial clinic problem.However,Chinese traditional medicine Huaier has been found capable of multiple anti-tumor effect,which may provide a possibility to overcome tamoxifen resistance.This study intend to investigate the effect of Huaier extract on the proliferation and apoptosis of human breast cancer cell lines MCF-7?LCC2,as well as the influence on tamoxifen resistance.Methods Human breast cancer cell lines MCF-7 and LCC2 was cultured in vitro and treated by different concentrations of Huaier extract.MTT assay was used to detect the effect of Huaier on the proliferation of MCF-7 and LCC2 cells.It was also used to examine the combinatorial effect of Huaier and tamoxifen on TAM resistant cells.Flow cytometry was used to detect the apoptosis status of MCF-7 and LCC2 cells after 48 h Huaier treatment.Western blot method was used to measure the expression of tamoxifen resistance related protein in LCC2 cells,such as ER?,AIB1 and Survivin,after 48 h gradient of Huaier treatment.The significant differences between multiple groups were statistically analysised by One-way ANOVA,and the differences between two groups were analysised by t-test.Results 1.MTT assay showed that the proliferation of MCF-7 and LCC2were significantly inhibited by Huaier treatment,compared with control group,the differences have statistic significance.The IC₅₀0 of two cells at 24 h,48 h,72 h were?9.58±0.52?6.43±0.20?4.25±0.13 mg/mL?vs?18.16±0.16?13.10±0.97?8.66±0.18 mg/mL?,respectively,which implies that MCF-7 cells were more sensitive to Huaier than LCC2 cells.2.The results of flow cytometry indicated that Huaier extract was able to induce apoptosis of MCF-7 and LCC2 cells.Moreover,the number of the early apoptotic cells increased with the drug concentration.The number of early apoptotic cells of MCF-7,after concentrations of 0?2?4?8 mg/mL Huaier treatment,were?2.03±0.61?%??6.27±1.16?%??12.53±0.57?%??14.27±1.10?%,respectively,one-way analysis of variance,F=118.42,P<0.001.Similarly,the early apoptotic cells number of LCC2 at the different concentrations of Huaier treatment were?3.33±0.64?%??7.17±1.25?%??10.27±2.02?%??12.77±0.21?%,respectively,one-way analysis of variance,F=33.59,P<0.001.3.After gradient of Huaier intervention?0?2?4?8?16 mg/mL?,the expression of ER??AIB1 and Survivin in LCC2 were detected by western blot.As the drug concentration increasing,the expresson of ER?decreased gradually.Before huaier treatment,compared with MCF-7,AIB1 and Survivin highly expressed in LCC2 cells,while when the LCC2 cells were treated by huaier,the expression of AIB1 and Survivin down-regulated evidently.4.Finally,MTT assay verified that both the concentration of 2 and 4 mg/mL Huaier could enhance the sensitivity of LCC2 to tamoxifen.Furthermore,the higher drug concentration was more potent to enhance the inhibitory effect of tamoxifen on the growth of TAM-resistant cells.Conclusion Huaier extract can inhibit the proliferation of MCF-7 and LCC2 cell lines and induce its apoptosis in a concentration dependent manner.Huaier can also decrease the expression of ER?,AIB1 and survivin in tamoxifen resistant LCC2 cells,and enhance the sensitivity of LCC2 to tamoxifen treatment.