Inhibition Of Artesunate On Human Breast Cancer MCF7 And MDA-MB-231 Cells And Its Mechanisms

Objectives: To study the effect of Artesunate(ART)on human breast cancer cell line MCF7 and triple-negative breast cancer cell line MDA-MB-231 in cell proliferation,migration,invasion,cell cycle and DNA damage pathway proteins,The possible mechanism of artesunate inhibiting MCF7 and MDA-MB-231 cells in order to provide a theoretical basis for artesunate as a novel chemotherapeutic drug for breast cancer,especially triple-negative breast cancer.Methods:(1)Cells were treated with different concentrations of artesunate for 24 h,48h,and 72 h,respectively,MTT assay was used to detect the cell proliferation inhibition rates.(2)Crystal violet staining was used to detect the effects of artesunate on cell proliferation activity.(3)In order to reflect the impact on cell migration ability,cell scratch assay was used to detect the average scratch width under artesunate.(4)The number of invasive cells of MDA-MB-231 cells treated with artesunate was determined by Transwell chamber assay.(5)Flow cytometry was used to detect the effects of artesunate on the cell cycle.(6)Western blot was used to detect the effects of artesunate on the DNA damage response pathway proteins,such as?H2AX(S139),phospho-ATM(S1981),phospho-CHK2(T68)and phospho-CDC25C(S216).Results:(1)MTT results showed that with the increase of artesunate concentrations the proliferation inhibition rates of MCF7 cells and MDA-MB-231 cells increased(P<0.05)in a concentration-dependent manner.(2)The results of crystal violet staining showed that the cell density of MCF7 and MDA-MB-231 decreased gradually with the increase of artesunate concentration,and the OD values of crystal violet dissolved gradually decreased(P<0.05).(3)Cell scratch test results showed that the average scratch width of MCF7 and MDA-MB-231 cells was significantly increased under the action of artesunate(P<0.05),and the cell migration distance was reduced.(4)The results of Transwell showed that the number of invasive cells in MDA-MB-231 cells was significantly decreased with the increase of artesunate concentration(P<0.05).(5)The results of flow cytometry showed that with the increase of artesunate concentration,the proportion of cells in G0/G1 phase of MCF7 and MDA-MB-231 decreased gradually(P<0.05),and the proportion of cells in G2/M phase increased gradually(P<0.05).(6)Western blot results showed that the protein levels of ?H2AX(S139),phospho-ATM(S1981),phospho-CHK2(T68)and phospho-CDC25C(S216)in MCF7 and MDA-MB-231 experimental groups were significantly increased(P<0.05).Conclusions:(1)Artesunate can inhibit the proliferation of MCF7 and MDA-MB-231 cells in a concentration-dependent manner.(2)Artesunate can inhibit the in-vitro migration of MCF7 cells and MDA-MB-231 cells.(3)Artesunate can inhibit the invasiveness of MDA-MB-231 cells in vitro.(4)Artesunate can induce G2/M arrest in MCF7 and MDA-MB-231 cells.(5)The possible inhibition mechanism of artesunate on MCF7 and MDA-MB-231 cells was that Artesunate induced G2/M cell cycle arrest and DNA damage response by activation of proteins such as ?H2AX(S139),phospho-ATM(S1981),phospho-CHK2(T68)and phospho-CDC25 C,resulting in the inhibition on MCF7 and MDA-MB-231 cells.