The Inhibitory Effect Of Chlorpromazine On Human Triple Negative Breast Cancer Cells

Background:High heterogeneity is one of features of triple-negative breast cancer(TNBC).Due to the absence of estrogen receptor(ER),progesterone receptor(PR)and human epidermal growth factor receptor 2(HER2),there is still a lack of effective treatment for triple negative breast cancer besides conventional surgery and chemotherapy.Therefore,to explore new methods for triple negative breast cancer has been a hotspot.In recent years,many study suggested that antipsychotics chlorpromazine had inhibitory effect on a variety of malignant tumors,but yet in TNBC for further study.Based on above,we deduced that if chlorpromazine had an inhibitory effect on TNBC.Objective: To investigate if the chlorpromazine can inhibit human TNBC cancer cells and its possible mechanism.Methods: The inhibitory effect of chlorpromazine on the proliferation of human TNBC cells was determined by CCK-8,and the IC50 was calculated.The inhibitory effect of chlorpromazine on migration was determined by scratch test.The effect of chlorpromazine on apoptosis and cycle was detected by flow cytometry.The expression of Bcl2 and Bax were detected by q RT-PCR and WB when chlorpromazine act on TNBC cells.At the same time,the expression of oncogene CIP2 A and c-Myc were detected by q RT-PCR and WB.Results: The results of CCK-8 demonstrated that different concentration of chlorpromazine had a significant inhibitory effect on cell proliferation in human TNBC cell lines(MDA-MB-231,SUM159PT).The IC50 of chlorpromazine against MDAMB-231 and SUM159 PT cells were(20.78±2.07)μM and(16.74±0.93)μM,respectively.The scratch test results demonstrated that the wound healing rate of treatment group was lower than control group.The results of Flow cytometry revealed that MDA-MB-231 and SUM159 PT had cycle arrest and apoptosis.Meanwhile,q RTPCR and WB indicated that the expressions of m RNA and protein of Bax increased,while the the expressions of m RNA and protein of Bcl-2 decreased when chlorpromazine act on MDA-MB-231 and SUM159 PT.At the same time,the m RNA and protein of CIP2 A and c-Myc in TNBC cells were decreased by chlorpromazine.Conclusions: Chlorpromazine inhibit the proliferation and migration of MDAMB-231 and SUM159 PT,and induce their cycle arrest and apoptosis.The mechanism of apoptosis induction is related to the down-regulation of Bcl-2 expression and upregulation of Bax expression.In addition,chlorpromazine can also down-regulate the expression of oncogenes CIP2 A and c-myc in MDA-MB-231 and SUM159 PT in TNBC cells,which may exert its inhibitory effect through PP2A/CIP2A/ c-Myc.This paper will elaborate the inhibitory effect of chlorpromazine on TNBC in three parts:Part1:Chlorpromazine inhibits the proliferation and migration of human TNBC cells.Part2: Chlorpromazine induced apoptosis and cycle arrest in TNBC cells,and the mechanism of apoptosis was related to the down-regulation of Bcl-2 and up-regulation of Bax.Part3:chlorpromazine mediated its inhibitory effect on TNBC cells by downregulating the oncogene CIP2 A and c-Myc.