The Effect Of S100A4Monoclonal Antibody Combined With Paclitaxel On The Proliferation And Apoptosis Of Hela Cells

Background and ObjectiveCervical cancer is the most common gynecological malignancy among women.Treatment strategies including surgery and radiotherapy have a significant impact on quality of patients’life.So chemotherapy is getting more and more attention.Paclitaxel(PTX)is one of the most active chemotherapeutic drugs used to treat cervical cancer,which can prevent the proliferation and division of tumor cells,prevent mitosis in the process of cell division at the G2/M phase,result the apoptosis of cells.Because of the side effects including agranulocytosis induced by myelosuppression,allergic reactions,hepatic toxicity and neurotoxicity could still not be avoided,so new ways need to be found to minimise its dosages and side effects.S100A4(S100calcium binding protein A4)is highly expresed in cervical carcinoma and closely related to tumor cell invasion and metastasis and clinical stages.Currently targeted cancer therapies are drugs or other substances that block the growth and spread of cancer by interfering with specific molecules involved in tumor growth and progression.Monoclonal antibody is one of the most studied new drugs in particular.Through tetramethyl azo blue(MTT)colorimetric method and flow cytometric detection(FCM)method we explored different concentrations of PTX,S100A4monoclonal antibodies as well as S100A4monoclonal antibody combined with PTX on the growth of Hela cells in vitro,looking forward to providing effective theoretical basis for the treatment of cervical cancer.Materials and MethodsGeneral ways was made for Hela cells recovery in vitro successfully.Then we observed cell growth and morphological changes.Groups:1.paclitaxel monotherapy groups:drug concentrations were lOug/ml,20ug/ml,40ug/ml,80ug/ml,and160ug/ml.2.groups of S100A4monoclonal antibody:antibody concentrations were lug/ml,5ug/ml:10ug/mL,20ug/ml.3.the combined group:S100A4monoclonal antibody concentration of10ug/mL combined with paclitaxel individual concentrations(10μg/ml,20μ g/ml,40μg/ml,80μg/ml,160μg/ml).4.The blank group was only given the same concentration of1640medium.5.The control groups was given the same concentration of1640medium and Hela cells additionally without any interference factors.Methods:1.According to the drug intervention we cultured the cells respectively for24h,48h,72h cell and used MTT method to detect cell inhibition rate.2.Flow cytometry was used to detect cell apoptosis and cell cycle distribution after the drug intervention and24hours’cell culture.SPSS17.0Statistical analysis software was used to make a reasonable statistical analysis of the experimental data,a=0.05was considered as the detection standard.Results1Using an inverted microscope we observed the size and shape of Hela cells.2By MTT colorimetric method,the inhibition on the growth of Hela cells was increased gradually with time-and dose-dependent in the range of10ug/mL,20ug/ml,40ug/ml,80ug/ml,160ug/ml PTX concentration.There were statistically significant differences between any of the groups(p<0.05).3By MTT colorimetric method,the inhibition on the growth of Hela cells was increased gradually with time-and dose-dependent in the range of lug/ml,5ug/ml,lOug/ml,20ug/ml S100A4monoclonal antibodies concentration.There were statistically significant differences between any of the groups(p<0.05).4S100A4antibody lOug/ml combined with paclitaxel(10p.g/ml,20Mu g/ml,40,Mu g/ml,80Mu g/ml,160μg/ml)showed higher inhibition rate of Hela cells than that of the paclitaxel monotherapy.There were statistically significant differences between any of the groups(p<0.05).5Apoptosis rate of Hela cells detected by Flow cytometric after drug intervention for24hours was(0.95±0.11)%for control group,(31.85±0.52)%for paclitaxel monotherapy group,(6.09±0.24)%for S100A4monoclonal antibody group,(35.64±0.83)%for the paclitaxel combined with S100A4monoclonal antibody group respectively,there were statistically significant differences between any of the groups(p<0.05).Analysis of cell cycle distribution comparison for all groups:The proportion of G0/G1phase and S phase in paclitaxel monotherapy group was decreased than that in the control group,with G2/M phase increased than that in the control group.The proportion of G0/G1phase in S100A4monoclonal antibody group was increased than that in the control group,with G2/M phase and S phase decreased than that in the control group.The proportion of G0/G1phase and G2/M phase in the paclitaxel combined with S100A4monoclonal antibody group was increased than that in the paclitaxel monotherapy group,with S phase decreased than that in the paclitaxel monotherapy group.Conclusion1Paclitaxel had the significant inhibitory effect on the growth and proliferation of Hela cells with time-and dose-dependent.2S100A4monoclonal antibody had the significant inhibitory effect on the growth and proliferation of Hela cells with time-and dose-dependent.3Paclitaxel combined with S100A4monoclonal antibody could improve the sensitivity of Hela cells to paclitaxel,which had synergistic antitumor effect.