In Vitro Study Of NK Cells Combined With Lobaplatin On Human Breast Cancer MDA-MB-231 Cells

Objective:To investigate the effect of lobaplatin combined with NK immune cells on human breast cancer MDA-MB-231 cells by in vitro experiment,and to observe whether the killing effect of lobaplatin combined with NK cells on MDA-MB-231 cells is better than that of lobaplatin alone or NK cells alone,and to investigate the Caspase-3 when lobaplatin combined with NK cells kill MDA-MB-231 cells The expression of 3 and IL-23 and their possible roles provide theoretical support for the application of lobaplatin combined with NK cells in the clinical treatment of breast cancer.Methods:Human breast cancer cell line MDA-MB-231 frozen in the laboratory of Cancer Biotherapy Research Center of Inner Mongolia Cancer Hospital was cultured in vitro after resuscitation.Lobaplatin experimental group,NK experimental group,combined experimental group,NK control group,NK lobaplatin experimental group,target cell control group,blank group were set up.The optimal time and target ratio of NK cells were measured by CCK-8 method,and the proliferation inhibition rate of NK cells and lobaplatin alone or in combination on human breast cancer MDA-MB-231 cells was measured and calculated.Flow cytometry was used to detect and analyze the apoptosis rate of human breast cancer MDA-MB-231 cells in NK cell alone group,lobaplatin alone group,combined group and target cell control group.Western blot was used to detect the expression of interleukin-23 and Caspase-3 proteins in the NK cell alone group,lobaplatin alone group,combined group and target cell control groupResults:CCK-8 cell proliferation assay results:(1)In the same treatment time,the inhibitory rate of lobaplatin on human breast cancer MDA-MB-231 cell proliferation increased with lobaplatin concentration increasing(P<0.05).At the same concentration of lobaplatin,the inhibition rate increased with time(P<0.05);(2)The inhibition rate of NK cells on human breast cancer MDA-MB-231 cells decreased with the extension of time from 2 hours to 6 hours,and the inhibition rate was the highest at 2 hours(P<0.05).When the effective target ratio was 10:1,20:1 and 40:1,the inhibition rate of cancer cell proliferation increased with the increase of the effective target ratio(P<0.05),From 40:1 to 80:1,the inhibition rate of cancer cell proliferation decreased with the increase of effective target ratio(P<0.05);(3)The proliferation inhibition rate of NK cells combined with different concentrations of lobaplatin on human breast cancer MDA-MB-231 cells was higher than that of NK cells alone at the optimal acting time(P<0.05).The proliferation inhibition rate of MDA-MB-231 cells was higher than that of lobaplatin alone(P<0.05);(4)NK cells combined with lobaplatin at the lowest concentration(5ug/ml)had a higher proliferation inhibition rate on MDA-MB-231 cells than lobaplatin alone at high concentration(20 and 40ug/ml)(P<0.05).At the same time,lobaplatin combined with optimal target NK cells had additive effect on the proliferation inhibition of MDA-MB-231 cells.Flow cytometry results:(1)the apoptosis rate of MDA-MB-231 cells increased with the increase of lobaplatin concentration(P<0.05),and was higher than that of the control group(P<0.05).The average apoptosis rate of MDA-MB-231 cells treated with NK cells alone was higher than that of control group(P<0.05).The apoptosis rate of MDA-MB-231 cells after NK cells combined with different concentrations of lobaplatin increased with the increase of lobaplatin concentration(P<0.05),which was higher than that of lobaplatin alone or NK cells alone(P<0.05).The apoptosis rate of MDA-MB-231 cells induced by lobaplatin combined with NK cells at low concentration was higher than that induced by high concentration lobaplatin alone(P<0.05).Western blot results:IL-23 and Caspase-3 proteins were detected in lobaplatin group,NK group,combination group and control group.Caspase-3 protein expression:lobaplatin group and NK group were higher than control group(P<0.05),combined group was higher than corresponding concentration lobaplatin group(P<0.05),combined group was higher than NK group(P<0.05).IL-23 protein expression:lobaplatin group and NK group were higher than control group(P<0.05),combined group was higher than lobaplatin group(P<0.05).The combined histone expression of lobaplatin at 20ug/ml and 10ug/ml was higher than that of NK group(P<0.05),and the difference was statistically significant.The combined group with lobaplatin concentration of 5ug/ml had no statistical significance compared with the NK group.Conclusions:In vitro experiments,both lobaplatin and NK cells can significantly inhibit the proliferation of human breast cancer MDA-MB-231 cells,and both can cause the apoptosis of human breast cancer MDA-MB-231 cells The proliferation inhibition rate and apoptosis rate of human breast cancer MDA-MB-231 cells in combination with lobaplatin were significantly higher than those in combination with lobaplatin and NK cells alone,and the proliferation inhibition rate and apoptosis rate of human breast cancer MDA-MB-231 cells in combination with lobaplatin at low concentration and optimal target ratio were significantly higher than those in combination with lobaplatin at high concentrationThe expression levels of IL-23 and caspase-3,the key modulator of apoptosis,may be related to the killing degree of lobaplatin NK cells and cancer cells after lobaplatin NK cells and their combined action on human breast cancer MDA-MB-231 cells.Reducing the dosage of lobaplatin combined with NK cells to achieve the same or better therapeutic effect while reducing the toxic and side effects of lobaplatin may be a better choice for the treatment of triple negative breast cancer.In addition,inhibiting the expression of inflammatory factor IL-23 and caspase-3,a key regulator of apoptosis,may be beneficial for the treatment of triple negative breast cancer.