Anti-tumoral Effects And Mechanism Of Combination Of Icaritin And Cisplatin On Non-small Cell Lung Cancer

Background:As icaritin is a kind of traditional Chinese medicine ingredients, which has the function of protecting the cardiovascular system, promoting the growth and development of the skeleton, and enhancing the immunity. In addition, Further study found that icaritin has the anti-tumor effects of the different kinds of solid and non solid tumors.However,whether Icaritin combined with chemotherapy can enhance tumor sensitivity to chemotherapy is unclear, the specific mechanism of action is unclear.Objective:This study observed,in the cellular level, that cisplatin, Icaritin compares Icaritin combination with cisplatin,icaritin can be enhanced the anti-tumor effect of cisplatin and to study its possible mechanism.Methods:Icaritin, cisplatin and the combination group treated A549 cells.after different times, we observed A549 cells morphological changes. MTT assay was used to evaluate the inhibitory effects of Icaritin, cisplatin and the combination group treatment on the A549, BEAS-2B cells. The effect of Icaritin, cisplatin and the combination group on the clonal formation of A549 cells was detected by clonal and population experiments. Flow cytometry (FCM) assays and TUNEL assays were used to detect the cell cycle phase and cell apotosis in the A549 cell. The western blotting were applied to analyze the expressions level of AKT,p-AKT,Bcl-2,Bim,Caspase-9 proteins.Results:(1)Icaritin, cisplatin and combined group could make A549 cells morphological changes significantly, showing the typical characteristics of apoptosis cells.The cell morphological changes of the combined group were more obvious.(2)Icaritin could significantly inhibit the growth and proliferation in A549 cells,has a a time-dose-dependent relationship.(3) Cisplatin could significantly inhibit the growth and proliferation in A549 cells,has a a time-dose-dependent relationship.(4)Combination icaritin and cisplatin could significantly inhibit the growth and proliferation in A549 cells,has a time-dose-dependent relationship.The proliferation of A549 cells in the single-drug group was significantly higher than in the combination-group.According to Zhou’s formula calculated the combination index(CI),CI were less than 1.1,showing that the combination Icaritin and cisplatin could synergistically inhibit the proliferation of A549 cells.(5)Compared with cisplatin,Icaritin could lower inhibit proliferation of BEAS-2B cells, and the cytotoxicity of the cells was less. Combination Icaritin and cisplatin could not increase the cytotoxicity of BEAS-2B cells.(6)Icaritin and cisplatin could inhibit the colony formation of A549 cells. Compared with the single-drug group, the combination-group more significantly inhibited the colony formation of A549 cells.(7)Icaritin could make A549 cells arrest cell cycle at G2/M phase. Cisplatincould make A549 cells arrest cell cycle at G0/G1 and S phase. The combination-group could make A549 cells arrest cell cycle at S phase.(8)Icaritin,cisplatin and the combination-group could induce apoptosis of A549 cell. The combination-group could induce more significantly apoptosis of A549 cell, compared with the single-drug group.(9)The western blotting assay found combination Icaritin and cisplatin down-regulated expression level of AKT, p-AKT,Bcl-2,and decreased the rate of p-AKT/AKT,and up-regulated the expression level of Bim and Caspase-9 at protein level,but, compared with the single-drug group, the combination-group could regulate more significantly.Conclusions:Icaritin could inhibit non-small cell lung cancer(NSCLC) cells growth,induce NSCLC cells arrest and apoptosis,combination icaritin with cisplatin had a synergistic inhibitory effect on the NSCLC cells. This mechanism may be related to the apoptosis pathway mediated by PI3K/AKT signaling pathway.Application of the combination icaritin with cisplatin was treated lung cancer, reduced the toxicity of cisplatin, but also enhanced the anti-tumor effects,and did not appear toxicity of overlap. It is suggested that the combination icaritin with cisplatin has a certain feasibility and clinical application value in the treatment of lung cancer.