Studies On The Effect Of Dehydrocarvedine Combined With Cisplatin On Lung Cancer Cells

Objective: Dehydrocarvetin,the main active compound extracted from berberine,is involved in the regulation of cell behavior of many kinds of cells,but the effect of dehydrocarvetin on lung cancer cells has not been studied at home and abroad.The aim of this study was to investigate the effects of single and combined use of dehydrocarvetin and cisplatin on the proliferation and apoptosis of lung cancer cells.Methods: In vitro culture of human lung adenocarcinoma cell line A549 to logarithmic growth phase.CCK8 was used to detect the dehydrocarvedin group（0g/L、0.001g/L、0.002g/L、0.005g/L、0.01g/L）,cisplatin and dehydrocarvedin combined with cisplatin at 24h、48h、72h later,to observe the inhibitory effect on proliferation of human lung adenocarcinoma cell line A549;After 24 hours of single drug dehydrocarvetin,cisplatin and dehydrocarvetin combined with cisplatin,The apoptosis of human lung adenocarcinoma A549 cell line was measured by Annexin V/PI double staining flow cytometry.RT-PCR was used to detect the expressions of caspase-3 and Survivin m RNA in cells treated with single drug dehydrocarvetin,cisplatin,and dehydrocarvetin combined with cisplatin for 24 h.Western blot was used to detect the protein expression of caspase-3,Survivin in cells treated with single drug dehydrocarvetin,cisplatin,and dehydrocarvetin combined with cisplatin for 24 h.Results: The analysis of the inhibitory rate of CCK8 on human lung adenocarcinoma A549 cells showed that with the increase of Dehydrocarvetin concentration and the prolongation of the action time,the inhibitory effect of CCK8 on human lung adenocarcinoma A549 cells showed a time-dose effect relationship.There was significant difference between the single drug group and the corresponding combination group.The inhibitory effect of the combination group was significantly higher than that of the respective single drug group.Flow cytometry was used to detect the apoptosis of human lung adenocarcinoma A549 cells by using Dehydrocarvetin 0.2g/L alone,cisplatin 0.006g/L alone and Dehydrocarvetin combined with cisplatin,and the combined treatment group increased the apoptosis of human lung adenocarcinoma A549 cells in a synergistic way.Reverse transcription-polymerase chain reaction（RT-PCR）results showed that compared with the negative control group,dehydrocarvetin 0.2g/L group,cisplatin0.006g/L group,and dehydrocarvetin combined with cisplatin group could up-regulate the expression of caspase-3 m RNA and down-regulate the expression of Survivin m RNA.There were significant differences between the combined group and other groups.Western blot analysis showed that compared with the negative control group,dehydrocarvetin 0.2g/L group,cisplatin0.006g/L group and dehydrocarvetin combined with cisplatin group could up-regulate the expression of caspase-3 protein and down-regulate the expression of Survivin protein,There are significant differences between the combined group and other groups in the down-regulated expression of Survivin protein.Conclusions: This study demonstrated that Dehydrocarvetin combined with cisplatin could significantly inhibit the proliferation and promote apoptosis of human lung cancer A549 cells.Its synergistic effect may be related to the down-regulation of survivin and up-regulation of caspase-3 expression.It provides a new method and choice for the diagnosis and treatment of lung cancer.