Study On Curcumin Increasing The Sensitivity Of A549 Cells To Paclitaxel And Reducing Paclitaxel-induced Hepatotoxicity

AIM: Because of the drug resistance and adverse reactions,paclitaxel has a poor therapeutic effect on non-small cell lung cancer(NSCLC).It has been reported that curcumin is one of the effective drugs to enhance the chemotherapy effect of various tumor chemotherapy drugs.So this experiment studied on curcumin increasing the sensitivity of A549 cells to paclitaxel and reducing paclitaxel-induced hepatotoxicity.METHODS:(1)A549 cells(Non-small cell lung cancer cell)and L-02 cells(human normal liver cell)were cultured in vitro,and the cells were sub-cultured in the logarithmic growth phase.Each cell was intervened for 48 hours according to the following grouping: control group(without any drug,only cell culture solution),curcumin group(2.5,5,10,20,40 ?mol/L curcumin solution was added in turn,set this group only for A549 cells),paclitaxel group(0.06,0.125,0.25 ?mol/L paclitaxel solution was added in turn,combination group(5 ?mol/L curcumin + 0.06 ?mol/L paclitaxel;5 ?mol/L curcumin + 0.125 ?mol/L paclitaxel;5 ?mol/ L curcumin + 0.25?mol /L paclitaxel).CCK-8 method was used to detect the cell proliferation of each group,and screening the appropriate concentration of the two drugs in combination for subsequent experiments(5?mol/L curcumin + 0.125 ?mol/L paclitaxel).(2)According to the concentration of curcumin and paclitaxel selected above,the experiment was divided into four groups: control group(without any drug,only cell culture solution),curcumin group(5 ?mol/L curcumin solution),paclitaxel group(0.125 ?mol/L paclitaxel solution),combined group(solution consisting of 5?mol/L curcumin + 0.125 ?mol/L paclitaxel).A549 cells and L-02 cells were intervened for 48 h,respectively.Flow cytometry was used to detect the apoptosis of each cell.(3)Western blotting was used to detect the expression levels of STAT3,Bcl-2 and Bax in A549 cells treated according to the above-mentioned groups.(4)For the L-02 cells that were intervened according to the above group,the concentrations of ALT and AST in the supernatant of each group of L-02 cells were detected by ELASE method.RESULTS: 1.The results of CCK-8 showed that:(1)Curcumin and paclitaxel could inhibit the proliferation of A549 cells in a concentration-dependent manner.In another,compared with the single-drug group,the combination group was more effective in the inhibition of the proliferation of A549 cells(p < 0.05).(2)Paclitaxel also inhibited the proliferation of L-02 cells,and as the drug concentration increased,the rate of inhibition of L-02 cells increased.However,compared with the equivalent dose of paclitaxel group,the inhibition rate of L-02 cell proliferation was significantly decreased by the comparison group(p < 0.05).2.The results of flow cytometry showed that:(1)Compared with the the curcumin group and the paclitaxel group,the apoptosis rate of A549 cells was significantly increased in the combination group(p < 0.05).(2)The apoptotic rate of L-02 cells in paclitaxel group was significantly higher than that in the control group(p < 0.05).But the apoptosis rate of the combination group was obviously lower than the equivalent dose of paclitaxel group(p < 0.05).3.Western blotting results showed that compared with the control group,the expression of STAT3 and Bcl-2 in A549 cells of curcumin group and paclitaxel group was decreased,and the expression of Bax was significantly increased(p < 0.05).Compared with the single-agent group,the expression of STAT3 and Bcl-2 was decreased and the expression of Bax was significantly increased in A549 cells of combination group(p < 0.05).4.The results of ALT and AST detected by ELASE showed that the concentrations of ALT and AST in the supernatant of L-02 cells in the paclitaxel group were significantly higher than those in the control group,and the results were significantly different(p < 0.05).The levels of ALT and AST in the supernatant of the combination group were significantly lower than the same dose of paclitaxel group(p < 0.05).CONCLUSION:(1)Both curcumin and paclitaxel can inhibit proliferation and promote apoptosis of A549 cells,and they all shown a trend of concentration-dependent.The combination of low-dose curcumin and paclitaxel was more effective than single drug,which suggested that there existed a synergistic effect between curcumin and paclitaxel on proliferation and apoptosis of A549 cells.And the mechanism of apoptosis of A549 cells may be related to down-regulation of STAT3,Bcl-2 and up-regulation of Bax.(2)Paclitaxel had the effects of proliferation inhibition and apoptosis promotion on L-02 cells,and the effect was more obvious with the increase of drug concentration.Compared with the combination group of low-dose curcumin and paclitaxel,the same dose paclitaxel group can inhibite cell proliferation and promote apoptosis more effectively,and the levels of ALT and AST in the supernatant were significantly higher than those in the combination group(p < 0.05),suggesting that low dose of curcumin can alleviate liver damage caused by paclitaxel.