Effects Of Curcumin Combined Paclitaxel On Biological Effect Of Human Lung Adenocarcinoma Cell Line A549

Objective:The aim of this study was to invesigate the effects of curcumin combined paclitaxel on biological effect of human lung adenocarcinoma cell line A549 in vitro and explored possible mechanism of this effection.Methods:(1)The culture of A549 human lung cancer cells:it was cultured in the 5%C02,37? and saturated humidity constant temperature incubator with RPMI1640 medium containing 10%fetal bovine serum.(2)Experimental group and treatment:this study setted the control group,DMSO group,curcumin group,paclitaxel group and the curcumin combine paclitaxel group.When the cells grow to logarithmic period,the experimental groups were treated with no drug,DMSO that the concentration same as curcumin group,20?mol/L curcumin,4nmol/L paclitaxel,20?mol/L curcumin and 4nmol/L paclitaxel respectively.(3)Detected the paclitaxel and curcumin IC50 of A549 human lung cancer cells for 48h:when the A549 human lung cancer cells grew with adherence for 24 hours after inoculated in 96-well plate,respectively add different concentration of paclitaxel for Onmol/L,0.5nmol/L,lnmol/L,2nmol/L,4nmol/L,8nmol/L;and add curcumin for Oumol/L,5umol/L,10umol/L,20 umol/L,40 umol/L,80 umol/L.Then the IOD value was detected and IC50 was calculated after treating for 48h.(4)The MTT test:then the IOD value was detected and inhibition ratio for each experimental group was calculated after treating for 48h.(5)Detection of cell cycle distributions and apoptosis:The cell cycle distributions and apoptosis was detected by flow cytometry after treating for 48h.(6)The migrative distance was measured when each group was treated for 48h after the cells proliferated to form a full layer.(7)Each group was treated for 48h after Serum-free cell suspension was injected in upper compartment of transwell,then counted the number cells invading through the Matrigel of each group after dye.(7)The expression of HIF-1? and VEGF were detected by ELISA method for each group after treating for 48h.Results:(1)the IC50 of paclitaxel was 4 nmol/L for A549 human lung cancer cells after treating for 48h,the IC50 of curcumin was 20 umol/L.(2)The proliferation of A549 human lung cancer cells for combined group was significantly inhibited and the inhibition rate is significantly higher than control group,paclitaxel group and curcumin group.Curcumin and paclitaxel has a synergistic effect for growth inhibition of A549 lung cancer cells,the value of Q was 1.072338.(3)There was no significant difference between the DMSO group and the control group for cell cycle distribution(P>0.05).The cell cycle was blocked in S phase and G2/M phase for the curcumin group and in G2/M phase for the paclitaxel group and in G0/G1 phase and S phase for the combined group.(4)The apoptosis rate of the DMSO group has no significant difference with the control group(P>0.05).The apoptosis rate of experimental groups were higher than the control group,which of the combined group is higher than other groups,the difference was statistically significant(P<0.05).(5)Compare with the control group,the migrative distance and the number of cells invading through the Matrigel of the curcumin group,the paclitaxel group and the combined group were significantly shorter and less,the difference was statistically significant(P<0.05).The difference of that between the curcumin group and the paclitaxel group was no statistical significance(P>0.O5).Which of the combined group was significantly shorter and less than the curcumin group and the paclitaxel group,the difference was statistically significant(P<0.05).(6)The expression of HIF-1? and VEGF for the DMSO group has no significant difference with the control group(P>0.05).Compare with the control group,which of the curcumin group,the paclitaxel group and the combined group were significantly decreased,the difference was statistically significant(P<0.05).The difference of the expression level of HIF-1? and VEGF between curcumin group and paclitaxel group was no statistical significance(P>0.05).Compare with curcumin group and paclitaxel group,the HIF-la and VEGF expression level of the combined group was significantly down-regulation,the difference was statistically significant(P<0.05).The expression levels of VEGF were positively correlated with the HIF-1?expression level(R = 0.976,P<0.05).Conclusion:1)Curcumin combined paclitaxel can significantly promote apoptosis and inhibit the proliferation,invasion and metastasis of A549 human lung cancer cells,there is a synergistic effect for A549 human lung cancer cells treated with this two dugs.(2)Curcumin combined paclitaxel can significantly decreased the expression of HIF-la and VEGF and the effects of curcumin combined paclitaxel on biological effect of A549 human lung cancer cells in vitro may be connected with down-regulation of HIF-1? and VEGF expression.