Effect of different sequence administratation of elemene injection and gefitinib on human lung adenocacino?ma cell lines with different EGFR gene types

HUANG Wei; WANG Lin; YANG Aizhen; XU Haijun.

Abstract：analyze the antiproliferative and apoptotic effect of different sequences of combined elemene injec?tion and gefitinib on human lung adenocacinoma cell lines A549 and PC?9 with different epidermal growth factor receptor (EGFR) gene types in vitro, to further explore the optimal combination schedule of the two drugs. Methods Human lung adenocacinoma cell lines with wide?type EGFR gene(A549) and mutant?type EGFR gene(PC?9) were used in vitro in different experiment groups including con?trol group, elemene injection for 72 h group, gefitinib for 72 h group, elemene injection for 24 h sequence by gefitinib for 48 h group, gefitinib for 48 h sequence by elemene injection for 24 h group, gefitinib concomitant with elemene injection for 72 h group. MTT assay was used to measure cell proliferation. Cell apoptosis was analyzed by Annexin?V FITC／PI double marking staining assay and TUNEL assay. Results In A549 and PC?9 cells,the IC50 of elemene injection for 72 h were (27. 830±0. 614) μg／ml and (26. 461±2. 309) μg／ml respectively, the IC50 of gefitinib for 72 h were (8. 642±0. 261) μmol／L and (0. 044±0. 002) μmol／L respectively. In A549 cells, the proliferative inhibition rates of gefitinib (1,2,4,8,16 μmol／L) concomitant with elemene injection (5,10,20,40,80 μg／ml) were 32. 32%, 46. 47%, 56. 13%, 69. 13% and 81. 79%, which were higher than those in control group, elemene injection for 72 h group, gefitinib for 72 h group, elemene injection for 24 h sequence by gefitinib for 48 h group, and gefitinib for 48 h sequence by elemene injection for 24 h group with statistic significance ( P＜0. 01). In PC?9 cells, the proliferative inhibitory rates of gefitinib (0. 01,0. 02,0. 04,0. 08,0. 16 μmol／L) concomitant with elemene injection (5,10,20,40,80 μg／ml) were 36. 43%, 53. 24%, 61. 68%, 80. 74% and 86. 72%, which were also higher than those in the others( P＜0. 01). By Annexin?V FITC／PI double making staining assay, the apoptotic rates of gefitinib concomitant with elemene injection were (67. 4±4. 9)%,(67. 5±5. 6)% on A549 cells and PC?9 cells respectively, which were higher than those of the others( P＜0. 01). By TUNEL assay, gefitinib concomitant with el?emene injection had the highest apoptotic index on A549 cells and PC?9 cells, with (93. 48±8. 91)% and (85. 33±7. 62)% respec?tively. Conclusion The different sequences of combined elemene injection and gefitinib show the better antiproliferative and apoptotic effects on human lung adenocacinoma cell lines with different EGFR gene types in vitro. Gefitinib concomitant with elemene injection has the most significant effect.