ABSTRACT
OBJECTIVE：To study the compositi on of the volatile oil from Compound chaihu guizhi decoction ，and to evaluate its in vitro anti-proliferative activity on human lung adenocarcinoma A 549 cells. METHODS ：The volatile oil from Chaihu guizhi decoction was extracted according to the steam distillation method of general rules 2004 in the 2015 edition of Chinese Pharmacopoeia（part Ⅳ）. The volatile oil components were analyzed by GC-MS combined with Kováts index ，and the relative content of each component was calculated by peak area normalization method. Using different concentrations of cisplatin （4，8， 16，32，64 mg/L）as positive control ，MTT assay was used to detect the inhibitory effects of different concentrations of volatile oil from Chaihu guizhi decoction （25，50，100，200，400 mg/L）on in vitro proliferation of A 549 cell after 48 h of treatment. Negative control group （with cells but without drugs ）was set up. RESULTS ：A total of 71 chemical components were isolated from the volatile oil ，among which there were 59 compounds identified ，sum of peak areas accounting for 84.99％ of the total peak area. The compounds with relatively high content included ar-curcumene （17.65％），β-bisabolene（9.57％），β-ocimene（7.05％）， α-curcumene（5.35％），2，5-dimethylbenzaldehyde（4.24％），linalyl isobutyrate （2.70％），α-cedrene（2.48％），δ-cadinene （2.07％）. Compared with negative control group ，the proliferation rate of cells were decreased significantly in 4-64 mg/L cisplatin groups and 25-400 mg/L volatile oil from Chaihu guizhi decoction groups （P＜0.05）. IC 50 of cisplatin and volatile oil from Chaihu guizhi decoction to in vitro proliferation of A 549 cells were 10.150 and 73.526 mg/L. CONCLUSIONS ：The volatile oil from Chaihu guizhi decoction mainly includes ar-curcumene ，β-bisabolene，β-ocimene，α-curcumene，which shows certain inhibitory effect on in vitro proliferation of A 549 cells.
ABSTRACT
Objective:To investigate the influence of cereblon(CRBN)on the inhibitory effect of thalidomide on the migration of human lung adenocarcinoma A549 cells and related mechanism. Methods: The effect of thalidomide on the A549 cell proliferation was assayed by the MTS method. CRBN in A549 cells was knocked down(A549CRBN cells) with RNA interference and lentivirus infection. Transwell assay was performed to detect the effect of thalidomide on the cell migration of A549 and A549CRBN cells. RT-qPCR assay was performed to detect the gene expression of N-cadherin and vimentin. Results: The thalidomide at the 1, 10, 50 and 100 μmol/L concentration did not affect the proliferation of A549 cells, while the 100 μmol/L thalidomide could significantly inhibit the migration of A549 cells. After the CRBN gene knockdown the migration ability of A549 cells was significantly decreased(P<0.01), and the gene expression of N-cadherin and vimentin was also significantly reduced in the A549CRBN cells. The inhibitory effect of thalidomide on the cell migration in the A549 cells disappeared after CRBN gene knockdown. Thalidomide could inhibit the gene expression of N-cadherin and vimentin in a CRBN-dependent manner. Conclusion: Thalidomide could inhibit the migration of A549 cells and the expression of the migration-related genes via CRBN.