Killing Effect Of B-Elemene With Cisplatin And Hyperthermia In Lung Cancer A549 Cells

BackGroud:Lung cancer is one of the malignant neoplasms, while none-small- cell lung cancer is one of the pernicious carcinoma which have bad prognosis. Chemotherapy shows unsatisfied effect for the multidrug-resistance. Hyperthermia is now applied in lung cancer therapy and used to treat carcinoma with chemotherapy. As a new antitumour drug,β-elemene shows better curative effect combined with chemotherapy in terminal lung cancer. But lack of adequate basis research limits the application ofβ-elemene to some extent.Objective:To observe the killing effect ofβ-elemene with cisplatin to A549 cell line at 37℃or 42℃in order to get a guiding concentration for drug application and attempt to investigate the possible mechanism.Methods:After treated with different concentration ofβ-elemene and cisplatin to A549 cell line at 37℃or 42℃, MTT assay was used to detected the cell inhibition rate. After treated with 15μg/ml or 60μg/mlβ-elemene and 4(ig/ml cisplatin at 37℃or 42℃, MTT assay was used to detected the cell inhibition rate. After treated with 4μg/ml cisplatin and 15μg/ml (or 60μg/ml) P-elemene at 37℃or 42℃, flowing cytometry was used to detect the cell cycles, apoptosis was observed by electronic microscope, RT-PCR and Western Blots were applied to detect mRNA transcription and protein expression of HSP70 and MDR1, while Bax/Bcl-2 mRNA detected by RT-PCR. Results:MTT assays indicated no significant difference between the cell inhibition rate ofβ-elemene and cisplatin at 37℃or 42℃. Inhibition rate at 42℃of 4μg/ml cisplatin with 15μg/ml P-elemene was higher than that at 37℃and showed significant difference, but inhibition rate at 42℃of 4μg/ml cisplatin with 60μg/mlβ-elemene was lower than that at 37℃. No significant difference was observed among HSP70 mRNA transcription. HSP70 protein expressions were mostly up-regulated and there were significant difference. MDR1 mRNA transcriptions and protein expressions were mostly down-regulated. After treatment with cisplatin 4μg/ml and 15μg/mlβ-elemene in 42℃it was up-regulated(compared with 37℃control, or 4μg/ml cispaltin and 15μg/ml P-elemene in 42℃, P<0.05) but there were no significant difference among the MDR1 protein expressions. Bcl-2 mRNA transcriptions were mostly down-regulated and was observed significant difference, while Bax mRNA were observed no significant difference.Conclusion:At 37℃or 42℃,β-elemene with cisplatin may result in diverse outcomes. The increased inhibition of 15μg/mlβ-elemene with 4ug/ml cisplatin at 42℃may mainly owe to the lower expression of Bcl-2 gene and the reduce of drugresistance.