Effects Of Lycorine And Lycorine Hydrochloride On Cell Proliferaion And Apoptosis In A549 And Eca-109 Cells

Malignant tumor is a serious disease harm to human health currently. Anti-tumor chemicals in oncotherapy also produce toxic and side effect to normal cells of human body. Therefore, from the natural plant to find efficient low toxic anti-tumor active ingredient is research highlight in recent years at home and aboard. Lycorine(LH) and lycorine hydrochloride(LH) extracted from Amaryllidaceae, have been reported with good anti-tumor efficacy in the leukemia cells(HL 60 and K562), multiple myeloma cells(KM3), ovarian cancer cells(Hey1B), non hormone-dependent prostate cancer cells(PC3 M, DU145, LNCa P, 22 RV1). But their anti-tumor mechanism is not so identical.In this report we select lycorine(LH) and lycorine hydrochloride(LH) to investigate the molecular mechanism of cell proliferation inhibtion and cell apoptosis induction in lung cancer A549 and esophageal Eca-109 cells.The main researches are as follows:(1) Cell viability of A549/Eca-109 cells was detected by MTT method after induction of0,2.5,5.0,10.0,20.0μM LY/LH for 24 h and 48 h.(2) Cell morphology of A549/Eca-109 cells was observed by fluorescent microscope after induction of 0,2.5,5.0,10.0,20.0μM LY/LH for 24 h.(3) Cell apoptosis rate of Eca-109 cells was detected by flow cytometer after induction of0,1.2, 2.4,4.8μM LH for 24 h.(4) Cell cycle changes were analysed by cytometer after induction of 0,2.4,4.8μM LH for 24 h.(5) p53, Bcl-2, Bax, Caspase-3, Caspase-9 mRNA transcription level were analysed by qRT-PCR method after induction of 10μM LH for 24 h.(6) p53,Bcl-2,Caspase-9,Cyclin D1 gene expression and phosphorylation level of NF-κB and GSK-3β were analysed by western blot after induction of 0,0.5,1.0μM LH for6 h.The results are as follows:(1) LY and LH inhibited cell viability of A549/Eca-109 cells in a time and dosedependent manner.(2) After being treated with a certain concentration of LY/LH,cells were stained with AO/PI.Cell morphology was observed with red fluorescence and increased apoptotis bodies under fluorescence microscopy.(3) After being treated with 0,1.2,2.4,4.8μM LH for 24 h,cell apoptosis rate of Eca-109 cells increased in a dose-dependent manner.(4) After being treated with 0,2.4,4.8μM LH for 24 h,the number of Eca-109 cells in the G1 phase increased in a dose-dependent manner.(5) After being treated A549/Eca-109 cells with 10μM LY/LH for 24 h, qRT-PCR results showed mRNA transcription level of p53,Bax,Caspase-3,Caspase-9 up-regulated obviously while Bcl-2 down-regulated.(6) After being treated Eca-109 cells with 0,0.5,1.0 μM LH for 6h,western blot results showed expression of p53 and Caspase-9 increased while Bcl-2 and Cyclin D1 decreased in a dose-dependent manner.Phosphorylation level of SAPK/JNK and GSK-3βincreased in a dose-dependent manner.In inclusion, the results demonstrated that Lycorine(LH) and lycorine hydrochloride(LH)could inhibit cell viability of lung cancer A549 cell and esophageal cancer Eca-109 cells,and induce apoptosis.Our results will provide theoretic foundation for LY and LH as new natural antineoplastic drugs.