Studies On The Inhibitory Effect Of Lycorine On Proliferation Of Lung Cancer NCI-H460 Cells And Growth Of Lewis Lung Cancer Bearing Mice

Objective: The author studied the inhibitory effect of lycorine on proliferation of human lung cancer NCI-H460 cells and its effect on NCI-H460 cell apoptosis,and in vivo anti-tumor effect on Lewis lung cancer bearing mice,and investigate the anti-tumor mechanism of lycorine,and find a new efficient and low toxicity chemotherapy drugs in the clinical treatment of non-small cell lung cancer.Methods: 1.The cytotoxic effect of lycorine on NCI-H460 cells:(1)The inhibitory effect of lycorine on NCI-H460 cells was detected by MTT assay.(2)The apoptosis rate of NCI-H460 cells was detected by flow cytometry.(3)The activity of a apoptosis-related factor Caspase 3 was tested by colorimetric assay.2.Antitumor effect of lycorine on Lewis lung cancer mice:(1)to cultivate of Lewis lung cancer cell lines(2)Establishment of mouse Lewis lung cancer xenograft model(3)The mice were sacrificed and the tumor inhibition rate,spleen index and thymus index were calculated(4)HE staining to observe the cell morphology.Results: 1.NCI-H460 cells were treated with lycorine for 24 h,and the morphology of NCI-H460 cells was irregular,the cells were shrunk,and the size of cells was different.The degree of cell fragmentation increased with the dose increased,the number of cells gradually decreased.2.The proliferation of NCI-H460 cells was significantly inhibited by NCI-H460 cells treated with lycorine for 24 h,which was significantly different from that of the control group(p<0.05,p<0.01),and showed a certain concentration correlation.IC50 was 5.79±0.11 μmol/L,indicating that lycorine could inhibit the proliferation of lung cancer NCI-H460 cells.3.With the increase in the concentration of lycorine solution,The apoptotic rate of the treated group was significantly higher than that of the saline group(p<0.01),The highest concentration of 16 μmol/L,the apoptotic rate reached 46.2 ± 3.46%.The results indicated that lycorine had an induction effect on the apoptosis of NCI-H460 cells and showed a dose-dependent manner.4.With the increase of lycorine concentration,the activity of Caspase 3 increased gradually.Compared with the saline control group,the activity of Caspase 3 was significantly increased(p<0.01).The results showed that lycorine could enhance the activity of Caspase 3 in NCI-H460 cells,and induce NCI-H460 cell apoptosis.5.The results showed that compared with the saline group,the low,medium low,medium,medium high and high dose groups of lycorine inhibited on the tumor growth of Lewis lung cancer bearing mice.The effect of high dose group(40mg/kg)was the most significantly,the tumor inhibition rate was 56.8%(p<0.01).6.The results of HE staining of solid tumor tissue in tumor-bearing mice were as follows: the tumor cells were grown in the normal saline group,and the nucleus was less compacted and had no necrosis or punctate necrosis.The dosage group of lycorine and cyclophosphamide group tumor cells showed a large number of nuclear pyknosis,nuclear fragmentation,the number of tumor cells decreased,arranged in loose.It showed the most significant changes especially in high dose group(40 mg/kg),the tumor cells were scattered in the distribution,the number decreased significantly.Conclusions: Lycorine in vitro can effectively inhibit the proliferation of NCI-H460 cells and promote its apoptosis,the mechanism may be related to activation of the expression of Caspase 3 to induce apoptosis of tumor cells;Lycorine in vivo can also effectively inhibit the growth of tumor on Lewis Lung cancer cell bearing mice.