| Objective: Lung cancer is one of the highest mortality of malignant tumor in the world. The rate of non-small cell lung cancer incidence in lung cancer incidence is more than 80%. And metastasis is the leading cause of death from lung cancer. Therefore, preventing the transfer of lung cancer cell is effective measures to control the progression of tumors, improve the quality of patient life and prolong patient life. In recent years,cryptotanshinone as a major ingredient of Salvia,its anti-tumor activity was highly valued. This subject chose two kinds of human non-small cell lung cancer lines A549 and H460, which first detected the expression of integrins and E-cadherin, and then assess the effect of cryptotanshinone on cell growth, migration, invasion, and the cell cycle of non-small cell lung cancer cells. This subject explore the role of integrin and E-cadherin in non-small cell lung cancer metastasis process, which provides new targets for the treatment of non-small cell lung cancer, and lay the theoretical foundation for Cryptotanshinone in treatment of lung cancer. Methods:(1) Flow cytometry was used to detect the expression of E-cadherin and β integrin in A549 cells and H460 cells.(2) A549 cells and H460 cells were treated with different concentrations(5, 10, 15, 20, 30, 40, 60 μmol/L) of cryptotanshinone in vitro for 24 h. Cell proliferation was analyzed by MTT assay.(3) After treated with different concentrations(10, 30, 50 μmol/L) of cryptotanshinone for 24 h of A549 cells and H460 cells, cellular migration was evaluated by wound scratch assay and transwell experiments.(4) A549 cells and H460 cells were treated with cryptotanshinone(40μmol/L) for 48 h. Flow cytometry was used to detect the change of cell cycle and the expression of E-cadherin and β-integrin. Results:(1) MFI of β-integrin in A549 cells is significantly higher than H460 cells, but a few differences in the expression of E-cadherin. Then we obtained higher-expression tumor lines A549 and lower-expression of tumor lines H460.(2) MTT assay showed that cryptotanshinone had effective inhibition of proliferation in concentration of 5μmol/L to the two cells, and with the increase of drug concentration, cell viability decreased significantly. When the doses were higher than 20 μmol / L, no significant effect appears. IC50 was(31.59 ± 1.25) μmol/L for A549 cells and(42.45 ± 0.75) μmol/L for H460 cells. Cryptotanshinone showed significant inhibition in A549 cells.(3) Wound scratch assay showed that cell scratch healing slowly when increased with the concentration cryptotanshinone. Cell migration was significantly inhibited. Compared with the control group, scratches width was significant difference.(4) Transwell experiments showed that after treated with cryptotanshinone, the migration of A549 and H460 cells was decreased significantly. Results of A549 cells: the number of cells in the control group was 129.45 ± 14.26, experimental group cells were 85.36 ± 11.22(10μmol/L), 45.82 ± 7.32(30 μmol/L), 36.21 ± 3.12(50 μmol/L). Results of H460 cells results: the number of cells in the control group was 145.35 ± 13.48, experimental group cells were 96.25 ± 13.46(10 μmol/L), 55.35 ± 8.42(30 μmol/L), 45.21 ± 5.32(50 μmol/L). With dose of cryptotanshinone increases, the number of permeable membrane cells reduced. Cryptotanshinone showed higher inhibition rate of migration to A549 cells(5) Data from flow cytometry assay indicated that cryptotanshinone arrested cell cycle in G1 phase, rised proportion of cells in G1 phase and decreased proportion of cells in S phase. Furthermore, cryptotanshinone increased expression of E-cadherin on the surface of cells, while decreased expression of β-integrin. Conclusion: This study first selected high and low expression cell of integrin,found the ability of that cryptotanshinone inhibited cell proliferation, migration and invasion in high integrin-expression cell is stronger. The mechanism may be related to cryptotanshinone effect on upregulation of E-cadherin, downregulation of β-integrin and arrest of cell cycle. Key words: cryptotanshinone;non-small cell lung cancer;flow cytometry assay;... |
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