| Cancer is a somatocyte disease that is characterized by out of control and cells unrestrained proliferation, and it heavely harm to people's health. The methods of cancer therapy include surgical operation, pharmacotherapy and radiation, etc. Currently, the new researching of anti-tumol/Lor drugs has become a hotpot in this field. Many natural and synthetic medicines play important roles in prevention, treatment and rehabilitation of the cancer. Hebei province is a high incidence area of esophageal cancer. Our laboratory has been going in for developing new anti-tumol/Lor drugs for some years. Lupeol, a kind of Triterpenes compound, was abstracted from some food and anti-tumol/Lor herbs. It was confirmed that Lupeol can inhibit many cancer cells proliferation, including lung cancer, prostate cancer, melanoma and leukemic cells in vitro. In the process of studying anti-tumol/Lor effect of Lupeol for humol/Lan esophageal carcinoma cells, we found that Lupeol can apparentely inhibit TE-1 cell proliferation in vitro. In the further study, we explored the mechanism of Lupeol inhibition TE-1 cell proliferation, providing the theory base of Lupeol for clinical development.Partâ… The study for role and mechanism of Lupeol induced TE-1 cells apoptosis in vitroObjective: Detecting the effect of Lupeol inhibition TE-1 cells proliferation, to observe the morphologic and ultrastructural changes of TE-1 cells in vitro and to measure the expression level of genes and proteins in TE-1 cells treated by Lupeol.Methods:1 MTT assay was used to detect the inhibitory effect of Lupeol(25, 50, 100, 150 and 200μM)on humol/Lan esophageal carcinoma cells TE-1 for 24, 48 and 72h.2 Observe the morphologic changes of TE-1 cells (50, 100 and 150μM Lupeol for 24h) using inverted phase contrast microscope after Giemsa's staining.3 Observe the ultrastructural changes of TE-1 cells after treatment with 150μM Lupeol for 48h by transmission electron microscope (TEM).4 The expression levels of Caspase-3, -8, -9, Fas, Bax and Bcl-2 mRNA were detected by RT-PCR after TE-1 cells were treated by 50, 100 and 150μM Lupeol for 48h.5 The proteins expression of Fas, Cyt-c, Bax and PARP in TE-1 cells after treatment by 50, 100 and 150μM Lupeol for 48h were detected by western blotting.Result:1 Lupeol inhibited the proliferation of humol/Lan esophageal carcinoma cells TE-1 in a dose dependent manner after TE-1cells were treated by 25, 50, 100 and 150μM Lupeol for 24 and 48h (P<0.01).2 It was observed cells growth sparseness, loss of volumol/Le, cytoplasm concentration, karyokinesis and deformation into round in the experiment groups. By the contrast, the cells in control group were observed appearance regularity, growth intensively. They maintain polygon shapes.3 It was observed the ultrastructural organization changes of TE-1 cells using transmission electron microscope(TEM). Cell membrane increased thickness, chromatin highly concentrated, collected under karyolemma and thickness unevenness. It formed crescent, part of karyolemma formed acute angle, tuber to cytoplasm, some vacuole was in intracytoplasm. The cells in control group were observed a lot of microvillus in cells surface and some mitochondria, rough endoplasmic and free ribosomes intracytoplasm.4 RT-PCR result demonstrated that expression of Caspase-3, -8, -9, Fas, Bax mRNA in experimental group cells were increased compared with that in control group, but Bcl-2 mRNA decreased (P<0.05).5 western blotting result indicated that the expression of PARP, Fas, Bax and Cyt-c proteins in experimental groups were increased compared with that in control group.Partâ…¡The study for mechanism of Lupeol on TE-1 cells cycle arrestObjective: To detect effect of different concentration of Lupeol on TE-1 cell cycle and its mechanism from the level of gene and protein of TE-1 cells.Method:1 After TE-1 cells were treated by 50, 100 and 150μM Lupeol for 48h, the cycle distribution and apoptotic rate were detected by flowcytometry (FCM).2 western blotting method was used to analyse the expression change of CyclinD1 and CDK4 proteins in TE-1 cells treated by 50, 100 and 150μM Lupeol for 48h.3 Expression change of p53 mRNA was detected by RT-PCR method after TE-1 cells were treated by 50, 100 and 150μM Lupeol for 48h.Result:1 It was demonstrated that the cells of G1 phase in experiment group (50, 100 and 150μM)were increased, as compared with that in control group, apoptosis rate in experiment group cells was increased compared with that in control group (P<0.05).2 western blotting result displayed that expression of CyclinD1 and CDK4 proteins were decreased in experimental groups (P<0.05).3 RT-PCR result was showed that expression of p53 mRNA was increased in experimental groups than control group (P<0.05).Conclusion:Lupeol can induce TE-1 cell apoptosis and cell cycle arrest, the mechinasm of which were related with down-regulation of CyclinD1 and CDK4 proteins and up- regulation of p53 mRNA.
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