| Objective: Observation the Euphorbia lunulata Bge extract in vitro proliferation inhibition of human gastric cancer cell line SGC-7901; Discussion Euphorbia lunulata Bge ability to induce human gastric cancer SGC-7901 cell apoptosis and analyze the mechanisms of its possibleMethods :Using in vitro culture techniques, Euphorbia lunulata Bge in different concentration(0.2 g/L, 0.4 g/L, 0.8 g/L, 1.6 g/L, 3.2 g/L) to act on human gastric cancer SGC-7901 cells, the effects of MTT with different concentration and Euphorbia lunulata Bge assay on the proliferation of human gastric cancer SGC-7901 cells; apoptosis changes of SGC-7901 cells were observed by fluorescence microscope; the apoptosis and apoptosis of SGC-7901 cells were detected by flow cytometry; the expression of Bcl-2 protein in gastric cancer SGC-7901 cells was detected by immunohistochemistry.Results :1.Determination of different concentrations of Euphorbia lunulata Bge to SGC-7901 gastric cancer cells 24 h, 48 h, 72 h was significantly inhibited by MTT, compared with the control group, there were significant differences(P<0.05), and with the prolongation of time, the inhibiting effect produced more obvious, showing time and dose dependent.2.A phase contrast microscope at different concentrations of Euphorbia lunulata Bge treated SGC-7901 cells in experimental group compared to the control group, cells changed obviously, morphological growth slow, the adherent state difference, cell edge is not the whole, shrinkage, and turned round, poor shape, loss of normal cell morphology.3.To observe the effect of different concentrations of Euphorbia lunulata Bge treated SGC-7901 cells under the fluorescence microscope, in contrast to the control group compared with the control group, the experimental group cells cell growth is slow andsparse, cellvolume becamesmall, round, nuclear pyknosis, cytoplasm concentrati on; refractive becomes strong, showing an irregular shape, a strong compact fluorescent, DNA close to the nuclear membrane and concentrated, with a small amount of apoptotic body formation.4.Flow cytometry cell cycle analysis showed, cells through after drug treatment,cells decreased cell number in the G1 and S phase, and increase in cell number of G2/M phase, speculated that Euphorbia lunulata Bge may inhibit the gastric cancer SGC-7901 cells in G2/M, induced SGC-7901 cells apoptosis, and cell cycle arrest effect andapoptosis rate were dose dependent.5.Immunohistochemical results showed that, the expression of Bcl-2 was SGC-7901 in cytoplasm, many brown granules, more strong positive cells, brown yellow granules increased, most strong expression, with the increase of drug concentration, the expression of Bcl-2 protein in SGC-7901 cellsincreased with time, dose and positive expression rate significantly reduced, colorationfades, strong positive staining to reduce the proportion. Tan no expression or weak expression.Conclusion : 1.MTT results showed that in a certain range, the proliferation of Euphorbia lunulata Bge can inhibitgastric cancer SGC-7901 cells, was significantly difference compared with control group(P<0.05), and with the dose increased and time prolonged, the rate of apoptosis is more obvious2.Observed by inverted phase contrast microscope and fluorescence microscope, cell morphologychanged obviously. Under the microscope, can clearly see the apoptotic bodies, confirmed the Euphorbia lunulata Bge extract can induce apoptosis of SGC-7901 gastric carcinoma cells3.FCM showed that different concentrations of Euphorbia lunulata Bge to the changes of cell cycle, experiments show that the G2/m phase of the cell cycle inhibition ratio increased significantly, that Euphorbia lunulata Bge can block the SGC-7901 cell cycle arrest in G2/M phase and induce its apoptosis, the rate of apoptosis in a dose and time dependent manner.4.Immunohistochemistry showed that different concentration of Euphorbia lunulata Bge effect on human gastric cancer SGC-7901 cells, results showed that the expression of Bcl-2 protein in SGC-7901 cells, the expression of apoptosis may be related to the down-regulation of Bcl-2 Euphorbia lunulata Bge, as one of the inhibition apoptosis gene and down-regulation of Bcl-2 gene may be an important mechanism of inducing apoptosis... |
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