| Human gastric cancer is one of the most common malignant tumor, the incidence of second place in the world. Over the past decadeWith the proto-oncogenes, tumor suppressor gene and the metastasis-associated genes of development in depth, it is recognized that many genetic changes involved in gastric cancer, including oncogene activation, inactivation of tumor suppressor genes and abnormal accumulation of growth factor and its receptor cell adhesion factor and other factors. The occurrence and development involved in a process and results of multi-factor, multi-step, multi-gene.At present, gastric cancer and precancerous lesions of different stages and different types of pathological changes in molecular biology and had undertaken extensive research. The topic for Apollon, Smac gene in gastric cancer specimens and cancer cell lines SGC-7901 cells were studied. Through the experimental study, we provide the new content of the occurrence of the tumor and the development of mechanisms. It is possible one kinds of new tumor markers and cancer treatment is expected to make it on a new target for the diagnosis and prognosis of tumor.Through the experimental sodium phenylbutyrate inhibited on human gastric cancer cell line SGC-7901. On the cellular level and sub-cellular level through the cell proliferation inhibition rate detection, flow cytometry, electron microscopy and RT-PCR detection, a preliminary conclusion that sodium phenylbutyrate inhibit on human gastric cancer SGC-7901 cell is confirmed. Apollon, Smac gene in gastric cancer cells expressed Separatly, suggesting that the Apollon, Smac gene may be related to the occurrence of malignant tumors.1.Experiment through the immunohistochemical method detect of the expression levels of Apollon.Results showed that: Apollon protein highly expressed in gastric cancer contrast to normal gastric tissue in the low-expression and the result exist of significant difference (p <0.05). Apollon protein expression unrelated to sex, age, incidence of the site. There is no significant difference between results (p> 0.05). Apollon protein expression relate to pathological staging and grading of gastric cancer .It unrelate to tumor size. In addition, Apollon protein expression in gastric cancer tissue relate to the degree of differentiation. Apollon protein expression higher in well-differentiated and undifferentiated relatively low and it has statistical significance (p <0.05). Apollon protein expression in the same relate to clinical stage.Lymph node metastasis may be nothing to do with and no significant difference exist between the two (p>0.05).2.Through the experimental method of RT-PCR we detect of Apollon, Smac gene in gastric cancer cases. Apollon mRNA in gastric cancer was highly expressed and it was significantly higher than adjacent tissues and normal gastric tissue (p <0.05). Apollon gene expression has nothing to do with the size of tumor (P> 0.05). With tumor differentiation, there is statistical significance in phases (P <0.05). It May be nothing to do with the lymph node metastasis. Smac gene in gastric cancer with low expression was significantly lower than adjacent tissues and normal gastric tissue (p <0.05), so we conclude the low expression of Smac gene relate to the occurrence of gastric cancer. Smac gene expression has nothing to do with the size of tumor (P> 0.05). With tumor type, differentiation degree, there is statistical significance (P <0.05). Tumor stage is worse when Smac is lower expression. However it is no statistical significance (p> 0.05). Lymph node metastasis relate to expression of Smac( P<0.05). The relation of expression of both Apollon and Smac gene are negatively correlated and Apollon high expression contrast to low expression of Smac in tumor.3. In this experiment through the MTT colorimetric assay ,we detect that sodium phenylbutyrate inhibit on human gastric cancer SGC-7901 cell proliferation. The results showed that with the dosage of sodium phenylbutyrate, gastric cancer cell inhibitory rate increased; With the use of extended time, the inhibitory rate of gastric cancer has also increased, showing time and dose-dependent manner. Through flow cytometry we found that sodium phenylbutyrate can make gastric cancer SGC-7901 cell block in the G0/G1 phase, so the proportion of G0/G1 phase cells increased. Through inverted microscope we observe cell morphology, negative control group of adherent growth SGC-7901 cells were polygonal. Uniform cell size, vigorous growth. After sodium phenylbutyrate role in cells, a relatively slow cell growth, cell expansion as well as the gap between the decrease in the number of cells showed significant cell growth inhibition. At the same time, cell morphology can be seen from the cobblestone into fibroblast-like cells similar to spindle shape.With increased drug concentration and the role of time, the change is obviousl. Under the electron microscope observation of SGC-7901 cells (control group, cells without drugs) were polymorphic, cell structure integrity, cell-developed, often very structured nuclear shape and nuclear-mass ratio increased significantly. SPB4.0mM apply for 72 hours after the SGC-7901 cells in the first days after 1,2,3, synaptic nuclei were round to reduce the nuclear and nuclear-mass ratio decreased and mitochondrial swell and cristae disappear .In cytoplasmic lipid droplets in the number of the increase in varying degrees, part of a large number of intracellular lipid droplets, lipid droplets of lipid for the synthesis of gastric epithelial cells to form lipid bodies in the electron microscope.The cytoplasm showed vacuolization body.This shows that benzene sodium butyrate inhibit the SGC-7901 cell.4. Colorimetric TUNEL System Dead EndTM detect different concentrations of SGC-7901 cell apoptosis in situ by sPB. Through optical microscope we observed cells stained and randomly selected 10 vision counting the number of positive cells. Observation results show that positive cells is not found in the control group in the field of vision, and in each group of drug positive cells can be seen, and with the increase in drug concentration, the number of positive cells increase. The results suggest that: sodium phenylbutyrate inhibit on growth of human gastric cancer cell.This experiment also confirmed that Apollon gene in gastric cancer cell lines express positively and its expression weaken when sodium phenylbutyrate role in human gastric cancer cell line.Smac gene exit in human gastric cancer cell line and express negatively, and With the sodium phenylbutyrate on human gastric cancer cell line Smac increased its expression and its expression changes relate to sodium phenylbutyrate on the end of the inhibition mechanism of gastric cancer.
|
PDF Full Text Request