| Background and ObjectiveGastric carcinoma is one of the commonest carcinoma in our province and country. It was a complicated course that multiple factors, many steps and genes participated in generation and development. Many researches expressed that generation and development of gastric carcinoma correlated not only with oncogenes and anti-oncogenes, but also with related genes of apoptosis.The human programmed cell death 5 (PDCD5) was a new human genes named TFAR19(TF-1 cell apoptosis related gene 19) which was first reported in the international by center for human disease genimics of Peking University in 1999. It was a new apoptosis-related gene cloned form human leukemia cell line TF-1 PDCD5 expressed in more than fifty human tissues, especially in the heart, testis, kidney, adrenal gland and placenta. Remarkably, the expression of PDCD5 mRNA in fetal tissues is significantly lower than that in adult tissues. PDCD5 had no expression or low expression in many diseases, particularly in cancer.Many researches showed that PDCD5 promoted a variety of apoptosis, in the increased expression of apoptosis. PDCD5 exogenous certain way by importing a variety of cell lines, their biological characteristics of the cells did not have a tangible impact on its performance alone low. However, if the use of chemotherapy drugs to deal with, TRAL treatment, the removal of cytokines, the removal of serum, irradiation, anti-FAS antibody treatment caught significantly promoting apoptosis. Results showed that the PDCD5 was an effective promoter of apoptosis, rather than apoptosis-inducing agent.Resveratrol (Res) was a naturally accruing phytoalexin polyphenolic compound found in various plants, including grapes, polygonum, and peanuts, and had been shown to exhibit a wide range of pharmacological properties and was believed to play a role in the chemoprevention and treatment of human cancer. It caught significantly inhibiting a variety of human tumor cell growth and induced apoptosis.Effects of resveratrol on expressions of PDCD5 protein and mRNA in human gastric adenocarcinoma BGC-823 cell and the role of growth inhibition on BGC-823 cell had been studied by our research group. In order to research the effect of resveratrol on the expression of PDCD5 protein and mRNA in human gastric adenocarcinoma cell, nude mice model with transplanted human gastric adenocarcinoma was built.Immunohisr of human gastric adenocarcinoma; and in situ hybridzation was used to examine PDCD5 mRNA expression in transplanted tumor of human gastric adenocarcinoma; and TUNEL method to examine apoptosis of transplanted tumor of human gastric adenocarcinomtochemistry SP was used to examine PDCD5 protein expression in transplanted tumoa.Materials and Mmethods1. Samples and building nude mice model with transplanted human gastric adenocarcinoma cells: Human gastric poor differentiated adenocarcinoma cell BGC-823 was purchased from Chinese Academy Science Shanghai cell biology graduate school. Through recoverying incubation and culturing, the BGC-823 cells were handled by 200μmol/L resveratrol for 72h, and set up blank control groups (?)medium control group), cisplatin control groups (3μg/ml), combination groups (?)sveratrol 200μmol/L + cisplatin 3μg/ml), and the human gastric adenocarcinoma BGC-823 cells were injected into the skin of nude mice model.2. Immunohistochemistry SP was used to examined PDCD5 protein expression in transplanted tumor tissue of human gastric adenocarcinoma3. The mRNA expression of PDCD5 in transplanted tumor tissue of human gastric adenocarcinoma was examined by in situ hybridzation.4. TUNEL method was used to detect the cell apoptosis of transplanted tumor of gastric adenocarcinoma.5. Statistical analysis: All the data were analyzed by SPSS 11.0 statistical softwarepackage, the data were expressed by mean±standard deviation ((X|-)±S), and analyzed using the t-test for two groups and ANVOA for more than two groups. The level of significance was a=0.05.Results1. The positive expressions of PDCD5 protein and mRNA in transplanted tumor of gastric adenocarcinoma: The positive expression of PDCD5 protein was located in cytoplasm, showed brown color. The positive expression of PDCD5 mRNA in transplanted tumor of gastric adenocarcinoma was also located in cytoplasm, but showed amethyst color.2. Immunohistochemistry results: The expression of PDCD5 protein was significant higher in group by resveratrol with 200μmol/L than that in blank control groups, there was significant difference between them (P<0.05), but lower than in cisplatin control groups, and combination groups. There was a significant difference between them (P<0.05).3. In situ hybridization results: PDCD5 mRNA expressed significant higher in group by resveratrol with 200μmol/L than that in blank control groups, there was significant difference between them (P<0.05), but lower than in cisplatin control groups, and combination groups. There was a significant difference between them (P<0.05).4. TUNEL results: The apoptotic cells showed nuclei brown, chromatin distribution unevenly and dyeing depth varies detected by TUNEL method. Blank control groups almost did not show apoptotic cells and in experiment groups the number of apoptotic cells increased with the dosage of resveratrol increased, compared between the experiment groups, there were significant differences (P<0.05).Conclusions1. Resveratrol can up-regulate the expression of PDCD5 protein and mRNA in transplanted tumor of gastric adenocarcinoma, it indicates that resveratrol induce apoptosis in nude mice model with transplanted human gastric adenocarcinoma cells through up-regulating the expression of PDCD5 protein and mRNA.2. Effects that up-regulating PDCD5 protein, mRNA and inducing apoptosis are the strongest in combination groups, it indicates that resveratrol can enhance the effect of cisplatin induce apoptosis. |
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