| Objective:Observe proliferation and apoptosis effects of different radiation activity125I seeds on nude mice subcutaneous human poorly differentiated gastric carcinoma (BGC823) transplantation tumor cell, Explore on nude mice subcutaneous human poorly differentiated gastric cancer cells proliferation inhibition of the biological mechanism, Compare with inhibition effect of different radiation dose of125I seeds organizational implantation on human poorly differentiated gastric cancer.Method:Human poorly differentiated gastric cancer cell line (BGC823) were cultured, and when cells are in a proliferative phase it was grown in BLAB nu/nu nude mice(70) subcutaneous part. Raised3weeks,64nude mice that their transplantation tumor diameter was between0.6cm with1.0cm were selected as an animal model, and they were randomly divided into4groups, A/B/C/D (n=16), group A as control group (0mCi)(1mCi=37Mbq), was implanted in blank seed, and B/C/D group as the experimental groups were implanted in the radiation activity1251seeds for0.4mCi/0.6mCi/0.8mCi. After seeds implantation, Measured the length of the tumor size, every3days, calculated tumor volume and rendering tumor curve of growth. After seeds implanted, in14days/28days, according to the order of execution of the control group and the experimental group in tumor-bearing nude mice,8mice per group were killed, stripping the tumor body. Light high power microscopy randomly select3visual field count tumor cells and apoptotic cell number, counting the apoptosis index (Apoptosis Index, AI), AI=the number of apoptotic cells divided by counting the number of tumor cells x100%. By TdT-mediated dUTP-biotin nick end labeling (TUNEL) method detected of tumor cell apoptosis level; and by immunohistochemical method and RT-PCR method detected of tumor cell cycle protein D (Cyclin D) expression.Result:Compare125I seeds experimental group with the control group, tumor volume significantly reduced, weight loss, each experimental group tumor volume growth curve decreased with time extension, and contrasted starkly with the control group increased. On the14day, The tumor volume inhibition rates of each experimental group were19%(B group),30%(C group),35%(D group), Compared the average volume between the experimental groups and the control group, the differences were statistically significant (P<0.05), Compared between the experimental groups, there were no significant differences (P>0.05);On the28day,The tumor volume inhibition rates of each experimental group are65%(B group),72%(C group),86%(D group), Compared between the experimental groups and the control group in the average volume, the difference is statistics significance (P<0.05), Compared between various experimental groups:D group with B, C two group the difference between groups is statistically significant (P<0.05), B group and C group the difference between groups is not statistically significant (P>0.05). Using TUNEL method to detect apoptosis display:Tumor cells in the control group grow good, apoptosis rate is low, Each experimental group can be visible more apoptosis cell, apoptosis index is increased; With the time prolonged, D group (0.8mCi) is obvious. Immunohistochemical detection of Cyclin D display:in14days, the experimental group Cyclin D protein expression were decreased, compared with the control group, the difference has statistical significance (P<0.05), the experimental group in Cyclin D protein expression has no significant difference (P<0.05);in28days, the experimental group Cyclin D protein expression were decreased, With the radioactivity increased Cyclin D protein expression was decreased, compared with the control group, the difference has statistical significance (P<0.05), the experimental group in Cyclin D protein expression has significant difference (P<0.05). RT-PCR for detection of Cyclin D shows:in Fourteenth days and twenty-eighth days, the experimental group in Cyclin D mRNA expression are reduced, and with the dose of radiation to increase Cyclin D mRNA expression is decreased significantly, compared with the control group, the difference was statistically significant (P<0.05); compared of each experimental group in Cyclin D mRNA expression, the difference was statistically significant (P<0.05). Compared respectively with two time point in each experimental group:group B (0.4mCi group) was not statistically different (P<0.05); and the C group (0.6mCi group) and group D (0.8mCi group) the difference has statistical significance (P<0.05).Conclusion:1.125I seed in organizational implantation for the treatment of human poorly differentiated gastric carcinoma is effective, can make human poorly differentiated gastric carcinoma transplanted subcutaneously in nude mice, tumor volume to reduct and weight to drop;2.125I seed can inhibit human poorly differentiated gastric cancer cell Cyclin D gene and protein expression, make tumor cell proliferation to reduce;3. Higher radiation activity of125I seed continuous irradiation induced apoptosis of human poorly differentiated gastric cancer cells (BGC823) apoptosis to increase more Obviously, we think that the single125I seed suitable activity range is0.6-0.8mCi. |
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