A Study On The Influence Of Proliferation And Apoptosis For Tum-5 Gene To Human Gastric Carcinoma Cell

This study based on gastric cancer cells(SGC-7901). PcDNA3.1-Tum-5 eukaryotic plasmid carrier is constructed successfully by using nested PCR technology. After that it was transfected with lipofectamine to stomach cancer line, meanwhile,to detect the expression of Tum-5 in gastric cancer cells .To observe the change of gastric cancer cell apoptosis, growth cycle and cell proliferation;.The objective is to explore a new anti-tumor angiogenesis inhibitors Tumstatin and provide the evidence of solid tumor therapy through the experiment research. Objective:To construct the recombinant plasmid carrier pcDNA3.1-Tum-5 and was transfected with lipofectamine to stomach cancer line. And to detect t the change of gastric cancer cell apoptosis, growth cycle and cell proliferation of gastric cancer cells .then to explore the influence of anti-tumor angiogenesis inhibitors Tumstatin.Methods:1 PcDNA3.1-Tum-5 eukaryotic plasmid carrier is constructed by using nested PCR technology.2 The recombinant plasmid carrier pcDNA 3.1-Tum-5 was transfected with lipofecta- mine2000 to human stomach cancer SGC-7901(experiment group), blank plasmid carrier as comparison in this experiment. and measured the transfection efficiency with fluorescence microscope .meanwhile, to detect the expression of mRNA Tum-5 with RT-PCR in tomach cancer line in three groups.3 The method of MTT was used to measure the proliferation inhibition rates of stoma- ch cancer cells after incubated 24 hours ,48 hours and 72 hours respectively.4 To observe gastric cancer cell apoptosis, growth cycle in flow cytometry. 1 PcDNA3.1-Tum-5 eukaryotic plasmid carrier is constructed successfully by using nested PCR technology.2 The recombinant plasmid carrier pCDNA3.1-Tum-5 was transfected with lipofectamine in vitro was failed to stomach cancer line stably expressed,but transient transfection was succeed and fluorescence microscope measure its transfection efficiency for 90 ~ 95%. The recombinant plasmid carrier can normal express in RT - PCR in gastric cancer cells.3. The results of MTT Experiment discovered that the proliferation inhibition rates of experimental group were significantly higher in control group and the negative control group (P < 0.05), including each cell proliferation 72h after carrying 48h, inhibiting rates are higher than 24h (P < 0.05) .4 The results of gastric cancer cell apoptosis in flow cytometry analysis after transfected 48 hours showed that: the apoptosis cells percentage of experimental group without serum 6 hours and 12 hours were higher than the corresponding control group .The cell cycle testing results show that the G1 phase, G2 period, S period percentage did not change significantly, and G2 / G1 scale also did not change significantly .Conclusion: PcDNA3.1-Tum-5 eukaryotic plasmid carrier is constructed success -fully by using nested PCR technology. And the recombinant plasmid carrier can normal express after incubated in vitro in gastric cancer cell SGC-7901. Tum-5 genes And can significantly inhibited gastric cancer cell increment, promote cell apoptosis, but on the growth cycle without obvious influence.