Growth Suppression On The Nude Mice Model Of Esophagus TE13Cell Carcinoma By Introduction Of RIZ1Gene

[Objective]Through the establishment of human esophageal TE13cells in nude mice transplantation tumor models,inject RIZ1eukaryotic expression plasmid of esophageal TE13cells in nude mice transplantation tumor, explores the inhibitory effect of RIZ1eukaryotic expression plasmid of esophageal TE13cells in nude mice transplantation.[Methods]1. Cell cultureConventional methods recover esophagus carcinoma TE13cell lines,in DMEM what containing10%fetal bovine serum regular batches. Cell culture time and quantity meet the experimental requirements,charge for cell and cell count.2. Establish the esophageal TE13cells nude mice transplantation tumor model and the experiment groupingIn the fourth day, we collect TE13cells which in logarithmic phase, and inject the liquid of cell in each nude mice right armpit. The grouping is devided as follows:(1):the blank control group, injected NS100μL each time;(2) empty negative plasmid group:liposomes20μL+Empty plasmid20μg, add the DMEM medium liquid quantity to100μL each time;(3)pcDNA3.1(+)/RIZ1experimental group: liposome20μL+RIZ1eukaryotic expression plasmid20μg, add the DMEM medium liquid quantity to100μL each time; Three groups all use the method of tumor multi-point injection, every other two days, a total of5times.3. To detect RIZ1mRNA and its related protein expression using the method of RT-PCR, Western blot technique4. The morphology observation of human esophageal TE13cells in nude mice transplantation tumorObserve the nude mice general situation and transplantation tumor growth change; measure the length and the width of the tumor; Calculate the tumor volume; Draw the tumor growth curve. HE staining.[Results] 1. Succeed to build a esophageal TE13cells in nude mice transplantation tumor model. It has the feature of tumor cells showed HE staining results.2. The expression of RIZ1gene of human esophageal cancer TE13cells in nude mice transplantation tumorAfter doing the RT-PCR of the RNA extracting from the transplantation tumor, the product electrophoresis results showed that:three sets of the endogenous GAPDH bands with similar strength brightness, while pcDNA3.1(+)/RIZ1group appears clear bands between100bp-200bp, its size is consistent with the theory of expected value (167bp). Using Western blot detect RIZ1of each group, the results showed that: pcDNA3.1(+)/RIZ1can be seen obviously RIZ1expression, while the blank control group and the empty plasmid group have lower expression of RIZ1.3. The inhibition effect of the RIZ1eukaryotic expression plasmid of esophageal TE13cells to the nude mice transplantation tumor growthDispose these groups of nude mice in the7th day after the injection of RIZ1eukaryotic expression plasmid.Experimental group tumor volume and weight are much smaller than the other two groups, the difference is statistically significant (P<0.05); The blank control group comparing with empty plasmid negative control group there was no statistically significant difference (P>0.05). The ratio of inhibiting tumor is53.6%.[Conclusion]1. Human esophageal carcinoma TE13cells can be applied to establishment of transplantable model of human esophageal carcinoma in nude mice2. Using liposome mediated method of RIZ1eukaryotic expression plasmid which can realize the purpose gene in esophageal TE13stable expression in nude mice transplanted tumor cells3. The tumor injection of RIZ1eukaryotic plasmid expression has inhibition effect to the esophageal TE13cells in nude mice transplantation tumor...