| Objective:(1) Grown in New Zealand white rabbits maxillofacial the VX2tumor lines, and combined with CT findings and pathology biopsy, the establishment of animal models of head and neck cancer.(2) New Zealand white rabbits maxillofacial tumor cells, telomerase protein catalytic subunit (hTERT) activity, to explore the biological characteristics of the cell cycle indicators in tumor tissue.Methods:(1) Experimental groups:the experimental divided into2groups of30New Zealand white rabbits in group A as the experimental group of15New Zealand white rabbits were numbered Group A:T1, T2---T14, T15;group was the control group, a total of15New Zealand white rabbits were numbered group B:T1, T2---T14, T15; and each mark.(2) Test Method:rabbit VX2tumor strain cell suspension was inoculated into the left side of the group A rabbit maxillofacial masseter, the establishment of the left maxillofacial animal tumor models, saline injected into the left side of the group B rabbit Maxillofacial Ministry of masseter muscle as a control group. Using the TRAP assay (Telomeric Repeat Amplification Protocol) of rabbit tumor tissue cells telomerase protein catalytic subunit (hTERT) activity. Positive results on gel electrophoresis separated by6bp ladder, the number of bands and shades the size of the telomerase activity. Characteristics of the intracellular DNA and the binding of the fluorescent dye (such as propidium iodide-PI), the expression of cellular DNA content to detect rabbit maxillofacial tumor cell cycle index (SPF the PI and5cEx) expression.(3) Statistical analysis:In this study, using SPSS13.0for statistical analysis, experimental data are expressed as mean±standard deviation, the group see more analysis of variance (F test), the homogeneity of variance t test was used for pairwise comparisons, heterogeneity of variance is used t’test was used for pairwise comparisons, P<0.05as significant difference.Results:(1) Head and neck cancer animal model:VX2tumor cell suspension the rabbit group A and saline injected rabbits the fifth week of the B group were randomly selected, the left side of the line to the First Affiliated Hospital of Yangtze University maxillofacial CT scan the masseter seen in the left side of the A group mass, clear boundary, B group no mass generation. In inoculated from the5th week of random excision of the left side of the A group and B group maxillofacial pathological biopsy shows Group A:Remove the tumor tissues were sliced hematoxylin-eosin staining, pathology confirmed squamous cell carcinoma maxillofacial tumor tissue and muscle fiber junction, the muscle fibers of tumor invasion and maxillofacial tumor tissue, which is visible pathological mitotic; maxillofacial group B for the left side of the normal masseter muscle fibers.(2) TRAP-PCR method rabbit tumor cells, telomerase protein catalytic subunit (hTERT) activity:telomerase protein catalytic subunit (hTERT) expression in the electrophoresis gel presents a6bpd the ladder with map telomerase negative samples does not appear on this map, the tumor cells showed positive telomerase activity, three with the strongest performance of the experimental group A group of T2cells telomerase activity, and strip the most clear in the experiment was used as a positive control group, with a molecular weight marker DNA Markers fragment shows clear, with8as a negative control for the lysis buffer control no template negative control group, the experimental by TRAP assay experimental group A group of15patients with maxillofacial squamous cell carcinoma, detected expression of hTERT activity was positive in13cases, the positive rate was86.6%. Group B,15patients with maxillofacial hTERT activity was detected in the normal masseter muscle tissue positive expression for the two cases, the positive rate was13.3%. The experiments showed that in maxillofacial malignant tissue telomerase protein catalytic subunit (hTERT) activity was significantly higher than that in normal tissues (P<0.01), which can be drawn to the quantitative detection of telomerase activity can be distinguished Maxillofacial malignant tumor and normal tissue, this study further confirmed the relevance of telomerase activity expression and maxillofacial malignancy.(3) Telomerase protein catalytic subunit (hTERT) in rabbits maxillofacial tumor tissue detection of the cell cycle indicators:the animal model A group of15cases of squamous cell carcinoma of the maxillofacial B group (n=15) the normal masseter organizations were not in aneuploidy, and SPI, PI and5cEx the rabbit A group of maxillofacial malignant group mean:14.4%,19.58%and1.58%, respectively, compared to rabbits in group B normal maxillofacial masseter muscle group mean:3.33%,4.13%and0.02%, there was a clear increased, significant difference. SPF and the PI two sets of cell cycle index prompted Group A maxillofacial malignant parts of a high degree of cell proliferation, with the normal group B maxillofacial masseter muscle tissue significantly different, indicating that the maxillofacial malignant aneuploid compared to normal tissue diploid (P<0.01) differences5cEx malignant tumor tissue and normal tissue was significantly (P<0.05), and PI SPF in diploid tumors, aneuploid tumor and normal masseter muscle tissue compared to higher (P<0.01).Conclusions:(1) Established by taking the injection of VX2tumor cell suspension of rabbit and maxillofacial malignant tumor model has a high rate of tumor formation, tumor growth, cyclical, high stability, easy to operate.(2) In the rabbit maxillofacial malignant tissue protein telomerase catalytic subunit (hTERT) activity was significantly higher than normal tissue, telomerase activity maxillofacial malignant quantitative detection can be used as a diagnostic indicator of.(3) In the rabbit maxillofacial malignant tissue Gl/S phase cell cycle can be detected expression of telomerase protein catalytic subunit (hTERT) activity S of expression of hTERT activity was the most significant G2/M midterm barely detectable The hTERT no activity expression. |
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