Suppression to ameloblastoma xenografts of chicken embryo chorioallantoic membrane by tissue inhibitor of metalloproteinases-2 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore the invasiveness of xenografts on chicken embryo chorioallantoic membrane (CAM) after tissue inhibitor of metalloproteinase-2 (TIMP-2) gene transfection.</p><p><b>METHODS</b>Fresh ameloblastoma tissues were minced into 1-2 mm3 and transplanted on the CAM. There were three groups named as control group (Empt), plasma transfection group (Lipo), and TIMP-2 gene transfection group (P). The specimens were respectively investigated by microscope indifferent spots after implanting. The volume of the xenografts and the weight of xenografts in the termination time of the experiment were recorded. The invasiveness of xenografts was divided into four grades by pathological examination. Western blot analysis was performed to investigate matrix metalloproteinase-2 (MIMP-2) and TIMP-2 protein in xenografts.</p><p><b>RESULTS</b>Ameloblastoma tissues can survive on CAM and the tumor cells may invade it on 5-7 days after implanting. At 9 d after implanting, the invasiveness grades in P group were 7 in grade 0, 1 in grade 2, 0 in grade 3. The expression of TIMP-2 protein in P group was significantly higher than that in Empt group (P < 0.05). The expression of MMP-2 protein in P group was lower than that in Empt group (P < 0.05).</p><p><b>CONCLUSION</b>The xenotransplanted tumor model of human ameloblastoma on CAM was successfully established. The invasiveness of ameloblastoma xenografts was suppressed might be due to TIMP-2 gene transfection.</p>

Establishment of subcutaneous xenotransplanted tumor model of human ameloblastoma in nude mice / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To establish an subcutaneous xenotransplanted tumor model of human ameloblastoma in nude mice.</p><p><b>METHODS</b>Ameloblastoma cells were absorbed by primary culture, repeat attachment and pancreas proteolytic enzyme were both used to purify them. Then, the purified cells were implanted subcutaneously into the nude mice. The specimens were respectively investigated by microscope in different spots after implanting.</p><p><b>RESULTS</b>Ameloblastoma cells can survive in all of the 8 nude mice. The xenograft can be found on 23 days after implanting. The rate of successful inocutation is 25%. The subcutaneously xenotransplanted tumor cells can be found with microscope in the inter-muscle tissues of nude mice.</p><p><b>CONCLUSION</b>The subcutaneously xenotransplanted tumor model of human ameloblastoma in nude mice was successfully established and it may benefit to further studies on this tumor.</p>

The construction and expression of PcDNA3.1(+)/GFP-TIMP-2 in human ameloblastoma cell / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector of TIMP-2 gene and to explore its expression in human ameloblastoma cell in vitro.</p><p><b>METHODS</b>The aimed gene fragment was obtained by RT-PCR. And then, molecmicrolar cloning technology and enzyme digestion were used to connect the gene with the plasmid PcDNA3.1(+), which can be expressed in eukaryotic cells and a report gene: green fluorescent protein gene (GFP) was already existed in the plasmid. We named the eukaryotic expression vector, which contended our aimed gene TIMP-2 as well as report gene GFP, PcDNA3.1(+)/GFP-TIMP-2. The vector was identified by PCR analysis, EcoR I and Xho I restriction analysis and Sequence analysis. After the PcDNA3.1(+)/GFP-TIMP-2 was transfected into cultured human ameloblastoma cell, RT-PCR and Flow Cytometry (FCM) and Microscope wre respectively performed to evaluate the effect of transfection and expression.</p><p><b>RESULTS</b>The constructed vector PcDNA3.1(+)/GFP-TIMP-2 was proved correct by enzyme digestion and sequencing analysis. After PcDNA3.1(+)/GFP-TIMP-2 was trasnfected into cultured human ameloblastoma cell, the rate of transfection is 47.6% (Analysis report of FCM), the green fluorescence was found in plasm (observed with fluo-microwave), the expression of TIMP-2 mRNA was elevated 2.4 times compared with the control group.</p><p><b>CONCLUSIONS</b>PcDNA3.1(+)/GFP-TIMP-2 was successfully constructed and it could be transfected into cultured human ameloblastoma cell. It may be benefit to further study of the relationship between the TIMP-2 gene and the behaviour of ameloblastoma.</p>