目的探讨基质金属蛋白酶组织抑制剂-2(TIMP-2)基因过表达对人成釉细胞瘤鸡胚尿囊膜(CAM)移植瘤的抑制作用。方法建立成釉细胞瘤的鸡胚尿囊膜移植瘤模型。将实验分组:空白对照组(Empt),脂质体转染组(Lipo),质粒转染组(P)。TIMP-2基因转染成釉细胞CAM移植瘤细胞后,测定移植瘤体积和瘤重;将移植瘤侵袭能力分为4个级别进行移植瘤病理检查。Western blot分析移植瘤基质金属蛋白酶-2(MMP-2)及TIMP-2蛋白的表达。结果移植瘤能在鸡胚绒毛尿囊膜上生长。接种后第9天,转染质粒组的0级侵袭为7例、2级侵袭为1例、3级侵袭为0例。TIMP-2基因转染可以显著抑制成釉细胞CAM移植瘤的局部侵袭能力。质粒转染组TIMP-2表达明显高于空白组TIMP-2的表达(P〈0.05);质粒转染组MMP-2表达明显低于空白组MMP-2的表达(P〈0.05)。结论成功建立成釉细胞瘤的CAM移植瘤模型。TIMP-2基因转染后,成釉细胞瘤CAM移植瘤的侵袭性生长被抑制。移植瘤的侵袭性生长被抑制的可能原因是由于特异性抑制MMP-2蛋白引起的。
Objective To explore the invasiveness of xenografts on chicken embryo chorioallantoic membrane(CAM) after tissue inhibitor of metalloproteinase-2(T IMP-2)gene transfection.Methods 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 in different 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(MMP-2) and TIMP-2 protein in xenografts.Results 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(P0.05).The expression of MMP-2 protein in P group was lower than that in Empt group(P0.05).Conclusion 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.