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目的探讨骨形态发生蛋白2(BMP2)基因转染对体外舌鳞癌细胞的作用及机制,并观察体内转染BMP2基因对严重联合免疫缺陷(SCID)小鼠皮下移植瘤生长与转移的影响。方法体外培养舌癌Tca8113细胞,将重组腺病毒介导的BMP2(Ad-BMP2)基因分别按0、50、100感染复数(MOI)转染Tca8113细胞后,采用Western blot检测Ad-BMP2组和PBS组细胞中BMP2、Smad1和Smad5表达的差异。建立SCID鼠肿瘤模型并瘤内注射Ad-BMP2基因,观察肿瘤体积的变化及肿瘤转移情况。结果 Ad-BMP2基因成功转染至Tca8113细胞,MOI为100时转染效率较高,Ad-BMP2组和PBS组细胞中BMP2、Smad1和Smad5的表达差异有统计学意义,P<0.05,Ad-BMP2组Smad1和Smad5的表达量与BMP2的浓度呈正比。SCID鼠皮下移植瘤的体积在2周后平均增大0.6 cm3,瘤内注射Ad-BMP2基因后,明显抑制移植瘤的生长。肝、脾、肾、大网膜等脏器均未查见肿瘤转移。结论以腺病毒为载体的BMP2在Tca8113细胞中有效表达,Smad1和Smad5蛋白的表达也显著提高。基因转染BMP2显著抑制SCID鼠舌癌移植瘤的生长。
Objective To investigate the effect and mechanism of BMP2 gene transfection on human tongue squamous cell carcinoma in vitro and to observe the effect of transfection of BMP2 gene in vivo on the growth and metastasis of subcutaneously transplanted tumor in mice with severe combined immunodeficiency (SCID). Methods Tca8113 cells were cultured in vitro. Tca8113 cells were transfected with recombinant adenovirus-mediated BMP2 (Ad-BMP2) gene at the MOI of 0, 50 and 100, respectively. Differences in expression of BMP2, Smad1 and Smad5 in group cells. Establishment of SCID murine tumor model and intratumoral injection of Ad-BMP2 gene, observe the tumor volume changes and tumor metastasis. Results Ad-BMP2 gene was successfully transfected into Tca8113 cells. The transfection efficiency was higher at MOI 100, and there was significant difference in the expression of BMP2, Smad1 and Smad5 in Ad-BMP2 and PBS groups (P <0.05, Ad- The expression of Smad1 and Smad5 in BMP2 group was directly proportional to the concentration of BMP2. The volume of subcutaneously transplanted tumors in SCID mice increased by an average of 0.6 cm3 after 2 weeks. After intratumoral injection of Ad-BMP2 gene, the growth of transplanted tumors was obviously inhibited. Liver, spleen, kidney, omentum and other organs were not found in tumor metastasis. Conclusion Adenovirus-mediated BMP2 is efficiently expressed in Tca8113 cells, and the expression of Smad1 and Smad5 proteins is also significantly increased. Gene transfection BMP2 significantly inhibited the growth of SCID mouse tongue cancer xenografts.