骨形态发生蛋白2基因转染对舌鳞癌细胞体内外生物学行为的影响

Effects of bone morphogenetic proteins-2 gene transfection on biologic behaviors of tongue cancer cells in vitro and in vivo

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 adenovirus-mediated BMP2 （Ad-BMP2） gene transfection to Tca8113 cells in vitro, and observe the effect of BMP2 gene transfection on the severe combined immunodeficiency （SCID） mice with tongue cancer line Tca8113 in vivo. Methods Tongue cancer cell line TcaS113 was culti- vated in vitro and transfected with different densities of Ad-BMP2. Expressions of BMP2, Smadl and Smad5 in the Ad- BMP2 and PBS groups were detected through Western blot. Tumor models in SCID mice inguinal subcutaneous were established and injected with Ad-BMP2 gene. The changes of tumor volume and transferring circumstances were observed. Results The result demonstrated that the expression levels of Smadl and Smad5 were positively correlated to the BMP2 transfection density（ P 〈 0.05 ）. After the Ad-BMP2 gene was transfected into Tca8113 cells, the tumors diameter of the SCID mice grew about 0.6 cm in 2 weeks, and the tumor growth was obviously inhibited. The metastases of tumor in the liver, spleen, kidney and tomentum majus of the SCID mice were not detected. Conclusion Ad-BMP2 gene is successfully transfected to Tca8113 cells and can significantly increase the expressions of Smadl and Smad5 proteins. BMP2 can significantly inhibit the growth of Tca8113 tumor grafted on the SCID mice.