The Study Of The Effect And Mechanism Of Bone Morphogenetic Proteins-2 On The Tca8113 Cell Of Tongue Cancer

Bone morphogenetic proteins-2 was found firstly as an important factor in regulating bone formation, it plays an important role in embryonic skeletal development and regeneration. Now studies suggest that, it is involved in the regulation of many kinds of cell proliferation, differentiation and apoptosis of biological process. A growing number of studies have shown that BMPs signaling pathways involved in tumor progression. This study aims to transfect adenovirus-mediated BMP2 into the tongue cancer cell Tca8113, and then observe the changes of biological behavior of tongue cancer cell line Tca8113 after transfected with BMP2, and to detect the change of BMP-SMAD pathway before and after transfection, explore the regulatory mechanism of molecular biology of BMP2 to tongue cancer, in order to seek a more effective method of gene therapy.Objective:To observe the effect of BMP2 on the biological behavior of tongue cancer cells, and study the regulation of BMP-SMAD pathway on tongue cancer cells, establish the animal model of tongue cancer, and observe the effect of BMP2 on the nude with tongue cancer.Methods:1. Cultivation of tongue cancer cell:recovery tongue cancer cell line Tca8113, cultivated in RPMI1640 in vitro, and observe the cell morphology and proliferation activity.2. Observe the effects and mechanism of BMP2 on Tca8113 cell:use different density of Ad-BMP2 to transfect Tca8113 cell, and detect the expression of BMP2, Smad1/5 by Western blot.3. Observe the effect of BMP2 on Tca8113 cells biological behavior:transfect enhanced green fluorescent BMP2 gene to Tca8113 cells by adenovirus,observe the changes in cell morphology,detect the changes in cell proliferation activity, and detect the pathways of Smad.4. Experiment in vivo:establish tumor models in SCID mice inguinal subcutaneous, inject adenovirus-mediated BMP2 gene to the tumor after the formation of the models, and observe the changes in tumor volume.Results:1. Cultivation of tongue cancer density cell:Tca8113 cells grow stably and have normal morphology.2. After the transfection of adenovirus-mediated BMP2 gene to Tca8113 cells, Western blot detected significant different expression of BMP2 and Smadl/5 among different groups,the higher density of transfection group BMP2, the higher expression levels of Smadl/5.3. After the transfection of adenovirus-mediated enhanced green fluorescent BMP2 gene to Tca8113 cells, cellular morphology do not change. MTT detects that cell proliferation activity was significantly lower, and fluorescence expressed clearly in the nucleus.4. Tumors on the SCID mice subcutaneous grow to an average of about 0.6 cm in diameter after 2 weeks. Adenovirus-mediated BMP2 gene is injected to the tumor, and the tumor was suppressed. The tumor size is different among the groups.Conclusions:1. Adenovirus-mediated BMP2 genes are transfected to Tca8113 cells successfully, and they were identified by Western blot. The interest protein has been integrated into Tca8113 cells with effective expression.2. After transfecting BMP2 to Tca8113 cells, the expression of Smadl and Smad5 protein significantly increase.3. BMP2 inhibites the proliferation of Tca8113 obviously, and the signal of BMP2 is transducted from extracellular to nucleus by Smad1/5 proteins, and then BMP2 plays regulatory role in the nucleus.4. There is no significant effect on the morphology of the tongue cell Tca8113 of BMP2.5. Tongue animal model is successfully constructed, and BMP2 inhibites the growth of transplantable tumor in SCID mice.