| Tongue cancer, which accounts for45.9%of the oral and maxillofacial maligancies, is one of the most common malignant tumors in the oral and maxillofacial region.The The etiology of tongue cancer remains unknown, the theory of" combined effects of carcinoid tumor causes" were generally accepted. Therefore, it is important to investigate the development mechanisms and treatment of tongue cancer. With the recent progresses in molecular biology field, researchers believe that the occurrence of tongue cancer is a process comprised of more than one gene and many cytokines. Bone morphogenetic protein (BMP) and its signaling pathway were found to play an important role in the occurrence and development of tongue cancer in recent studies.Bone morphogenetic protein2(Bone morphogenetic protein-2, BMP-2) is an important factor in regulating bone formation, which plays an important role in embryonic development and regeneration of bone repair. Recent studies have found that BMP had an impact on the proliferation, formation and metastasis of different malignant cells and participated in regulating the malignant biological behavior. Noggin gene plays an important role in the regulation of the development and/or remodeling of multiple skeletal system and nervous system in the body. Noggin protein is one kind of BMP-2extracellular antagonists, it inhibits the function of BMPs through specifically combining with BMPs and preventing the binding of BMPs and cell surface receptor. Noggin gene plays an important role in the regulation of the development and/or remodeling of multiple skeletal system and nervous system in the body. Noggin protein is one kind of BMP-2extracellular antagonists, it inhibits the function of BMPs through specifically combining with BMPs and preventing the binding of BMPs and cell surface receptor.Objcetive:To observe the effects of BMP-2and the interaction between BMP-2and Noggin on the proliferation of tongue cancer cells, as well as the changes of the BMP/Smad signal transduction pathway.Methods:1. The culture of tongue cancer cell:Tca8113cells were cultured in vitro and the morphology and proliferative activity of cells were observed.2. Different concentrations of BMP-2protein (0,20,40,60,80,100ng/ml) were applied to the tongue cancer cells cultured in vitro. The proliferation and morphology of cell were observed under inverted microscope. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the proliferation of tongue cancer cells co-cultured withdifferent concentrations of BMP-2at day1,2,3,4,5. Western blot was used to detect the expression of BMPR â…¡ and Smad4at different BMP-2concentrations (40,80ng/ml) at the second and fourth day.3. Cells were cultured with medium containing40ng/ml BMP-2, Noggin protein was added to the medium with concentration of0,100,200,300,400,500ng/ml respectively after24h. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the effects of Noggin and BMP-2on the proliferation of tongue cancer cells at day1,2,3,4,5. Western blot was used to detect the expression of BMPR â…¡ and Smad4at different BMP-2concentrations (200,400ng/ml) at the second and fourth day.Results:1. Tongue cancer Tca8113cell grew well. The single cell was in the form of triangle or rectangle, with less stereoscopic effect. They lived in groups, proliferate quickly, enter logarithmic growth phase at the second day, and flat phase at the fourth day.2. BMP-2inhibited the proliferation of tongue cancer cell, and the inhibition effect was concentration-dependent. A strong inhibition was observed at concentration of80ng/ml, while a stable inhibition was seen at concentration of100ng/ml. The expression of BMPR â…¡ and Smad4increased with the concentration of BMP-2and time.3. Noggin promoted the proliferation of tongue cancer cell, and the promotion was concentration-dependent. The most effective concentration was400ng/ml, while a stable promotion was seen at concentration of500ng/ml.The expression of BMPR â…¡ and Smad4decreased with the concentration of Noggin and time.Conclusion: The MTT results show that exogenous BMP-2protein inhibits the proliferation of tongue cancer cells with time-dependent and dose-dependent. And the Western Blot results display that the stronger BMP-2protein inhibits the proliferation of tongue cancer cells, the stronger expression of BMP/Smad signal transduction pathway is, which indicates that BMP-2protein might inhibits the proliferation of tongue cancer cells by BMP/Smad signal transduction pathway.The MTT results show that the Noggin protein as BMP-2protein antagonist promotes proliferation of tongue cancer cells by blocking the inhibition effect of BMP-2on proliferation of tongue cancer cells. And the Western Blot results display that the higher concentration of Noggin protein is, the weaker expression of BMP/Smad signal transduction pathway is, which further evidence that the inhibition effect of BMP-2protein on proliferation of tongue cancer cells may be associated with the BMP/Smad signal transduction pathway. |
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