| BMPs (Bone morphogenetic proteins, BMPs) belong to the superfamily of transforming growth factor-(3(TGF-β). In the early days, it caused for concern because the ability to stimulate bone cell differentiation; in recent years, it had been founded in the expression of tooth development related in original oral epithelium. The BMP family members could be founded in malignant tumors such as in tongue cancer cells, and they participate in the occurrence and development of tumor. BMP2(Bone morphogenetic proteins2, BMP2) is one of the most representative factors of BMPs family, besides its bone repair effect we also founded that it can inhibit the growth of epithelial cancer especially in tongue cancer cells, the signaling pathways of BMP have inseparable relationship with the growth progression of tumor. BMP signaling pathways were composed with BMP, BMP receptor complexes (BMP-receptor, BMPR) and SMAD, BMPR is an important part of BMP signaling pathway, it divided into BMPR-I (including BMPR-IA and BMR-IB) and BMPR-II. When the expressions of BMP receptor are restrainnig, they inhibit the differentiation of cancer cells through block the BMP signaling pathways at a certain extent.Tumor necrosis factor-alpha (TNF-a) plays a key role in the progression of host resistance to microbial invasion and tumor cell defense system, it involved in many biological and pathological activity processes. This study through observing the changes of biological behavior of tongue cancer cell in different concentrationes of TNF-a that treated with rhBMP2; Study the influence and mechanism of BMP-SMAD pathway in tongue cancer cell by detected the proteins of Smad4, BMPR-IA, BMPR-IB and BMPR-II, in order to explore the feasibility of TNF-a and rhBMP2on antitumor activity in vitro and growth inhibition of oral cancer in vivo.Objective:To observe the influence of activity proliferation of tongue cancer cell threated with TNF-a and rhBMP2, and study the influence and expression differences about of signal protein Smad4, BMPR-IA, BMPR-IB and BMPR-II, that are closely related to the progress of tongue cancer.Methods:1. To recover tongue cancer cell lines and observe the proliferat activity:Recover tongue cancer cell lines Tca8113and CAL27in RPMI1640high sugar culture medium in vitro, observe the morphology and proliferation activity of these cells. To detect proliferation activity of tongue cancer cells in different groups by MTT. Observation the behavior influence and the morphology change of tongue cancer cells biology in TNF-a groups and TNF-a+rhBMP2groups by RT-PCR.2.Detect the different expressiones of mRNA of BMPR-IA, BMPR-IB, BMPR-II and Smad4in TNF-a groups and TNF-a+rhBMP2groups by RT-PCR.3.Detect the different expressiones of proteins of BMPR-IA, BMPR-IB, BMPR-II and Smad4in TNF-a groups and TNF-a+rhBMP2groups by Western blot.Results:1. The growth of Tongue cancer cells of Tca8113and CAL27cell was stabilized, they had normal morphology and proliferated faster, they had been cultivated into the period of logarithmic growth and plateau at the48h and72h respectively.2.MTT results showed TNF-a can inhibit the growth of tongue cancer cells, especially when it had acting on tongue cancer cells treated with rhBMP2, the proliferation activity of tongue cancer was decreased significantly.3.Compared with the control group, RT-PCR detected that the mRNA expression of Smad4, BMPR-IA, BMPR-IB and BMPR-II were higher in the TNF-a group, and the mRNA expressions of Smad4, BMPR-IA, BMPR-IB and BMPR-II were the highest in the TNF-a+rhBMP2group.4.Western blot found the obvious difference of the protein expression of Smad4, BMPR-IA, BMPR-IB and BMPR-II in the different groups, compared with the control group, the TNF-a group has higher protein expression, and the TNF-a+rhBMP2group has the highest protein expression.Conclusion:1.The rhBMP2and TNF-a had no affect on the shape of tongue cancer cells. TNF-α can inhibited the proliferation of tongue cancer cells.2.The inhibition of cell proliferation was more significant in the TNF-α+rhBMP2group compared with the TNF-a group.3.The expressiones of proteins and mRNAs of Smad4, BMPR-IA, BMPR-IB and BMPR-II were increased obviously. |
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