骨形成蛋白Ⅱ型突变受体基因转染对口腔鳞癌细胞增殖和凋亡的影响

Effects of mutatant bone morphogenetic protein receptor-Ⅱ on the proliferation and apoptosis of oral squamous cell carcinoma cell line Tca8113

目的探讨骨形成蛋白(BMPs)与口腔鳞癌的发生、发展的可能关系。方法将含有BMP-Ⅱ突变受体的真核表达载体转染入Tca8113舌癌细胞,筛选和鉴定后,构建稳定表达BMP-Ⅱ突变受体的细胞株tBRII-Tca8113。对tBRII-Tca8113细胞和Tca8113细胞分别进行MTT检测,流式细胞仪(FCM)分析、BrdU标记检测细胞的增殖活性及DNA合成;检测tBRII-Tca8113细胞和Tca8113细胞的凋亡及细胞周期相关因子(CyclinD1,CDK-4,p27,p57)的表达。结果 Tca8113细胞和tBRII-Tca8113细胞的增殖指数MTT检测为0.47±0.01和0.35±0.008(t=22.953,P=0.000),BrdU检测为12.0±3.4和23.0±1.9(t=6.918,P=0.000),FCM检测为6.3和7.9;两组的凋亡指数为3.7±1.2和8.7±1.6(t=29.583,P=0.000);细胞周期因子在Tca8113和tBRⅡ-ca8113细胞中的平均灰度测量值为CyclinD1(186.5±2.4和145.6±3.9,t=28.244,P=0.000),CDK4(169.9±2.9和129.5±3.2,t=29.583,P=0.000),p2(7110.1±1.1和167.3±1.8,t=85.754,P=0.000),p5(7107.9±2.1和156.8±2.2,t=50.844,P=0.000)。结论 BMPs及其受体可能在口腔上皮组织的恶变过程中有重要作用,研究结果为探讨BMPs信号在上皮性肿瘤中的作用提供了重要依据。tBRII-Tca8113细胞的建立,进一步表明了BMPs及其受体在口腔上皮组织的恶变过程中有重要作用,并为进一步探讨BMP信号在上皮性肿瘤的恶变过程中的作用奠定了基础。

Objective To investigate the possible roles of Bone Morphogeneti c Proteins （BMPs） on the occurrence of oral squamous cell carcinoma. Methods A negative truncated mutant of the BMP receptor - Ⅱ （tBMPR- Ⅱ ） was transfected into Tea8113 cells to obtain tBRII- Tea8113 cell line which expressed BMP-Ⅱ mutant stably. Cell activities and DNA synthesis in tBRII-Tca8113 cells and Tca8113 groups were evaluated though MTT assay, Flow cytometry （FCM） analysis and BrdU staining. TUNEL staining, and cell cycle related proteins （CyclinD1, CDK-4, p27, p57） were also detected to observe the changes of apoptosis and cell cycle. Results Values of proliferation index, BrdU index, FCM, apoptosis index and average gray of cell cycle related proteins were 0.47 ± 0.01and 0.35 ±0.008, BrdU index were 12± 3.4 and 23 ± 1.9, 6.3 and 7.9, 3.7 ± 1.2 and 8.7±1.6, Cyclin D1 （186.5 ± 2.4, 145.6 ± 3.9）, CDK4 （169.9 ± 2.9, 129.5 ± 3.2）, p27 （110.1± 1.1, 167.3 ± 1.8）, p57 （107. 9 ± 2.1, 156.8 ± 2.2）, respectively in Tca8113 and tBRⅡ-Tca8113 groups. Significant differences were found in all of these databetween two groups.Conclusion BMPs and its receptors play critical roles in the malignant transformation of oral epithelium. Our results provide important evidences for the BMPs attributing to the formation of epithelial tumors. The establishment of tBR Ⅱ-TcaS113 cell line laid the solid foundation for the further research of BMPs effects on the epithelial tumors.