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目的探讨骨形成蛋白9(BMP9)诱导C3H10T1/2细胞向心肌细胞样细胞分化过程β联蛋白(β-catenin)的作用。方法利用重组腺病毒BMP9(Ad-BMP9)诱导C3H10T1/2细胞分化,通过Western blot法检测Ad-BMP9及不同剂量XAV-939处理后β-catenin的激活情况;在完全抑制β-catenin的情况下,Ad-BMP9诱导1周,通过实时定量PCR法检测心肌细胞增强因子2C(MEF2C)、心肌组织特异性转录因子GATA结合蛋白4(GATA4);Ad-BMP9处理3周后,通过Western blot法及免疫荧光技术检测心肌连接子蛋白43(Cx43)、心肌肌钙蛋白T(c Tn T)的表达。结果在感染效率约50%的情况下,BMP9可过度激活β-catenin,使其磷酸化水平增高;在XAV-939抑制β-catenin后,Ad-BMP9诱导的C3H10T1/2细胞表达的c Tn T、GATA4、Cx43和MEF2C均受到抑制。结论β-catenin能被BMP9激活并参与BMP9诱导的C3H10T1/2细胞向心肌细胞样细胞分化。
Objective To investigate the effect of bone morphogenetic protein-9 (BMP9) on the differentiation of C3H10T1 / 2 cells into cardiomyocyte-like cells in the process of β-catenin. Methods The C3H10T1 / 2 cells were induced by recombinant adenovirus BMP9 (Ad-BMP9), and the activation of β-catenin after Ad-BMP9 and different doses of XAV-939 were detected by Western blot. When β-catenin was completely inhibited , And Ad-BMP9 for 1 week. Cardiomyocyte augmentation factor 2C (MEF2C) and cardiac tissue-specific transcription factor GATA4 (GATA4) were detected by real-time quantitative PCR. Immunofluorescence was used to detect the expression of cardiac connexin 43 (Cx43) and cardiac troponin T (cTn T). Results BMP9 overexpressed β-catenin and increased the phosphorylation of p-catenin at the infection efficiency of about 50%. After X-AR-939 inhibited β-catenin, the expression of cTn T in C3H10T1 / 2 cells induced by Ad-BMP9 , GATA4, Cx43 and MEF2C were inhibited. Conclusion β-catenin can be activated by BMP9 and is involved in the differentiation of C3H10T1 / 2 cells induced by BMP9 into cardiomyocyte-like cells.