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目的:检测BRCA1及其剪接异构体BRCA1△11b在P19及人胚胎干细胞分化过程中的表达。方法:全反式维甲酸诱导P19细胞的分化,提取不同时间点RNA,运用RT-PCR,Real-time PCR分别检测P19细胞及人胚胎干细胞分化过程中BRCA1及其剪接异构体BRCA1△11b的表达。结果:未分化P19细胞在全反式维甲酸诱导下聚集形成拟胚体。在P19细胞拟胚体形成过程中,BRCA1△11b表达水平迅速降低,而BRCA1的表达则随P19细胞分化发生相反的变化,BRCA1表达水平迅速升高。在人胚胎细胞分化过程中,BRCA1基因同样发生选择性剪接的转换,由未分化时BRCA1△11b为主要剪接产物转换为拟胚体形成时BRCA1表达占优势。结论:BRCA1及其选择性剪接异构体BRCA1△11b在P19细胞及人胚胎干细胞分化过程中均发生选择性剪接的转换,BRCA1基因的选择性剪接可能在胚胎干细胞的分化过程中发挥重要作用。
AIM: To detect the expression of BRCA1 and its splicing isoform BRCA1 △ 11b in P19 and human embryonic stem cells. Methods: All-trans retinoic acid induced differentiation of P19 cells and RNA extracted at different time points. RT-PCR and Real-time PCR were used to detect the expression of BRCA1 and its isoform BRCA1 △ 11b in human embryonic stem cells expression. Results: Undifferentiated P19 cells induced the induction of all-trans retinoic acid to form embryoid bodies. In the process of P19 embryoid body formation, the expression level of BRCA1 △ 11b decreased rapidly, while the expression of BRCA1 changed inversely with the differentiation of P19 cells, and the expression level of BRCA1 increased rapidly. In the process of human embryonic cell differentiation, the BRCA1 gene also undergoes alternative splicing transformation. The expression of BRCA1 predominates when the undifferentiated BRCA1 △ 11b is the major splicing product and the embryoid body is formed. CONCLUSION: BRCA1 and its alternative splicing isoform BRCA1 △ 11b undergo alternative splicing during the differentiation of P19 cells and human embryonic stem cells. The alternative splicing of BRCA1 gene may play an important role in the differentiation of embryonic stem cells.