论文部分内容阅读
目的检测脊髓性肌萎缩症(SMA)患者神经元样细胞 SMN2基因 mRNA 的表达。方法用 PCR-RFLP 的方法对 SMA 患者进行基因诊断,诱导其骨髓间充质干细胞分化为神经元样细胞,RT-PCR 和测序检测该细胞 SMN 2基因 mRNA 的表达,所有过程与对照组进行对比研究。结果SMN2基因的扩增产物为全长转录产物 fl-SMN mRNA(266 bp)和转录时跳过外显子7的产物 SMN△7mRNA(212 bp,经测序证实缺少的54 bp 为外显子7的序列);SMA 患者 fl-SMN mRNA 的表达占总表达量的23.2%,远低于对照者(占总表达量的82.0%);而 SMN△7 mRNA 表达占总表达量的76.8%,高于对照者的18.0%。结论 SMA 患者神经元样细胞的 SMN2基因转录时存在选择性剪接,即剪接时跳过外显子7,是导致其全长 SMN 蛋白不足的原因之一。
Objective To detect the expression of SMN2 mRNA in neuron - like cells of spinal muscular atrophy (SMA) patients. Methods The gene of SMA patients was diagnosed by PCR-RFLP. The bone marrow mesenchymal stem cells were induced to differentiate into neuron-like cells. The expression of SMN 2 gene mRNA was detected by RT-PCR and sequencing. All the process was compared with the control group the study. Results The amplified product of SMN2 gene was the full-length transcript fl-SMN mRNA (266 bp) and SMNΔ7 mRNA (212 bp), a product of skipping exon 7 when transcribed. The missing 54 bp was exon 7 ). The expression of fl-SMN mRNA in SMA patients was 23.2% of the total, which was much lower than that in controls (82.0% of the total), while the expression of SMN △ 7 mRNA was 76.8% of the total 18.0% of the controls. Conclusion The splicing of SMN2 gene in neuron-like cells of SMA patients during splicing, that is, skipping of exon 7 during splicing is one of the reasons for the lack of full-length SMN protein.