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目的观察LYRM1基因过表达对P19细胞凋亡的影响。方法将人LYRM1基因的真核表达载体(pcDNATM3.1/myc-HisB-LYRM1)和空载体pcDNATM3.1/myc-HisB,分别转染P19细胞,通过G418筛选2周,建立稳定过表达人LYRM1基因的P19细胞系。用Western blot技术验证稳定表达细胞株。将稳定转染pcDNATM3.1/myc-HisB-LYRM1载体质粒和空载质粒的P19细胞以无血清培养基37℃孵育24h以诱导细胞凋亡,收获细胞用流式细胞术检测细胞凋亡。结果建立了稳定转染LYRM1的P19细胞系,成功地表达了目的基因。流式细胞术检测结果发现过表达LYRM1基因的P19细胞凋亡率显著降低。结论 LYRM1基因具有抑制P19细胞凋亡的作用。
Objective To observe the effect of LYRM1 gene overexpression on the apoptosis of P19 cells. Methods The human LYRM1 gene eukaryotic expression vector (pcDNATM3.1 / myc-HisB-LYRM1) and the empty vector pcDNATM3.1 / myc-HisB were transfected into P19 cells respectively and screened by G418 for 2 weeks to establish stable overexpression human LYRM1 Gene P19 cell line. The stable expression of cell lines was verified by Western blot. P19 cells stably transfected with pcDNATM3.1 / myc-HisB-LYRM1 vector and empty vector were incubated with serum-free medium at 37 ° C for 24 h to induce apoptosis. Cells were harvested for cell apoptosis assay by flow cytometry. Results The P19 cell line stably transfected with LYRM1 was established and the target gene was successfully expressed. Flow cytometry results showed that the apoptosis rate of P19 cells over-expressing LYRM1 gene was significantly reduced. Conclusion LYRM1 gene can inhibit the apoptosis of P19 cells.