细胞周期蛋白D2的RNA干扰质粒转染对骨髓瘤细胞系LP-1细胞增殖和凋亡的影响

目的构建细胞周期蛋白 D2(CCND2)的短发夹 RNA 表达质粒,并观察对骨髓瘤细胞系 LP-1细胞增殖和凋亡的影响:方法设计、合成 CCND2短发夹 RNA 双链 DNA 模板,构建表达质粒 pGenesil-2/CCND2,转染 LP-1细胞,以 RT-PCR 检测对 CCND2表达的抑制作用,以锥虫蓝染色计数和 MTF 检测细胞增殖,以 Annexin V 检测细胞凋亡,流式细胞术榆测细胞周期。结果质粒 pGenesil-2/CCND2转染 LP-1细胞.转染率为34.2%,可以明显抑制 CCND2的表达,细胞增殖明显受抑制,72h后细胞凋亡率为(25.7+4.8)%。结论抑制 CCND2的表达可以抑制骨髓瘤细胞生长,并诱发骨髓瘤细胞的凋亡,可能是潜在的治疗靶点。

The effect of cyclin D2 shRNA on the proliferation and apoptosis of LP-1 cell

OBJECTIVE: To construct cyclin D2 (CCND2) short hairpin RNA ( shRNA) plasmid for repressing the expression of CCND2 in human myeloma cell line LP-1,and to detect its effect on the proliferation and apoptosis of LP-1 cell. METHODS: A CCND2 shRNA model was constructed and cloned into plasmid pGensil-2, then the plasmid was transfected into LP-1 cell in vitro. The CCND2 expression cell proliferation, cell cycle and cell apoptosis of the transfected LP-1 cells were studied by RT-PCR, trypanosome staining, flow cytometry and annexin V assay. RESULTS: The transfection efficiency of LP-1 cell was 34. 2%. In the transfected LP-1 cell CCND2 mRNA expression was reduced significantly, the cell growth was inhibited significantly and the cell cycle was partly arrested in G, phase. The apoptosis rate of the transfected LP-1 cell after 72 h was (25.7+/-4.8)%. CONCLUSION: The inhibition of CCND2 in LP-1 cells could inhibit the cell growth and induce cell apoptosis. CCND2 maybe a new therapeutic target.