沉默B3GAT3 mRNA表达对多发性骨髓瘤细胞增殖的影响

Effect of silencing B3GAT3 mRNA expression on proliferation of multiple myeloma and its mechanism

目的分析B3GAT3 mRNA在MM细胞中的表达变化,探讨沉默β-1,3-葡萄糖醛酸基转移酶3(B3GAT3)mRNA表达对人多发性骨髓瘤(MM)细胞增殖的影响。方法应用GEO数据库的GSE47552数据集,分析41例MM患者和5例正常人骨髓浆细胞B3GAT3 mRNA表达差异。选择UCSC XENA数据库中的858例MM患者,根据MM细胞B3GAT3 mRNA表达量的中位值将患者分为高表达和低表达,比较不同B3GAT3 mRNA表达水平患者的预后差异。利用CCLE数据库筛选高表达B3GAT3 mRNA的骨髓瘤细胞,选取B3GAT3 mRNA表达水平较高的RPMI8226细胞进行后续实验。将RPMI8226细胞分为空白对照组、sh-NC组和sh-B3GAT3组,sh-B3GAT3组和sh-NC组分别转染B3GAT3-shRNA慢病毒、对照慢病毒,采用荧光定量PCR法检测细胞B3GAT3 mRNA,采用CCK-8法检测三组细胞增殖情况。结果GEO数据库结果显示,MM患者骨髓浆细胞B3GAT3 mRNA表达水平高于正常人(P<0.05)。UCSC XENA数据库结果显示,B3GAT3 mRNA高表达患者总体生存时间较低表达患者缩短(P<0.05)。细胞实验显示,sh-B3GAT3组B3GAT3 mRNA表达水平较空白对照组和sh-NC组降低(P均<0.05);培养48、72 h后,sh-B3GAT3组细胞增殖OD值较空白对照组和sh-NC组降低(P均<0.05)。结论B3GAT3 mRNA在MM细胞中表达升高,且该基因高表达的MM患者预后不良;沉默B3GAT3 mRNA表达可抑制MM细胞增殖。

Objective To analyze the expression changes ofβ-1,3-glucuronidyltransferase 3(B3GAT3)mRNA in multiple myeloma(MM)cells,and to investigate the effect of silencing B3GAT3 mRNA expression on proliferation of human MM cells.Methods The GSE47552 dataset from the GEO database was used to analyze the difference in B3GAT3 mRNA expression between the bone marrow MM cells from 41 patients and normal plasma cells from 5 healthy individuals.Totally 858 MM patients in the UCSC XENA database were collected,and the patients were divided into the high expression group and low expression group based on the median value of MM cells B3GAT3 mRNA expression,and the prognostic differences between patients in these two groups were compared.The CCLE database was used to screen the MM cells with high expression of B3GAT3 mRNA,and RPMI8226 cells were selected for subsequent experiments.RPMI8226 cells were divided into the blank control group,sh-NC group,and sh-B3GAT3 group,respectively.Cells in the sh-B3GAT3 group and the sh-NC group were transfected with the B3GAT3-shRNA lentivirus and control lentivirus.B3GAT3 mRNA was detected by fluorescence quantitative PCR.The cell proliferation was detected by the CCK-8 method.Results The GEO database results showed that the expression level of B3GAT3 mRNA in MM group was significantly higher than that in normal plasma cell group(P<0.05).The UCSC XENA database results found that patients with high B3GAT3 expression had shorter overall survival time compared with the low expression group(P<0.05).Cell experiments exhibited that the sh-B3GAT3 group had lower B3GAT3 mRNA expression level than the blank control group and the sh-NC group(both P<0.05).At 48 and 72 h of culture,the cell proliferation OD value decreased in the sh-B3GAT3 group in comparison with those of the blank control group and the sh-NC group(both P<0.05).Conclusions The expression of B3GAT3 is upregulated in MM cells,and the prognosis of MM patients with high expression of this gene is poor.Silencing B3GAT3 expression can inhibit the proliferation of MM cells.