趋化因子CCL20在埃罗妥珠单抗及来那度胺耐药多发性骨髓瘤细胞中差异表达的研究

Research of differential expression of chemokine CCL20 in elotuzumab and lenalidomine-resistant multiple myeloma cells

目的:研究趋化因子CCL20是否与多发性骨髓瘤细胞对埃罗妥珠单抗及来那度胺耐药相关。方法:构建埃罗妥珠耐药及埃罗妥珠与来那度胺双耐药的细胞模型;Western blot检测来那度胺敏感(U266/WT,UW)、来那度胺耐药(U266/R10R,UR)、埃罗妥珠耐药(U266/Elo,UWE)及埃罗妥珠与来那度胺双耐药(U266/R10R/Elo,URE)的U266细胞系中CS1蛋白的表达;基因表达谱芯片检测四种细胞mRNA差异表达;RT-PCR验证四种细胞CCL20基因的mRNA差异表达。结果:成功构建了埃罗妥珠耐药与埃罗妥珠及来那度胺双耐药骨髓瘤细胞系;四种细胞中埃罗妥珠单抗作用的靶点CS1蛋白表达无明显差异;基因表达芯片显示在埃罗妥珠及来那度胺双耐药细胞中,CCL20基因的表达显著下调;RT-PCR证实,与来那度胺敏感株相比,CCL20 mRNA在三种耐药株中均有不同程度下降,其中在双耐药细胞株中表达下降最明显。结论:CCL20在来那度胺与埃罗妥珠单抗耐药骨髓瘤细胞株中表达下降,可能与耐药有关。

Objective:To investigate the different expression of chemokine CCL20 in the multiple myeloma cells which were resistant to elotuzumab and lenalidomide.Methods:The cell models of elotuzumab resistance and elotuzumab and lenalidomide resistance were established.Western blot was used to detect the expression of CS1 protein in U266 cell line of lenalidomide sensitive(U266/WT,UW),lenalidomide resistant(U266/R10R,UR),elotuzumab resistant(U266/Elo,UWE),elotuzumab and lenalidomide resistant(U266/R10R/Elo,URE).The mRNA expression of CCL20 gene in four kinds of cells was detected by RT-PCR.The difference of mRNA expression in four kinds of cells was detected by gene expression microarray.Results:The myeloma cells resistant to elotuzumab,elotuzumab and lenalidomide were successfully constructed,and there was no significant difference in the expression of CS1 protein among the four kinds of cells.Gene expression micro array showed that the expression of CCL20 gene was significantly downregulated in elotuzumab and lenalidomide resistant cells.RT-PCR confirmed that compared with lenalidomide-sensitive strains,the mRNA of CCL20 decreased in different degrees in the three drug-resistant cell lines,especially in the elotuzumab and lenalidomide resistant cell lines.Conclusion:The decreased expression of CCL20 in lenalidomide and elotuzumab resistant myeloma cell lines may be related to drug resistance.