Ad-NK4增强人多发性骨髓瘤RPMI8226细胞对硼替佐米化疗敏感性的作用

Ad-NK4 Enhances the Chemosensitivity of Human Multiple Myeloma RPMI 8226 Cells to Bortezomib

目的:观察Ad-NK4能否增强RPMI 8226细胞对硼替佐米(BOR)的化疗敏感性。方法:细胞-基质粘附实验和RPMI 8226细胞-ECV 304细胞粘附实验检测Ad-NK4对RPMI 8226细胞粘附的影响;Western blot检测粘附和侵袭相关蛋白MMP2、MMP3、MMP7、VEGF表达的变化;MTT法检测Ad-NK4对细胞增殖的影响;PI流式细胞术检测Ad-NK4对细胞凋亡的影响;Western Blot检测Cleaved caspase-3、BAX和BCL-2蛋白表达水平的变化。结果:两种细胞粘附实验均显示Ad-NK4可显著抑制RPMI 8226细胞粘附,且与Erk信号通路和JAK/STAT信号通路显著相关(P<0.05);Ad-NK4能明显降低RPMI 8226细胞MMP-2、MMP-3和VEGF蛋白的表达水平(P<0.05),但对M M P-7的蛋白表达水平无明显影响(P>0.05)。Ad-NK4和BOR联用的细胞抑制率和诱导凋亡比例均显著高于单用Ad-NK4或BOR。同样,Ad-NK4和BOR联用组Cleaved caspase-3和BAX蛋白水平均明显高于单独作用组,而BCL-2蛋白水平却相反,同时在联用组BAX/BCL-2比率增加。结论:通过活化Caspase-3和上调BAX/BCL-2比率的途径,Ad-NK4可增强BOR体外抗骨髓瘤RPM I 8226细胞作用。

Objective:To investigate if Ad-NK4 can enhance the chemosensitivity of human multiple myeloma RPMI8226 cells to bortezomib(BOR).Methods:The cell-matrix adhesion test and PRMI 8226 cell-EC V 304 cell adhesion test were used to analyze the effect of Ad-NK4 on adhesion of RPMI 8226 cells;Western blot was used to detect the expression changes of adhesion and invasion-associated proteins MMP2,MMP3,MMP7 and VEGF;MTT assay was used to detect the effect of Ad-NK4 on proliferation of RPMI 8226 cells;the flow cytometry with PI staining was used to detect the effect of Ad-NK4 on cell apoptosis;the expression of cleaved caspase-3,BAX and BCL-2 was assayed by Western blot.Results:These 2 adhesion assays indicated that Ad-NK4 significantly inhibited the adhesion of human multiple myeloma RPMI 8226 cells.In addition,Erk and JAK/STAT pathway may be involved in the process.The expression level of MMP-2,MMP-3 and VEGF were decreased in Ad-NK4 group,compared with untreated or Ad-GFP group(P <0.05).However,the expression of MMP-7 protein in Ad-NK4 group was not significantly different from untreated or Ad-GFP group(P >0.05).The inhibitory rates of the proliferation in cells treatedly Ad-NK4 combined with BOR was significantly higher than that with BOR or Ad-NK4 alone.Similarly,Western blot indicated that the level of cleaved caspase-3 and BAX in cells treated with Ad-NK4 combined with BOR was significantly higher than BOR or Ad-NK4 alone,but BCL-2 protein expression was significantly lower.Meanwhile,the ratio of BAX/BCL-2 was increased.Conclusion:Ad-NK4 can enhance the chemosensitivity of human multiple myeloma RPMI 8226 cells to BOR,which is associated with increasing of both BAX/BCL-2 ratio and Caspase-3 activation.