目的 观察Notch1信号通路活化与多发性骨髓瘤(MM)发生、发展及耐药的关系,为MM治疗提供新靶点.方法 利用携带Notch1-1CN逆转录病毒载体及Notch1-对照病毒(CON)载体感染RPMI8226细胞(分别为病毒组及病毒对照组),同时设未经感染的细胞为未转染组,采用Western blot法检测Notch1-ICN蛋白表达鉴定细胞；采用CCK-8法检测细胞增殖情况及对硼替佐米敏感性的影响；流式细胞术检测细胞周期.结果 病毒组Notch1 -ICN表达明显上调,约为病毒对照组和未转染组的2.5倍；培养后48、72、96h病毒组细胞增殖率均显著高于病毒对照组(D均＜0.01)；随硼替佐米浓度增高,各组细胞抑制作用均逐渐增加,且同浓度条件下病毒组抑制率显著低于病毒对照组及未转染组(P＜0.05或0.01)；病毒组S期细胞比例明显高于其余两组(P均＜0.01).结论Notch1信号通路活化与MM的发生、发展及耐药密切相关,有望成为MM治疗的新靶点.%Objective To investigate the relationship between Notchi activation and the occurrence, development and drug resistance of multiple myeloma(MM), so to provide the basis for molecular targeted therapy. Methods Human RP-MI8226 MM cells were infected with Notchl-ICN and Notchl-CON retrovirus stocks ( Notchl-ICN group and Notchl-CON group). RPMI8226 cells without infection were defined as unlransfected group. Notchl-ICN protein expression was detected by Western blot analysis. CCK-8 was used to detect the proliferation of MM cells and sensitivity to chemotherapeutic drugs. Cell cycle analysis was detected by flow cytometry. Results The relative expression level of Notchl-ICN protein in Notchl-ICN group increased significantly, and was 2.5 fold higher than that in Notchl-CON group or untransfected group, indicating Notchi signal activation in Notchl-ICN group; the cell proliferation rates in Notchl-ICN group at 48 h, 72 h and 96 h were all significantly increased as compared with those in Notchl-CON group or untransfected group (P <0.01) , and the inhibitory rate was decreased in RPMI8226-ICN cells at 48 h treated by the same concentration of Bortezomib ( P < 0.05 or 0.01) ; S phase cells in Nolchl-ICN group were increased as compared with that in Notchl-CON group or untransfected group (P<0.05). Conclusion Notchi activation was closely correlated with the occurrence, development and drug resistance of MM; Notchi signaling is promising to be used as a new target for MM treatment.
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