CAJ | 학술논문

目的：探讨Notch1 siRNA对体内外人骨髓瘤细胞RPMI-8226细胞硼替佐米药物敏感性的影响。方法体外采用Notch1 siRNA转染RPMI-8226细胞，CCK-8实验检测细胞增殖及硼替佐米药物的敏感性；Western blot检测各组细胞Notch1蛋白表达变化；将RPMI-8226细胞皮下注射于NOD/SCID小鼠，建立人多发性骨髓瘤（MM）小鼠移植瘤模型，将成瘤小鼠分为三组：NS＋bortezomib（Notch1 siRNA转染联合硼替佐米）组；CS＋bortezomib（Control siRNA转染联合硼替佐米）组；UN＋bortezomib（硼替佐米）组，观察各组肿瘤体积变化，免疫组化染色法观察Notch1变化，TUNEL法检测细胞凋亡。结果 Notch1 siRNA有效下调骨髓瘤细胞RPMI-8226细胞Notch1蛋白表达；Notch1 siRNA在96 h抑制细胞增殖作用明显增加，与CS及UN组比较对细胞增殖的作用可见明显差异（P＜0.01）；Notch1 siRNA 转染组细胞对硼替佐米 IC50值为1.21μmol/L，与Control siRNA 转染组及未转染组相比均存在统计学差异（P＜0.01）；Notch1 siRNA 降低移植瘤Notch1蛋白表达，Notch1 siRNA转染组细胞的AI明显高于Control siRNA转染组及未转染组（P＜0.01）；Notch1 siRNA转染联合硼替佐米组肿瘤体积明显减小，13、17及21 d与Control siRNA转染联合硼替佐米组比较均有统计学差异（P＜0.05）。结论体外实验Notch siRNA抑制人骨髓瘤细胞RPMI-8226细胞增殖增加硼替佐米的敏感性，体内试验证实Notch siRNA联合硼替佐米可以明显减小荷瘤MM小鼠肿瘤体积、增加凋亡，提示Notchl是治疗MM的有效分子靶点。
목적：탐토Notch1 siRNA대체내외인골수류세포RPMI-8226세포붕체좌미약물민감성적영향。방법체외채용Notch1 siRNA전염RPMI-8226세포，CCK-8실험검측세포증식급붕체좌미약물적민감성；Western blot검측각조세포Notch1단백표체변화；장RPMI-8226세포피하주사우NOD/SCID소서，건립인다발성골수류（MM）소서이식류모형，장성류소서분위삼조：NS＋bortezomib（Notch1 siRNA전염연합붕체좌미）조；CS＋bortezomib（Control siRNA전염연합붕체좌미）조；UN＋bortezomib（붕체좌미）조，관찰각조종류체적변화，면역조화염색법관찰Notch1변화，TUNEL법검측세포조망。결과 Notch1 siRNA유효하조골수류세포RPMI-8226세포Notch1단백표체；Notch1 siRNA재96 h억제세포증식작용명현증가，여CS급UN조비교대세포증식적작용가견명현차이（P＜0.01）；Notch1 siRNA 전염조세포대붕체좌미 IC50치위1.21μmol/L，여Control siRNA 전염조급미전염조상비균존재통계학차이（P＜0.01）；Notch1 siRNA 강저이식류Notch1단백표체，Notch1 siRNA전염조세포적AI명현고우Control siRNA전염조급미전염조（P＜0.01）；Notch1 siRNA전염연합붕체좌미조종류체적명현감소，13、17급21 d여Control siRNA전염연합붕체좌미조비교균유통계학차이（P＜0.05）。결론체외실험Notch siRNA억제인골수류세포RPMI-8226세포증식증가붕체좌미적민감성，체내시험증실Notch siRNA연합붕체좌미가이명현감소하류MM소서종류체적、증가조망，제시Notchl시치료MM적유효분자파점。
Objective To study the effect of Notch1 siRNA on human myeloma cell line RPMI-8226 cells to bortezomib sensitivity in vitro and in vivo. Methods Notch1 siRNA was transfected into RPMI-8226 cells, and cell proliferation and drug sensitivity was detected by CCK-8 assay. The expression of Notch1 protein was detected by Western blot. RPMI-8226 cells were subcutaneously implanted in NOD/SCID mice, the mice were divided into three groups:NS+bortezomib (Notch1 siRNA transfection combined with bortezomib) group; CS+bortezomib (Control siRNA transfection in combination with bortezomib) group;UN+bortezomib (bortezomib) group. The changes of tumor volume were observed. The immunohistochemical staining method was used to observe the expressions of Notch1. TUNEL was used to detect apoptosis. Results Notch1 siRNA can effectively inhibit the expression of Notch1 protein in myeloma RPMI-8226 cells;Notch1 siRNA significantly inhibited cell proliferation at 96 hours, and there were obvious difference compared with the CS and UN groups on the cell proliferation (P<0.01); IC50 value of bortezomib in NS group was 1.21 μmol/L, and there were obvious difference compared with the CS and UN groups (P<0.01);Notch1 siRNA decreased expression of Notch1 protein in tumor cells; AI in NS group was significantly higher than that of CS group and non transfection group (P<0.01); Tumor volume in Notch1 siRNA transfection in combination with bortezomib group was significantly reduced at 17 d, 13 d, and 21 d, compared with control siRNA transfection in combination with bortezomib group (P<0.05). Conclusion In vitro, Notch siRNA inhibits the proliferation of human myeloma RPMI-8226 cells, and increases bortezomib sensitivity. In vivo, tumor volume in Notch1 siRNA transfection in combination with bortezomib group is significantly reduced, and the apoptosis in NS group is increased, suggesting that Notchl is an effective molecular target in the treatment of myeloma.