摘要
目的 观察RhoC蛋白在骨髓瘤组织中的表达及其对多发性骨髓瘤细胞株RPMI8226细胞增殖、凋亡的影响.方法 利用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶（SP）法检测55例多发性骨髓瘤组织及7例骨髓组织中RhoC基因的表达,采用RhoC小干扰RNA （siRNA）转染RPMI 8226细胞,采用逆转录-聚合酶链反应（RT-PCR）、Western blot检测RhoC mRNA和蛋白的表达;分别在转染后24、48、72 h通过噻唑蓝（MTT）法观察RhoC siRNA对细胞增殖的影响;转染后加入膜联蛋白V（Annexin V）和碘化丙锭（Pl）检测RhoC siRNA对细胞凋亡的影响.结果 与对照组比较,RhoC基因在多发性骨髓瘤组织中高表达（P＜0.05）;RhoC siRNA转染细胞72 h后,RhoC mRNA和蛋白的表达量分别为1.90±1.03、0.22±0.03,与其对照组比较均明显降低（P＜0.05）;细胞生长抑制率[（89.02±7.03）％]及早期凋亡细胞比例[（12.02±1.12）％]明显高于对照组（P＜0.05）.结论 RhoC siRNA具有抑制RPMI 8226细胞增殖,促进凋亡的作用.
Objective To investigate the expression of RhoC in multiple myeloma tissues and influence of RhoC gene on proliferation and apoptosis of human multiple myeloma RPMI ＋ 8226 cells.Methods Immunohistochemical streptavid-in-peroxidase （SP） methods were used to detect the expression levels of RhoC in 55 cases of multiple myeloma tissues and 7 cases of myeloid tissues.The small interfering RNA （siRNA） was transfected into human multiple myeloma RPMI ＋ 8226 cells.Reverse transcriptasepolymerase chain reaction （RT-PCR） and Western blotting were used to detect the levels of RhoC mRNA and protein.The effect of RhoC siRNA on cell proliferation was measured by methyl thiazol tetrazolium （MTT） assay at 24,48 and 72 h after transfection.After transfection Annexin V/propidium iodide （PI） double staining method was used to detect effects of RhoC siRNA on cell apoptosis.Results （1） As compared with control group,the RhoC gene was overexpressed in multiple myeloma group （P 〈 0.05）.（2） As compared with control groups,after transfection of RhoC siRNA for 72 h,the levels of RhoC mRNA and protein were 1.90 ± 1.03 and 0.22 ± 0.03,which were knocked down significantly （P 〈 0.05）,and the inhibition rate of cell proliferation （89.02 ±7.03） % and the number of apoptosis cells （12.02 ± 1.12） % were greatly increased （P 〈 0.05）.Conclusion RhoC siRNA can inhibit proliferation and promote cell apoptosis of human multiple myeloma RPMI ＋ 8226 cells.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第12期2799-2801,共3页
Chinese Journal of Experimental Surgery
基金
郑州大学第一附属医院青年创新基金资助项目
