CAJ | 학술논문

目的研究癌基因B-RafV600E导致肿瘤细胞染色体不稳定的分子机制.方法采用RNA干扰技术敲除稳定表达B-RafV600E基因的黑色素瘤Sbcl2和SK-MEL31细胞中内源性单核纺锤体蛋白激酶(Mps1)基因表达,免疫荧光染色技术检测中心体及纺锤体结构.HU-arrest分析法观察Mps1基因缺失对癌基因B-RafV600E致肿瘤细胞中心体过度复制及多极纺锤体形成的影响.结果未敲除内源性Mps1基因的表达B-RafV600E基因的Sbcl2和SK-MEL31细胞中36％出现中心体过度复制及多极纺锤体,Mps1基因被敲除后上述异常细胞比例降低至6％.结论 B-RafV600E可能通过Mps1调控中心体过度复制及多极纺锤体结构的形成,影响肿瘤细胞染色体不稳定性及非整倍体细胞的出现.
목적 연구암기인B-RafV600E도치종류세포염색체불은정적분자궤제.방법 채용RNA간우기술고제은정표체B-RafV600E기인적흑색소류Sbcl2화SK-MEL31세포중내원성단핵방추체단백격매(Mps1)기인표체,면역형광염색기술검측중심체급방추체결구.HU-arrest분석법관찰Mps1기인결실대암기인B-RafV600E치종류세포중심체과도복제급다겁방추체형성적영향.결과 미고제내원성Mps1기인적표체B-RafV600E기인적Sbcl2화SK-MEL31세포중36％출현중심체과도복제급다겁방추체,Mps1기인피고제후상술이상세포비례강저지6％.결론 B-RafV600E가능통과Mps1조공중심체과도복제급다겁방추체결구적형성,영향종류세포염색체불은정성급비정배체세포적출현.
Objective To explore the molecular mechanism of BRAFV600E inducing chromosome instability in Sbcl2 and SK-MEL31 melanoma cells.Methods The endogenous Mps1 in stable Sbcl2-and SK-MEL31-B-RafV600E expression cells were depleted by siRNA approach.To test the effect of B-RafV600E on the centrosome amplification and the formation of multipolar spindles,cells at S-phase with HU-treatment were arrested and then the centrosomes and mitotic spindles structure were detected through immunofluoresence.Results The percentage of B-RafV600E expressing Sbcl2 and SK-MEL31 cells (Sbcl2-B-RafV600E and SKMEL31-B-RafV600E) with centrosome amplification and multipolar spindle was reduced from 36 ％ to 6 ％ when Mps1 was absent.Conclusion B-RafV600E leads to centrosome amplification and multipolar spindle through Mps1,thus results in chromosome instability in Sbcl2 and SK-MEL31 melanoma cells.