干扰磷脂爬行酶1基因表达对结直肠癌细胞增殖和粘附的影响  

Influence of silencing PLSCR1 expression with RNA interference on the proliferation and adhesion of colorectal cancer cells

作　　者：崔伟[1] 袁强[1] 陈彩玲[1] 陈纲[1] 李世拥[1] 

机构地区：[1]北京军区总医院普通外科,100700

出　　处：《中华普外科手术学杂志（电子版）》2014年第4期47-49,共3页Chinese Journal of Operative Procedures of General Surgery(Electronic Edition)

基　　金：国家自然科学基金项目(81041025)

摘　　要：目的探讨PLSCR1在结直肠癌细胞增殖和粘附过程中的作用。方法常规培养3株结直肠癌细胞,采用免疫细胞染色和免疫印迹法筛选PLSCR1高表达的细胞株。设计三条RNA干扰片段及一条阴性对照片段,稳定转染高表达PLSCR1的结直肠癌细胞株,采用实时荧光定量PCR和免疫印迹法验证干扰结果,筛选出对PLSCR1蛋白抑制率最高的干扰片段,通过MTT方法来评估转染后结直肠癌细胞增殖能力的变化,采用细胞粘附实验来评估转染后结直肠癌细胞粘附能力的变化。结果免疫细胞染色提示LoVo细胞株高表达PLSCR1,与免疫印迹法结果相一致;转染siRNA-390后,LoVo细胞株PLSCR1的mRNA抑制效果最为明显,抑制率为88.4%,免疫印迹法检测进一步验证siRNA-390片段抑制PLSCR1蛋白表达最为显著。MTT法、纤连蛋白,粘附和层连蛋白检测显示,siRNA-390干扰下调PLSCR1的表达后,细胞生长变缓,粘附能力下降。结论干扰PLSCR1能显著抑制肿瘤的增殖和粘附能力,提示PLSCR1可能在结直肠癌浸润和转移中具有一定作用。Objective To investigate the effect of PLSCR1 on the proliferation and adhesion of colorectal cancer （CRC） cells.Methods Three strains of CRC cells were cultured.The cells with the highest expression of PLSCR1 were harvested for subsequent applications including immunocytochemistry staining and Western-blotting.Three siRNA oligonucleotide segments targeting PLSCR1 were designed.Successful transfection was confirmed by realtime PCR and Western-blotting.The biological behaviors of the cells,including proliferation and adhesion were determined by MTT assay.Results LoVo cells showed the highest expression of PLSCR1 by immunocytochemistry staining and Western-blotting.The siRNA-390 oligonucleotide segment exhibited the best silencing effect in LoVo cells （88.4％） by Western-blotting.After transfection,LoVo cell proliferation was significantly inhibited compared with controls in MTT assay.Ln and Fn adhesion assay showed that the adhesion of LoVo cell was also significantly inhibited.Conclusion By silencing of PLSCR1 using siRNA,the proliferation and adhesion of LoVo CRC cells could be inhibited,suggesting that PLSCR1 might contribute to the tumorigenesis and tumor progression of CRC.

关 键 词：结直肠肿瘤 基因表达 RNA干扰 细胞增殖 细胞粘附 磷脂爬行酶1 

分 类 号：R735.3[医药卫生—肿瘤]