人宫颈癌侧群细胞差异微小RNA表达分析

目的筛选入宫领癌侧群细胞差异表达的微小RNA（microRNA），方法选取16例行宫颈鳞癌切除的手术标本，进行宫颈癌细胞原代培养，流式细胞仪分选侧群细胞，对比侧群细胞和非侧群细胞的体外自我更新能力和肿瘤形成能力。分别提取侧群细胞和非侧群细胞的总RNA，用microRNA芯片分刖检测侧群细胞和非侧群细胞的micl’oRNA表达谱并进行分析，筛选出差异表达的microRNAi．结果在原代培养的16株宫颈癌细胞中均分选出侧群细胞，比例占（1．67±0．94）％。培养14d后，侧群细胞的克隆形成牢为0．51±0．07，而非侧群细胞的克隆形成率仪为0．05±0．02，侧群细胞和非侧群细胞的克隆形成率比较差异有统计学意义（t=2．828，P〈0．01）．仅1×10^4个侧群细胞即可形成肿瘤，而至少1×10^6个非侧群细胞才能形成肿瘤。microRNA芯片筛选出表达差异在10倍以上的有16条microRNA，其中3条microRNA上测，13条microRNA下调；差异表达超过共同2倍的microRNA有7条，其中3条microRNA上调，4条microRNA下调，结论人宫颈癌侧群细胞具有肿瘤干细胞特性，人宫颈癌侧群细胞表达一些特异的microRNA，

Expression of microRNA from human cervical cancer side population cells

Objective To screen the expression of microRNA of human cervical cancer side population （SP） cells. Methods The human cervical cancer cells were obtained from fresh human cervical cancer tissue among 16 patients who were diagnosed of cervical cancer. Flow cytometry and Hoechst33342 dye efflux assay were used to isolate SP cells and NSP（ non side population） cells from primaiy cervical cancer cells. Self-renewal and tumorigenic ability of SP anti NSP cells were compared in vitro and in vivo. MicroRNA was isolated from cervical cancer SP an NSP cells. Hybridization was carried out on Sanger miRBase 17.0 Arrays. Screening of microRNA between SP and NSP cells was undertaken Mieroarray Software. Results The SP cells demonstrated a higher clo- ning ability in vitro and a stronger tumorigenesis ability compared with NSP cells in nude mice. 16 microRNAs expression were tbund in SP cells, 10 times more than thai in NSP cells. Three microRNAs were up-regulated and 13 microRNAs were down-regulated in SP cells. 3 microRNAs were up-regulated and 4 microRNAs were down-regulated in SP cells. Conclusions Human cervical cancer SP cells have the characters of cancer stem cells. Some speeifie miemRNAs are found between human cervical cancer SP cells and NSP cells.