微RNA-575在胰腺癌干细胞中的表达  

Expression of miR-575 in pancreatic cancer stem cells

作　　者：宫伟强[1] 杜福田[1] 丁维宝[1] 林洪峰[1] 董承伟[1] 

机构地区：[1]潍坊市人民医院肝胆外科,潍坊261041

出　　处：《中华肝胆外科杂志》2013年第12期930-933,共4页Chinese Journal of Hepatobiliary Surgery

摘　　要：目的应用无血清培养基培养胰腺癌干细胞，检测其微RNA-575（miR-575）的表达。方法应用无血清培养基培养ASPC—l和PANC-1细胞系，分离成球样悬浮生长的细胞球群作为胰腺癌干细胞，并通过检测侵袭、成瘤能力和表面标记物CD24／CD44表达等验证其，肿瘤干细胞特性。荧光定量逆转录PCR技术检测miR-575在胰腺癌干细胞中的表达。结果ASPC-1和PANC-1细胞系中分别有一小部分细胞能在无血清培养基中存活增殖，形成悬浮的肿瘤细胞球。ASPC-1和PANC-1细胞球的平均穿膜细胞数分别为（147．3±18．6）个和（113．2±12．9）个，较细胞系具有较强的侵袭能力。ASPC-1和PANC1细胞球移植形成肿瘤所需最少细胞数为5×10-5个，其成瘤能力是普通细胞系的100倍。ASPC-1、PANC1细胞球CD24＋CD44＋比例分别为0．38％～0．43％、4．91％～5．21％，高于细胞系中的表达（P〈O．05）。ASPC-1和PANC-1细胞球miR575表达量分别是胰腺癌细胞系的3．71、3．83倍，表达上调（P〈O．05）。结论应用无血清培养基可以从ASPC-1和PANC-1细胞系中分离出少量具有干细胞特性的胰腺癌细胞球。其miR-575表达上调，可能是胰腺癌干细胞特性维持关键基因。Objective To detect the expression level of miRNA-575 by pancreatic cancer stem cells. Methods Pancreatic cancer cell lines were cultured with serum free medium. The cancer stem cells were obtained and their characteristics were observed with an invasion assay, a xenograft experi merit, and flow cytornetry analysis. Moreover, qRT-PCR was used to detect the expression of miR- 575. Results A small subpopulation of cells in both ASPC 1 and PANC-1 cells survived and formed spheres in the absence of serum. The mean number of migrated cells of PANC 1 and ASPC1 spheres was （147.3±18.6） cells and （113.2±12.9） cells. Therefore, the spheres have higher invasive ability compared with those of parental cells. As few as 5 × 105 cells in PANC-1 and ASPC-1 spheres could initiate a xenograft tumor in NOD/SCID mice, which suggested the tumorigenicity of the spheres was 100 times those of parental cell lines. The percentages of CD24＋CD44＋ in spheres were 0.38% -0.43o/00 and 4.91%-5.21%, which showed increased expression compared with ASPC-1 and PANC- 1 ceil lines （P〈0.05）. The expression of miR 575 in spheres was 3.71 and 3.83 times its expression in cell lines ASPC 1 and PANC1 respectively （P〈0.05）. Conclusions Pancreatic cancer cell spheres may be a source for collecting cancer stem cells in a serum-free medium culture, and MiR 575 may play an important role in regulating the biological characteristics of pancreatic cancer stem cells.

关 键 词：胰腺肿瘤 肿瘤干细胞 微RNAS 

分 类 号：R735.9[医药卫生—肿瘤]