人卵巢癌OVCAR3细胞系中侧群细胞的分离及其成瘤性、侵袭性的实验研究  

Tumorigenicity and invasion of side population in human ovarian epithelial cancer OVCAR3 cells

作　　者：王伟[1] 王沂峰[1] 张岭梅[1] 林琼燕[1] 黄菊[1] 

机构地区：[1]广州医学院第三附属医院妇产科,广州510150

出　　处：《山东大学学报（医学版）》2009年第6期8-11,共4页Journal of Shandong University:Health Sciences

基　　金：广东省自然科学基金资助课题(7002691);广东省社会发展领域科技计划项目(53);广东省名医工程(30)

摘　　要：目的分离卵巢癌细胞系OVCAR3中的侧群(SP)细胞,并观察其成瘤性和侵袭性。方法利用流式细胞荧光激活分选法将卵巢癌OVCAR3细胞系分成SP和非侧群(non-SP)细胞两个亚群。对两个亚群细胞分别采用软琼脂克隆形成实验和细胞球形成实验观察其体外成瘤能力;Transwell小室法检测两个亚群细胞的体外侵袭力。结果OVCAR3细胞株中外排Hoechst33342的SP细胞占13.1%,加入维拉帕米SP细胞的比例为1.8%。SP细胞的体外克隆形成率为(46.17±23.27)%,non-SP细胞仅为(6.17±3.06)%,SP细胞克隆形成率高于non-SP细胞(P<0.05);细胞球形成实验结果显示,SP细胞能在无血清的培养液中形成细胞球,而non-SP细胞则不能;Transwell侵袭实验结果显示,SP表型细胞穿过Matrigel基底膜的细胞数为(215.9±30.9)个,non-SP表型细胞仅为(60.9±10.5)个。结论卵巢癌OVCAR3细胞系中存在外排Hoechst33342荧光染料的SP细胞,其成瘤性、侵袭力均强于non-SP亚群细胞,表明SP表型的肿瘤细胞在卵巢癌的生长、侵袭及转移中具有重要的地位。Objective To isolate side population（SP） cells from human ovarian epithelial cancer OVCAR3 cells and to study their tumorigenicity and invasion. Methods Flow cytometry and Hoechst 33342 dye efflux assay were used to isolate SP cells and non-SP cells from the OVCAR3 cell line of human ovarian epithelial carcinoma. Tumorigenicity of the two subpopulations was observed by a soft agar cloning method and sphere-forming assay. Invasiveness was measured by a Transwell experiment. Resulls The proportion of SP cells excluding Hoechst 33342 dye in the OVCAR3 cells was 13.1%, by addition of verapamil, it was 1.8%, which was greatly decreased. CFE of SP and non-SP was 46.17% ± 23.27% and 6.17% ± 3.06% ,respeetively（P 〈 0.05）. SP cells formed cell spheres in serum-free culture, but non-SP cells did not. The cell number that infiltrated the Transwell membrane in the SP group（215.9 ± 30.9） was significantly higher than that in the non-SP group （60.9 ± 10.5）. Conclusion Human epithelial ovarian carcinoma cell line OVCAR3 contains a small subpopulation of cells excluding Hoeehst 33342 dye that exhibits high tumorigenicity and invasion potential, indicating that SP cells have played a principal role in ovarian carcinoma cell proliferation and invasiveness.

关 键 词：卵巢肿瘤 肿瘤干细胞 侧群细胞 肿瘤侵润 

分 类 号：R737.31[医药卫生—肿瘤]