干细胞标志物在卵巢癌SKOV3细胞侧群细胞中的表达  

Expression of stem cell markers in the side population of ovarian cancer SKOV3 cells

作　　者：钟艳平[1] 孟泳圳[2] 苏节[1] 李力[1,2] 黎丹戎[1] 张玮[2] 

机构地区：[1]广西医科大学医学科学实验中心,广西南宁530021 [2]广西医科大学附属肿瘤医院妇瘤科,广西南宁530021

出　　处：《中国肿瘤生物治疗杂志》2014年第5期521-525,共5页Chinese Journal of Cancer Biotherapy

基　　金：广西卫生厅重点项目(No.2010074);广西自然科学基金资助项目(No.2012GXNSFAA053134);广西研究生教育创新计划项目资助(No.YCSZ2012045)~~

摘　　要：目的:检测干细胞相关标志物在卵巢癌SKOV3细胞系侧群(side population,SP)细胞和非侧群(Non-SP)细胞中的表达差异,确定卵巢癌干细胞特异性标志物。方法:Hoechst 33342染色检测SKOV3细胞中SP细胞的比例,流式细胞术检测SKOV3细胞的SP和Non-SP细胞中干细胞标志物CD133、CD117、CD44、ABCG2、ALDH1的表达情况,荧光定量PCR检测SP和Non-SP细胞中ABCG2、ALDH2、NANOG、OCT4、SOX2、CD133、CD117 mRNA的表达水平。结果:SKOV3细胞中SP细胞比例为(1.56±0.35)%。ALDH1、ABCG2在SP细胞中表达率分别为(87.3±5.76)%、(29.48±4.43)%,在Non-SP的表达率分别为(5.32±0.47)%、(3.01±1.69)%,差异有统计学意义(P<0.01);CD44在这两种细胞亚群中的表达率均高于99%(P>0.05);CD133、CD117在这两种细胞亚群中均不表达。ALDH1、ABCG2、NANOG、OCT4和SOX2 mRNA在SP细胞中的表达分别是Non-SP细胞的21.03倍(P=0.001)、3.14倍(P=0.001)、23.94倍(P=0.001)、10.73倍(P=0.009)和21.46倍(P=0.001),CD133、CD117 mRNA在两种细胞亚群中均不表达。结论:人卵巢癌细胞株SKOV3中存在SP细胞亚群,ALDH1、ABCG2和NANOG、OCT4、SOX2 mRNA可能是卵巢癌干细胞标志物,为诊断及治疗卵巢癌提供了潜在的靶点。Objective： To study the differential expression of ovarian cancer stem cell specific markers in the side population（ SP） and non-SP of human epithelial ovarian cancer SKOV3 cells. Methods： The SP of SKOV3 cells was identified and sorted by flow cytometry using the blue fluorescent dye Hoechst 33342. The expression of CD133,CD117,CD44,ABCG2 and ALDH2 at the protein level in SP and non-SP cells was assessed by flow cytometry. The expression of ALDH1,ABCG2,NANOG,OCT4,SOX2,CD133 and CD117 at the mRNA level in SP and non-SP cells was assessed by RT-PCR. Results： In SKOV3 cells,SP accounted for（ 1. 56 ± 0. 35） %. ALDH1 and ABCG2 proteins were detected in（ 87.3 ±5.76） % and（ 29. 48 ±4. 43） % of SP cells respectively,both significantly higher than those in non-SP cells（ P 〈0.05）. CD44 protein was detected in greater than 99% of cells in both subpopulations（ P〉 0.05）. Neither CD133 nor CD117 protein was detected in both subpopulations. The mRNA levels of ALDH1,ABCG2,NANOG,OCT4 and SOX2 in SP cells were increased by 21. 03（ P = 0. 001）,3. 14（ P = 0. 001）,23. 94（ P = 0. 001）,10. 73（ P = 0. 009）,21. 46（ P =0.001） folds,respectively,as compared with the levels in Non-SP cells. CD133 and CD117 transcripts were detected in neither sub-population of SKOV 3 cells. Conclusion： There is a SP in human ovarian cancer SKOV3 cells expressing stem cell markers ALDH1 and ABCG2 at the protein level and NANOG,OCT4,and SOX2 at the mRNA level like ovar-ian cancer stem cells. This SP of SKOV3 cells may offer a very useful tool for ovarian cancer stem cell research.

关 键 词：卵巢癌 SP细胞 肿瘤干细胞 标志物 

分 类 号：R737.31[医药卫生—肿瘤] R730.2[医药卫生—临床医学]