高转移性人乳腺癌细胞亚克隆的筛选  

Screening of subclones of high metastatic human breast cancer cells

作　　者：潘秀华[1,2] 王立梅[1] 李希[1] 冯玉梅[1] 

机构地区：[1]天津医科大学附属肿瘤医院生物化学及分子生物学室教育部乳腺癌防治重点实验室,天津300060 [2]徐州医学院机能科研实验中心,江苏徐州221004

出　　处：《中华乳腺病杂志（电子版）》2010年第1期53-56,共4页Chinese Journal of Breast Disease(Electronic Edition)

基　　金：国家自然科学基金项目(30471671;30872518);天津市自然科学基金重点项目(09JCZDJC19800);教育部长江学者和创新团队发展计划项目(IRT0743)

摘　　要：目的筛选高转移潜能的人乳腺癌细胞亚克隆,为人类乳腺癌转移相关的体内外研究提供实验对象。方法通过有限稀释法分离和培养人乳腺癌细胞MDA-MB-435S的单克隆;采用细胞生物学方法体外评价亚克隆的克隆形成、增殖、运动和侵袭能力;将MDA-MB-435S亚克隆细胞通过乳腺脂肪垫接种于免疫双缺陷SCID鼠,验证体外筛选克隆的体内转移能力。定量资料采用独立样本t检验进行统计学分析。结果筛选得到MDA-MB-435S的高转移亚克隆14-E5;14-E5与亲代细胞呈梭形、伪足较长的细胞形态明显不同,体积比亲代细胞小,呈多边形,触角增多;细胞的体外克隆形成能力、运动能力、侵袭能力及体内自发转移能力均较亲代显著增强(P<0.050);细胞周期S期和G2/M期比例比亲代减少,增殖能力较亲代降低(t=7.047,P=0.002);细胞异质黏附能力、体内成瘤率与亲代之间的差异无统计学意义(P>0.050)。结论筛选并建立了高转移潜能人乳腺癌细胞亚克隆细胞株14-E5,可以用于乳腺癌转移基因筛选、转移机制研究、抗转移药物的研发和评价抗转移实验性治疗疗效。Objective To isolate subclones of high metastatic potential from human breast cancer cell line MDA-MB-435S in the interest of researches on metastasis of breast cancer in vitro and in vivo. Methods Cell monoclones with different metastatic potential were isolated from their parental cell line MDA-MB-435S by limiting dilution. Then the cloning formation, proliferation, invasion and migration abilities of these subelones were identified in vitro via biology methods. Subclone with high metastasis capability was verified by injecting MDA-MB-4355 subclone cells into fat pad of SCID mice to observe the spontaneous metastasis ability in vivo. Quantitative data were analyzed using independent samples t test. Results A subelone with high metastatic capability 14-E5 from MDA-MB-435S was isolated. 14-E5 subclone was polygon, with more pseudopodiums and smaller than MDA-MB-435S which is fusiform shape with long pseudopodium. The clonality of 14-E5 was stronger than parental cell line MDA-MB- 435S （t=3.720,P=0.003）. The motility （t=5.575,P=0.005）, invasion capability （t=9.177,P= 0.00l）, and the spontaneous metastasis capacity of 14-E5 were also stronger than MDA-MB-435S. The proportion of cells in DNA synthesis and mitosis states was lower in 14-E5 than in MDA-MB-435S （ t= 7. 047,P=0. 002）. No significant differences of heterogeneity adhesion capacity and tumorigenicity were found between 14-E5 and MDA-MB-435S （P〉0. 050）. Conclusions 14-E5 subclone with high metastatic capability will be valuable in screening breast cancer metastatic gene, investigating the mechanisms of metastasis, exploring anti-metastasis medicine, and supervising experimental therapeutic efficacy of antimetastasis.

关 键 词：乳腺肿瘤 转移 亚克隆 

分 类 号：R737.9[医药卫生—肿瘤]