乳腺癌多柔比星耐药细胞的生物学特性  

The biological characteristics of adriamycin-resistant breast cancer cell line

作　　者：孙月[1] 刘瑞娟[2] 庄静[2] 刘丽娟[2] 郝翠[3] 冯福彬[2] 臧传鑫 赵文歌 孙长岗[2] 

机构地区：[1]潍坊医学院临床医学院研究生处,山东潍坊261000 [2]潍坊市中医院肿瘤科,山东潍坊261000 [3]青岛大学医学院脑血管病研究所,山东青岛266000 [4]山东中医药大学中医学院研究生处,山东济南250000

出　　处：《肿瘤》2016年第10期1098-1106,共9页Tumor

基　　金：国家自然科学基金资助项目(编号:81273987;81473513)~~

摘　　要：目的 :探讨乳腺癌多柔比星(adriamycin,ADR)耐药细胞的生物学特性。方法 :以人乳腺癌MCF-7细胞为亲本细胞,建立对ADR耐药的人乳腺癌MCF-7/ADR细胞。应用刃天青法检测ADR对MCF-7/ADR和MCF-7细胞的半数抑制浓度(half maximal inhibitory concentration,IC50)值,应用平板克隆形成实验检测MCF-7/ADR和MCF-7细胞的克隆形成能力。FCM法和免疫荧光法分析MCF-7/ADR和MCF-7细胞中CD44+/CD24-和乙醛脱氢酶1(acetaldehyde dehydrogenase 1,ALDH1)+干细胞亚群所占百分比,蛋白质印迹法检测ALDH1蛋白及调控干细胞的Hedgehog信号通路中Smo(Smoothened)和Gli1蛋白的表达。结果 :ADR对MCF-7/ADR细胞的IC50值明显高于MCF-7细胞(P<0.01),耐药倍数约为385倍。MCF-7/ADR细胞的克隆形成能力显著高于MCF-7细胞(P<0.01)。MCF-7/ADR细胞中CD44+/CD24-和ALDH1+干细胞亚群所占百分比显著高于MCF-7细胞(P值均<0.01)。MCF-7/ADR细胞中ALDH1蛋白及Hedgehog信号通路中Smo和Gli1蛋白的表达水平显著高于MCF-7细胞(P<0.01,P<0.01,P<0.05)。结论:乳腺癌ADR耐药细胞中CD44+/CD24-和ALDH1+干细胞亚群所占百分比以及Hedgehog干细胞调控信号通路相关蛋白Smo和Gli1的表达水平升高,可能与乳腺癌细胞对ADR的耐药相关。Objective： To investigate biological characteristics of adriamycin（ADR）-resistant breast cancer cell line.Methods： Using MCF-7 cells as parent cell line, an ADR-resistant cell line named MCF-7/ADR was established. The half maximal inhibitory concentration（IC50） values of MCF-7/ADR and MCF-7 cells were detected by resazurin test. The ability of colony formation of MCF-7/ADR and MCF-7 cells was measured by colony formation assay. The percentages of CD44^＋/CD24^- and acetaldehyde dehydrogenase 1（ALDH1）＋ subtypes of cancer stem cells in MCF-7/ADR and MCF-7 cells were detected by FCM and immunofluorescent assay, respectively. The expressions of ALDH1 and smoothened（Smo） and Gli1 proteins in Hedgehog signaling pathway related to modulation of stem cells in MCF-7/ADR and MCF-7 cells were detected by Western blotting.Results： The IC50 value of ADR in MCF-7/ADR cells was higher than that in MCF-7 cells（P〈0.01）, the resistant fold was 385. The colony formation rate of MCF-7/ADR was higher than that of MCF-7 cells（P〈0.01）. The percentages of CD44^＋/CD24^- and ALDH1^＋ subtypes of cancer stem cells in MCF-7/ADR cells were higher than those in MCF-7 cells（both P〈0.01）. The expression levels of ALDH1 protein and the Smo and Gli1 proteins in Hedgehog pathway related to modulation of stem cells in MCF-7/ADR cells were higher than those in MCF-7 cells（P〈0.01, P〈0.01 and P〈0.05, respectively）.Conclusion： The percentages of CD44^＋/CD24^- and ALDH1^＋ subtypes of cancer stem cells in MCF-7/ADR cells are higher, and the expressions of ALDH1 protein and the Smo and Gli1 proteins in Hedgehog pathway related to modulation of stem cells in MCF-7/ADR cells are higher. This mechanism may be responsible for ADR resistance in breast cancer cells.

关 键 词：乳腺肿瘤 肿瘤干细胞 抗药性 肿瘤 抗原 CD44 抗原 CD24 ALDH1 HEDGEHOG信号通路 

分 类 号：R737.9[医药卫生—肿瘤]