E2通过ER非基因组活性干扰ER乳腺癌细胞对-第三军医大学学报.DOCVIP

非基因组活性对雌激素受体阳性乳腺癌细胞Herceptin敏感性的影响 江晓梅，陈 彬，王 越，王 杨，李渝萍，娄桂予，张 艳，何凤田400038重庆第三军医大学生物化学与分子生物学教研室探讨雌激素（E2）通雌激素受体（ER）非基因组活性干扰ER阳性（ER+ ）乳腺癌细胞对HER2的人源化单克隆抗体Herceptin的敏感性及其机制。方法采用Western blot和瞬时转染结合荧光素酶报告基因分析鉴定ER+和HER2+的BT474细胞和ER-和HER2+的SKBR3乳腺癌细胞作为对比研究对象；在E2存在与否的情况下，采用细胞增殖分析比较了Heregulin诱导的BT474和SKBR3细胞对Herceptin敏感性的改变；采用Western blot检测了HER2途径下游关键分子MAPK的磷酸化水平。结果在没有E2存在的情况下，Herceptin可有效抑制BT474和SKBR3细胞的增殖；在E2存在的情况下，Herceptin对BT474细胞增殖的抑制效应消失，同时，Herceptin不能降低MAPK的磷酸化；这些现象在SKBR3乳腺癌细胞中却不会出现。结论E2可通过ER非基因组活性影响ER+的乳腺癌细胞Herceptin的治疗效果，并可能参与了Herceptin抵抗的发生。 关键词Herceptin；乳腺癌；抵抗；ER；非基因组活性中图分类号文献标识码 A The effect and mechanism of estrogen and its receptor on the sensitivity of Herceptin in ER positive breast cancer through non-genomic activity Jian Xiao-Mei， Chen Bin， Wang Yue， Wang Yang， Li Yuping， Lou Guiyu， Zhang Yan， He FengtiaDepartment of Biochemistry and Molecular Biology ， Third Military Medical University， Chongqing， 400038， ChinaAbstract] Objective To explore the effect and mechanism of estrogen and its receptor on the sensitivity of Herceptin in ER positive breast cancer through non-genomic activity. Methods ER-positive/HER2-positive BT474 and ER-negative/HER2-positive SKBR3 breast cancer cells was indentified as a comparable model using western blot， transient transfection and repoter assay. In the presence or obsence of estrogen，the sensitivity of Herceptin on inhibiting Hereguilin-induced cell proliferation was detected by cell proliferation assay and the effect of Herceptin on the levels of phosphorylation of MAPK，the key melecules downstream of Her2 pathway was observed by Western blot. Results In obsence of estrogen， Herceptin effectively inhibits Hereguilin-induced proliferation of BT474 and SKBR3 cells. In presence of estrogen， the inhibitory effect of herceptin on Hereguilin-induced BT474 cell proliferation is disappered and Herceptin could not inhibit Hereguilin-induced phosphorylation of MAPK. These changes could not be observed in SKBR3 cells. Conclusions E2