同源异型核基因1诱导乳腺癌细胞发生上皮-间充质转化对三苯氧胺耐药的研究  

Paired related homoeoboxl - induced breast cancer epithelial cell epithelial - mesenchymal transi-tion to tamoxifen resistance

作　　者：董娇娇 吕志栋[2] 李福年[2] 姜丹丹[2] 杨召川[3] Dong Jiaojiao;Lyu Zhidong;Li Funian;Jiang Dandan;Yang Zhaochuan(Medical School of Qingclao University, Oingdao 266021, China;Department of Breast Disease Diaglmsis and Treatment Centre, the Qingdao University Affiliated Hospital, Qingdao 266003, China;Department of Child Healtheare, the Qingdao University Affiliated Hospital, Qingdao 266003, China)

机构地区：[1]青岛大学医学部,266021 [2]青岛大学附属医院乳腺病诊疗中心,266003 [3]青岛大学附属医院儿童保健科,266003

出　　处：《中华实验外科杂志》2018年第6期1002-1004,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金（81302290、81700029）;山东省自然科学基金（ZR2017PH032）

摘　　要：目的观察同源异型核基因1（Prrxl）过表达诱导乳腺癌细胞发生上皮一间充质转化（EMT）对三苯氧胺（TAM）耐药的影响。方法脂质体转染法转染Pixxl低表达细胞株MCF-7细胞，实验组细胞为转染Prrxl基因的MCF-7细胞，空白对照组为转染空白载体的MCF-7细胞，阴性对照组为MCF-7细胞，倒置荧光显微镜观察转染效率，Westernblot验证转染成功，反转录一聚合酶链反应（RT-PCR）检测Prrxl对E-钙黏蛋白（E-cadherin，E-cad）及波形蛋白（Vimentin）mRNA表达水平的影响后，采用细胞计数试剂盒（CCK-8）检测3组细胞对i苯氧胺的耐药性。结果转染后荧光效率达80％以上，实验组Prrxl蛋白表达水平明显高于对照组，且差异有统计学意义（P=0．000），两对照组之间差异无统计学意义（P=0．326）。PrrxlmRNA在阴性对照组、空白对照组、实验组中CT值分别为13．502±0．360、13．770±0．642、5．203±0．231，实验组PrrxlmRNA表达水平为阴性对照组的320倍；E-cadmRNA在阴性对照组、空白对照组、实验组CT值分别为18．471±0．076、18．611±0．108、19．3944-0．400，实验组E-cadmRNA表达水平为阴性对照组的0．560倍；VimentinmRNA在阴性对照组、空白对照组、实验组CT值分别为19．4224-0．082、19．484±0．021、17．817±0．061，实验组VimentinmRNA表达水平为阴性对照组的3．030倍。三苯氧胺作用24h后，实验组半数抑制浓度（IC50）明显高于阴性对照组（P=0．000）和空白对照组（P=0．000），差异有统计学意义，而阴性对照组和空白对照组之间差异无统计学意义（P=0．150）。结论Prrxl过表达可诱导MCF-7细胞发生EMT现象促进对三苯氧胺产生耐药。Objective To investigate the effect of paired related homoeoboxl （Prrxl） on epithelial - mesenchymal transition （EMT） and the resistant ability to tamoxifen in breast cancer epithelial cells. Methods The Prrxl low expression cell line MCF -7 cells were transfected by liposome transfection, experimental group were MCF- 7 cells transfeeted with Prrxl gene, blank control group was MCF -7 cells transfected with black vector, negative control group was MCF -7 cells and the transfeetion was verified by Western blotting. The expression level of Prrxl , E - cadherin and Vimentin was detected by real - time quantitative polymerase chain reaction （ Real - time PCR ）. Cytotoxicity was detected by cell counting kit - 8 （ CCK - 8 ） after exposure to tamoxifen. Results After MCF - 7 cells were transfeeted with Prrxl, the fluorescence efficiency reached more than 80%. Western blotting results showed that the expression of Prrxl in experimental group was significantly higher than in control groups （ P = 0. 000 ） , and there was no no statistically significant difference between two control groups （ P=0. 326 ）. Real - time PCR results showed that the CT values of Prrxl mRNA in negative control, blank control and experimental groups were 13. 502 ± 0. 360, 13. 770±0. 642, 5. 203±0. 231 respectively. The Prrxl mRNA expression level in the experimen- tal group was 320 times that of the negative control group. The CT values of E - adherin mRNA in negative eontrnl, blank control and experimental groups were 18. 471± 0. 076, 18. 611 ± 0. 108, 19. 394 ± 0. 400 respectively. The E - cad mRNA expression level in the experimental group was 0. 560 times that of the negative control group. The CT values of vimentin mRNA in negative control, blank control and experimen- tal groups were 19. 422 ± 0. 082, 19. 484±0. 021,17. 817±0. 061 respectively. The Vimentin mRNA expression level in the experimental group was 3.03 times that of the negative control group. CCK - 8 results showed that after the action of tamoxif

关 键 词：乳腺癌 同源异型核基因1 上皮-间充质转化 三苯氧胺 耐药 

分 类 号：R737.9[医药卫生—肿瘤]