ZIC1基因转染联合姜黄素对人乳腺癌细胞生物学行为的影响

［摘要］目的: 探讨ZIC1基因转染联合姜黄素对人乳腺癌MDA-MB-231细胞生物学行为的影响。方法: 将慢病毒载体pLV Zic1 PGK Puro稳定转染人乳腺癌MDA-MB-231细胞，通过蛋白质印迹法检测转染后MDA-MB-231细胞ZIC1蛋白的表达水平。通过MTT法检测抑制率在50％左右的姜黄素浓度，并作为后续实验的标准加药浓度。以ZIC1空载不加姜黄素为对照组，空载加姜黄素、转染不加姜黄素及转染加姜黄素为实验组，以MTT法检测ZIC1、姜黄素及ZIC1联合姜黄素对MDA-MB-231细胞黏附的影响；用划痕实验检测各组细胞的迁移能力；用流式细胞术检测各组细胞的凋亡率。结果： 乳腺癌细胞转染ZIC1质粒后，ZIC1蛋白呈阳性表达，而转染空载体细胞中ZIC1蛋白表达阴性。细胞增殖抑制率在50％左右的姜黄素浓度为5 mg/L，以此作为标准加药浓度。与对照组比较，转染组、姜黄素组及转染联合姜黄素组乳腺癌MDA-MB-231细胞增殖受到明显抑制，凋亡率明显增高(均P＜0.05)。其中，ZIC1转染联合姜黄素对细胞增殖的抑制及凋亡诱导作用更明显(P＜0.05)。结论： ZIC1基因与姜黄素对乳腺癌的抑制存在协同作用。

Influence of ZIC1 gene combined with curcumin on biological behavior of human breast cancer MDA MB 231 cell

Abstract]Objective: To investigate the influence of ZIC1 gene combined with curcumin on biological behavior of human breast cancer cell MDA MB 231. Methods: Lentivirus vector pLV-Zic1-PGK-Puro was constructed and transfected stably into MDA-MB 231 cell，Western blotting were applied to detect the expression of ZIC1 protein in stably transfected MDA-MB-231 cell.For the following experiments,MDA MB 231 cell with no ZIC1 empty vector that we adding curcumin was defined as control group,and the groups of empty vector with adding curcumin,ZIC1 vector with no curcumin,ZIC1 vector with adding curcumin were used as experiment group.MTT assay was used to detect the effect of curcumin(various concentration) on cell proliferation,the standard concentration of curcumin was used in cell adhesion experiment and cell migration experiment when inhibition ratio was 50%，the two experiments were applied to detect differences of cell adhesion rate and migration ability. Cell apoptosis was then tested by FCM assay. Results: Expression of ZIC1 protein were positive in experiment group.The curcumin concentration was 5 mg/L when inhibition ratio was 50%.Compared with control group,ZIC1,curcumin,combination with ZIC1 and curcumin groups showed markedly cell proliferation repression and apoptosis induction(both P＜0.05）.And then，compared with the groups of empty vector with adding curcumin,ZIC1 vector with no curcumin,the effect of experiment group（adding curcumin） for cell proliferation repression and apoptosis induction was more obvious(P＜0.05). Conclusion: The combination with transfection of ZIC1 gene and curcumin worked most effectively in breast caner MDA-MB-231 cell proliferation repression and apoptosis induction,so ZIC1 gene and curcumin maybe has synergistic effect on tumor suppressor.