论文部分内容阅读
目的探讨新的促癌分子肿瘤坏死因子-α诱导蛋白8样蛋白3(TIPE3)是否影响乳腺癌细胞MCF-7对阿霉素的敏感性及其机制。方法以0、0.5、1.0、1.5μg/m L阿霉素作用于TIPE3表达质粒转染的MCF-7细胞,分别采用细胞活性检测试剂盒(CCK-8)、流式细胞术分析TIPE3对阿霉素作用的MCF-7细胞相对活性及细胞周期分布的影响;采用Annexin V/PI染色流式细胞术检测分析TIPE3对阿霉素作用的MCF-7细胞凋亡的影响;采用实时荧光定量PCR(qRT-PCR)法和蛋白印记(Western blotting)法检测核因子-kappa B(NF-κB)信号通路活化情况、多药耐药基因1(MDR 1)及多药耐药蛋白P-糖蛋白(P-gp)的表达。结果过表达TIPE3增加了阿霉素处理MCF-7细胞的相对存活率和G2/M期细胞数,降低细胞凋亡率;NF-κB信号通路活化水平增加,多药耐药基因MDR1及其表达蛋白P-gp的表达水平升高。结论 TIPE3通过活化NF-κB信号通路上调P-gp的表达进而降低MCF-7细胞对阿霉素的敏感性。
Objective To investigate whether TIPE3, a novel tumor suppressor molecule, affects the sensitivity of breast cancer cell line MCF-7 to doxorubicin and its mechanism. Methods MCF-7 cells transfected with TIPE3 expression plasmids were transfected with doxorubicin (0,0.5,1.0,1.5μg / ml). Cell viability assay kit (CCK-8) and flow cytometry The effect of TIPE3 on the apoptosis of MCF-7 cells induced by doxorubicin was analyzed by flow cytometry with Annexin V / PI staining. Real-time fluorescence quantitative PCR The activation of nuclear factor-kappa B (NF-κB) signaling pathway was detected by qRT-PCR and Western blotting. The multidrug resistance gene 1 (MDR 1) and multidrug resistance protein P-glycoprotein (P-gp) expression. Results Overexpression of TIPE3 increased the relative viability of adriamycin-treated MCF-7 cells and the number of cells in G2 / M phase, and decreased the apoptosis rate. The activation of NF-κB signaling pathway increased, the expression of multidrug resistance gene MDR1 and its expression The expression of protein P-gp is increased. Conclusion TIPE3 can up-regulate the expression of P-gp by activating NF-κB signaling pathway and thus reduce the sensitivity of MCF-7 cells to doxorubicin.