白介素1受体相关激酶1在乳腺癌紫杉醇耐受患者治疗中的作用机制研究

发布时间：2019-05-18 21:14

【摘要】：目的探讨白介素1受体相关激酶1(IRAK1)在乳腺癌紫杉醇耐受患者治疗中的作用机制。方法选取2013年5月—2015年9月赣南医学院第一附属医院肿瘤科收集的乳腺组织(对照)及乳腺癌组织,其中对照女性16例,Basal-like型乳腺癌患者38例,Luminal-A型乳腺癌患者25例,Luminal-B型乳腺癌患者31例,HER2过表达型乳腺癌患者31例,三阴乳腺癌患者58例。取对照乳腺组织及乳腺癌组织,采用实时荧光定量PCR(q PCR)检测IRAK1 mRNA的相对表达水平,采用酶联免疫吸附试验(ELISA)法检测炎性因子白介素(IL)-6、IL-8和CXC趋化因子配体1(CXCL-1)表达水平。培养乳腺癌细胞株HMEC、MCF7、MB415、MB436、MB468、BT549、SUM159,采用体外低浓度梯度递增联合大剂量间断冲击方法诱导乳腺癌细胞株产生紫杉醇耐受性,检测未经处理的细胞株和紫杉醇耐受细胞株IRAK1 mRNA相对表达水平。经shRNA或空白序列NC-shRNA转染紫杉醇耐受乳腺癌细胞株,采用q PCR检测增殖相关基因细胞周期蛋白(Cyclin)D1、增殖细胞核抗原(PCNA)和Cyclin A相对表达水平,凋亡相关基因p53、c-myc、Bcl-2、c-erb-2相对表达水平,迁移相关基因Cullin 1、Bcl-6和KLF6相对表达水平,采用ALDEFLUOR实验检测醛脱氢酶(ALDH)活性。结果各类型乳腺癌紫杉醇耐受患者IRAK1 mRNA相对表达水平均高于不耐受患者(P0.05),三阴乳腺癌紫杉醇耐受患者IRAK1 mRNA相对表达水平均高于其他类型乳腺癌紫杉醇耐受患者(P0.05)。对照者及各类型乳腺癌紫杉醇耐受患者IL-6、IL-8、CXCL-1表达水平比较,差异均有统计学意义(P0.05)。紫杉醇耐受的各乳腺癌细胞株IRAK1 mRNA相对表达水平均高于未经紫杉醇处理的细胞株(P0.05)。经shRNA转染的紫杉醇耐受的各乳腺癌细胞株Cyclin D1、PCNA、Cyclin A相对表达水平均低于经NC-shRNA转染的各乳腺癌细胞株,p53、c-myc、Bcl-2、c-erb-2相对表达水平均高于经NC-shRNA转染的各乳腺癌细胞株,Cullin 1、Bcl-6相对表达水平、ALDH活性均低于经NC-shRNA转染的各乳腺癌细胞株,KLF6相对表达水平高于经NC-shRNA转染的各乳腺癌细胞株(P0.05)。结论 IRAK1在紫杉醇耐受的乳腺癌细胞,尤其是三阴乳腺癌细胞中发挥重要作用。抑制IRAK1表达可显著降低乳腺癌细胞的增殖和迁移能力,诱导癌细胞凋亡,IRAK1抑制剂可用于乳腺癌尤其是三阴乳腺癌的治疗。
[Abstract]:Objective to investigate the role of IL-1 receptor-associated kinase 1 (IRAK1) in the treatment of paclitaxel tolerance in breast cancer. Methods from May 2013 to September 2015, breast tissues and breast cancer tissues were collected from the Department of Oncology, the first affiliated Hospital of Gannan Medical College, including 16 women and 38 patients with Basal- like breast cancer. There were 25 patients with Luminal- A breast cancer, 31 patients with Luminal- B breast cancer, 31 patients with HER2 overexpression breast cancer and 58 patients with Sanyin breast cancer. The relative expression of IRAK1 mRNA was detected by real-time fluorescence quantitative PCR (q PCR) in control breast tissue and breast cancer tissue, and the inflammatory factor IL-6 was detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of IL-8 and CXC chemokine ligand 1 (CXCL-1). Cultured breast cancer cell line HMEC,MCF7,MB415,MB436,MB468,BT549,SUM159, was induced to produce paclitaxel tolerance by low concentration gradient increase in vitro combined with high dose intermittent shock. The relative expression of IRAK1 mRNA in untreated cell lines and paclitaxel tolerant cell lines was detected. Paclitaxel-tolerant breast cancer cell line was transformed into paclitaxel-tolerant breast cancer cell line by shRNA or blank sequence NC-shRNA. The relative expression of proliferation-related gene (Cyclin) D1, proliferating cell nuclear antigen (PCNA) and Cyclin A and apoptosis-related gene p53 were detected by Q-PCR. The relative expression levels of Cullin 1, Bcl 6 and KLF6 were detected by ALDEFLUOR assay. Results the relative expression level of IRAK1 mRNA in paclitaxel tolerant patients with all types of breast cancer was higher than that in patients with intolerable breast cancer (P 0.05). The relative expression of IRAK1 mRNA in patients with paclitaxel tolerance in Sanyin breast cancer was higher than that in patients with paclitaxel tolerance in other types of breast cancer (P 0.05). There was significant difference in the expression of IL-6,IL-8,CXCL-1 between the control group and the patients with paclitaxel tolerance in various types of breast cancer (P 0.05). The relative expression level of IRAK1 mRNA in paclitaxel tolerant breast cancer cell lines was higher than that without paclitaxel treatment (P 0.05). The relative expression levels of Cyclin A in paclitaxel-tolerant breast cancer cell lines Cyclin D1 and PCNA were lower than those in breast cancer cell lines p53, c and Bcl, which were transformed by NC-shRNA, and the relative expression levels of Cyclin A in paclitaxel-tolerant breast cancer cell lines p53, c and Bcl were lower than those in each breast cancer cell line, p53, c and Bcl. The relative expression level of c-erb-2 was higher than that of breast cancer cell lines transfected with NC-shRNA. The relative expression levels of Bcl-6 and ALDH in Cullin-1 and Bcl-6 cells were lower than those in breast cancer cell lines transfected with NC-shRNA. The relative expression level of KLF6 was higher than that of breast cancer cell lines transfected with NC-shRNA (P 0.05). Conclusion IRAK1 plays an important role in paclitaxel tolerant breast cancer cells, especially Sanyin breast cancer cells. Inhibition of IRAK1 expression can significantly reduce the proliferation and migration of breast cancer cells and induce apoptosis of breast cancer cells. IRAK1 inhibitor can be used in the treatment of breast cancer, especially Sanyin breast cancer.
【作者单位】：赣南医学院第一附属医院肿瘤科;
【分类号】：R737.9

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