通过沉默 STAT3、Notch-1 和 β-catenin 基因增强野生型和耐药 MDA-MB-231 三阴性乳腺癌细胞系对多柔比星的化疗敏感性。
Enhancing chemosensitivity of wild-type and drug-resistant MDA-MB-231 triple-negative breast cancer cell line to doxorubicin by silencing of STAT 3, Notch-1, and β-catenin genes.
机构信息
Cell Therapy Center, The University of Jordan, PO Box: 5825, Amman, 11942, Jordan.
Department of Pharmacology, Faculty of Medicine, The University of Jordan, Amman, 11942, Jordan.
出版信息
Breast Cancer. 2020 Sep;27(5):989-998. doi: 10.1007/s12282-020-01098-9. Epub 2020 Apr 23.
BACKGROUND/OBJECTIVE: The absence of receptors in triple-negative breast cancer limits therapeutic choices utilized in clinical management of the disease. Doxorubicin is an important member of therapeutic regimens that is hindered by emergence of resistance. The current work aim to investigate of therapeutic potential of single and combinations of siRNA molecules designed for silencing STAT 3, Notch-1, and β-catenin genes in wild type and doxorubicin resistant MDA-MB-231 triple negative breast cancer cell line.
METHODS
Doxorubicin resistant MDA-MB-231 cell line was developed and characterized for the expression of multidrug resistance-related genes, CD44/CD24 markers, inflammatory cytokines, and the expression of STAT 3, Notch-1, and β-catenin targeted genes. Further, the effect of single and combinations of siRNA on cell viability and chemosensitivity of both wild type MDA-MB-231 cells (MDA-MB-231/WT) and doxorubicin resistant MDA-MB-231 cells (MDA-MB-231/DR) were assessed by MTT assay.
RESULTS
The IC of doxorubicin was 10-folds higher in MDA-MB-231/DR resistant cells compared to MDA-MB-231/WT control cells, 1.53 ± 0.24 μM compared to 0.16 ± 0.02 μM, respectively. The expression of targeted genes was higher in resistant cells compared to control cells, 3.6 ± 0.16 folds increase in β-catenin, 2.7 ± 0.09 folds increase in Notch-1, and 1.8 ± 0.09 folds increase in STAT-3. Following treatment with siRNAs, there was a variable reduction in mRNA expression of each of the targeted genes compared to scrambled siRNA and a reduction in IC in both cell lines. The effect of a combination of three genes produced the largest reduction in IC in resistant cell line.
CONCLUSION
Our study showed that the silencing of single and multiple genes involved in drug resistance and tumor progression by siRNA can enhance the chemosensitivity of cancer cells to conventional chemotherapy.
背景/目的：三阴性乳腺癌中受体的缺失限制了疾病临床治疗管理中使用的治疗选择。阿霉素是治疗方案中的重要成员，但由于耐药性的出现而受到阻碍。本研究旨在研究针对 STAT3、Notch-1 和 β-catenin 基因的 siRNA 分子的单药和联合治疗在野生型和阿霉素耐药 MDA-MB-231 三阴性乳腺癌细胞系中的治疗潜力。
方法
开发并鉴定阿霉素耐药 MDA-MB-231 细胞系，以检测多药耐药相关基因、CD44/CD24 标志物、炎症细胞因子以及 STAT3、Notch-1 和 β-catenin 靶向基因的表达。进一步，通过 MTT 分析评估 siRNA 对野生型 MDA-MB-231 细胞（MDA-MB-231/WT）和阿霉素耐药 MDA-MB-231 细胞（MDA-MB-231/DR）的细胞活力和化疗敏感性的单药和联合作用。
结果
与 MDA-MB-231/WT 对照细胞相比，MDA-MB-231/DR 耐药细胞的阿霉素 IC 高 10 倍，分别为 1.53±0.24 μM 和 0.16±0.02 μM。与对照细胞相比，耐药细胞中靶向基因的表达更高，β-catenin 表达增加 3.6±0.16 倍，Notch-1 表达增加 2.7±0.09 倍，STAT-3 表达增加 1.8±0.09 倍。用 siRNA 处理后，与 scrambled siRNA 相比，每个靶向基因的 mRNA 表达均有不同程度的降低，并且在两种细胞系中 IC 均降低。三种基因联合治疗对耐药细胞系的 IC 降低最大。
结论
我们的研究表明，通过 siRNA 沉默耐药和肿瘤进展相关的单个和多个基因可以增强癌细胞对常规化疗的敏感性。
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