PO.ET01.04 · 实验与分子治疗
下调AURKA和AURKB通过调控SNAIL1的表达促进TNBC细胞侵袭的减少
Downregulation of AURKA and AURKB promotes a decrease in cell invasion in TNBC by modulating the expression of SNAIL1
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摘要 Abstract
中文摘要
尽管化疗仍是TNBC的金标准治疗，但患者常经历高复发率。此外，多项研究表明，与非西班牙裔白人女性（NHW）相比，非西班牙裔黑人（NHB）和西班牙裔/拉丁裔（H/L）女性更可能被诊断并死于TNBC，表明TNBC存在显著的健康差异。虽然不同的分子参与者可驱动TNBC的这种侵袭性和耐药性，但AURKA和AURKB等有丝分裂激酶在被诊断为TNBC的NHB中过表达。AURKA和AURKB通过促进中心体成熟、有丝分裂纺锤体形成以及正确的染色体排列和分离，在调控细胞周期中发挥关键作用。然而，当失调时，它们可通过促进EMT驱动因子的表达而促进上皮-间质转化（EMT）。我们假设AURKA和AURKB的联合敲低将显著降低EMT驱动因子的表达并减弱细胞的侵袭能力。TCGA数据提示AURKA和AURKB在乳腺癌中显著过表达。Kaplan-Meier分析表明，与表达正常的女性相比，AURKA和AURKB的过表达与更差的无复发生存率相关。在进行siRNA介导的敲低后，进行了MTT实验、免疫印迹、免疫荧光和侵袭实验。MTT数据表明，共同靶向AURKA和AURKB降低了细胞活力，但大多数细胞仍在增殖。免疫印迹数据提示，共同靶向AURKA和AURKB显著降低了MDA-MB-157（NHB）中SNAIL的表达，而免疫荧光数据表明两株细胞系中Beta-catenin表达均减少。此外，在两株细胞系中共同靶向AURKA和AURKB导致细胞侵袭能力显著降低。MDA-MB-231（NHW）的初步结果也提示单独敲低SNAIL1后侵袭能力下降。因此，这表明AURKA和AURKB可在TNBC中beta-catenin和SNAIL1之间形成特定的轴。作为未来方向，我们将联合使用化学抑制剂以评估这些EMT生物标志物的表达。此外，我们将在体内模型中研究肿瘤生长和转移率。总之，由于TNBC的治疗选择有限，我们的结果表明，联合靶向AURKA和AURKB可能是降低TNBC转移表型的潜在方法，同时降低每种药物的剂量，从而减少患者的毒性。
查看英文原文 English abstract
Although chemotherapy is still the gold standard treatment in TNBC, patients often experience high levels of recurrence. Additionally, several studies have demonstrated that non-Hispanic Black (NHB) and Hispanic/Latino (H/L) women are more likely to be diagnosed and die from TNBC compared to non-Hispanic White women (NHW), indicating a significant health disparity in TNBC. While different molecular players can drive this aggressiveness and resistance in TNBC, mitotic kinases such as AURKA and AURKB are overexpressed in NHB diagnosed with TNBC. AURKA and AURKB play a pivotal role in regulating the cell cycle by promoting centrosome maturation, mitotic spindle formation, and proper chromosome alignment and segregation. However, when dysregulated, they can contribute to the epithelial-to-mesenchymal transition (EMT) by promoting the expression of EMT drivers. We hypothesize that the combined knockdown of AURKA and AURKB will significantly decrease the expression of EMT drivers and reduce the cells' ability to invade. TCGA data suggests a significant overexpression of AURKA and AURKB in breast cancer. Kaplan-Meier analysis indicates that overexpression of AURKA and AURKB is associated with a worse relapse-free survival rate compared to women who have normal expression. After performing the siRNA-mediated knockdown, MTT assay, immunoblotting, immunofluorescence, and invasion assays were performed. MTT data demonstrate that co-targeting AURKA and AURKB decrease cell viability; but most cells still proliferate. Immunoblotting data suggests that co-targeting AURKA and AURKB significantly decreases SNAIL expression in MDA-MB-157 (NHB), while immunofluorescence data indicate a decrease in Beta-catenin expression in both cell lines. Moreover, co-targeting AURKA and AURKB in both cell lines results in a significant reduction in the cells' ability to invade. Preliminary results in MDA-MB-231 (NHW) also suggest a decrease in the invasion capacity after a single knockdown of SNAIL1. Thus, this indicates that AURKA and AURKB can form a specific axis between beta-catenin and SNAIL1 in TNBC. As a future direction, we will use chemical inhibitors in combination to evaluate the expression of these EMT biomarkers. In addition, we will study tumour growth and metastasis rates in vivo models. In conclusion, since treatment options are limited in TNBC, our results suggest that combined targeting of AURKA and AURKB may be a potential approach to decrease the metastatic phenotype of TNBC while lowering the dose of each drug and thus the toxicity in patients.
利益披露 Disclosure
J. A. Orengo Orengo, None.