Department of Pharmacy, University of Bonn, 53113 Bonn, Germany.
Department of RNA Metabolism, Institute of Bioorganic Chemistry Polish Academy of Sciences, 61-704 Poznań, Poland.
Int J Mol Sci. 2020 Dec 3;21(23):9240. doi: 10.3390/ijms21239240.
The microenvironment possesses a strong impact on the tumor chemoresistance when cells bind to components of the extracellular matrix. Here we elucidate the signaling pathways of cisplatin resistance in W1 ovarian cancer cells binding to collagen type 1 (COL1) and signaling interference with constitutive cisplatin resistance in W1CR cells to discover the targets for sensitization. Proteome kinase arrays and Western blots were used to identify the signaling components, their impact on cisplatin resistance was evaluated by inhibitory or knockdown approaches. W1 cell binding to COL1 upregulates integrin-associated signals via FAK/PRAS40/mTOR, confirmed by β1-integrin (ITGB1) knockdown. mTOR appears as key for resistance, its blockade reversed COL1 effects on W1 cell resistance completely. W1CR cells compensate ITGB1-knockdown by upregulation of discoidin domain receptor 1 (DDR1) as alternative COL1 sensor. COL1 binding via DDR1 activates the MAPK pathway, of which JNK1/2 appears critical for COL1-mediated resistance. JNK1/2 inhibition inverts COL1 effects in W1CR cells, whereas intrinsic cisplatin resistance remained unaffected. Remarkably, knockdown of HSP27, another downstream MAPK pathway component overcomes intrinsic resistance completely sensitizing W1CR cells to the level of W1 cells for cisplatin cytotoxicity. Our data confirm the independent regulation of matrix-induced and intrinsic chemoresistance in W1 ovarian cancer cells and offer novel targets for sensitization.
当细胞与细胞外基质的成分结合时，微环境对肿瘤的化疗耐药性有很强的影响。在这里，我们阐明了与胶原蛋白 I 型（COL1）结合的 W1 卵巢癌细胞中顺铂耐药的信号通路，并通过信号干扰发现了 W1CR 细胞中顺铂耐药的靶点，以实现增敏作用。采用蛋白激酶阵列和 Western blot 来鉴定信号成分，通过抑制或敲低方法评估它们对顺铂耐药性的影响。W1 细胞与 COL1 结合通过 FAK/PRAS40/mTOR 上调整合素相关信号，这一过程通过 β1 整合素（ITGB1）敲低得到证实。mTOR 似乎是耐药的关键，其阻断完全逆转了 COL1 对 W1 细胞耐药性的影响。W1CR 细胞通过上调盘状结构域受体 1（DDR1）作为 COL1 的替代传感器来补偿 ITGB1 敲低。DDR1 通过 COL1 结合激活 MAPK 通路，其中 JNK1/2 似乎对 COL1 介导的耐药性至关重要。JNK1/2 抑制可反转 COL1 在 W1CR 细胞中的作用，而内在的顺铂耐药性不受影响。值得注意的是，另一个下游 MAPK 通路成分 HSP27 的敲低完全克服了内在耐药性，使 W1CR 细胞对顺铂细胞毒性的敏感性提高到 W1 细胞的水平。我们的数据证实了 W1 卵巢癌细胞中基质诱导的和内在的化疗耐药性的独立调节，并为增敏提供了新的靶点。
