Department of Translational Oncology, Institute of Health Sciences, Dokuz Eylul University, Izmir 35330, Turkey.
Department of Translational Oncology, Institute of Oncology, Dokuz Eylul University, Izmir 35330, Turkey.
Nutrients. 2023 Nov 7;15(22):4705. doi: 10.3390/nu15224705.
Epithelial-mesenchymal transition (EMT) plays an important role in the biological and biochemical processes of cells, and it is a critical process in the malignant transformation, and mobility of cancer. Additionally, EMT is one of the main mechanisms contributing to chemoresistance. Resistance to oxaliplatin (OXA) poses a momentous challenge in the chemotherapy of advanced colorectal cancer (CRC) patients, highlighting the need to reverse drug resistance and improve patient survival. In this study, we explored the response of cyanidin-3--glucoside (C3G), the most abundant anthocyanin in plants, on the mechanisms of drug resistance in cancer, with the purpose of overcoming acquired OXA resistance in CRC cell lines.
We generated an acquired OXA-resistant cell line, named HCT-116-ROx, by gradually exposing parental HCT-116 cells to increasing concentrations of OXA. To characterize the resistance, we performed cytotoxicity assays and shape factor analyses. The apoptotic rate of both resistant and parental cells was determined using Hoechst 33342/Propidium Iodide (PI) fluorescence staining. Migration capacity was evaluated using a wound-healing assay. The mesenchymal phenotype was assessed through qRT-PCR and immunofluorescence staining, employing E-cadherin, N-cadherin, and Vimentin markers.
Resistance characterization announced decreased OXA sensitivity in resistant cells compared to parental cells. Moreover, the resistant cells exhibited a spindle cell morphology, indicative of the mesenchymal phenotype. Combined treatment of C3G and OXA resulted in an augmented apoptotic rate in the resistant cells. The migration capacity of resistant cells was higher than parental cells, while treatment with C3G decreased the migration rate of HCT-116-ROx cells. Analysis of EMT markers showed that HCT-116-ROx cells exhibited loss of the epithelial phenotype (E-cadherin) and gain of the mesenchymal phenotype (N-cadherin and Vimentin) compared to HCT-116 cells. However, treatment of resistant cells with C3G reversed the mesenchymal phenotype.
The morphological observations of cells acquiring oxaliplatin resistance indicated the loss of the epithelial phenotype and the acquisition of the mesenchymal phenotype. These findings suggest that EMT may contribute to acquired OXA resistance in CRC. Furthermore, C3G decreased the mobility of resistant cells, and reversed the EMT process, indicating its potential to overcome acquired OXA resistance.
上皮-间充质转化（EMT）在细胞的生物学和生物化学过程中发挥着重要作用，是癌症恶性转化和迁移的关键过程。此外，EMT 是导致化疗耐药的主要机制之一。奥沙利铂（OXA）耐药是晚期结直肠癌（CRC）患者化疗的重大挑战，这凸显了逆转耐药和提高患者生存的必要性。在这项研究中，我们探讨了植物中最丰富的花色苷——矢车菊素-3-葡萄糖苷（C3G）对癌症耐药机制的反应，旨在克服 CRC 细胞系获得性 OXA 耐药。
我们通过逐渐将亲本 HCT-116 细胞暴露于递增浓度的 OXA 中来产生获得性 OXA 耐药细胞系，命名为 HCT-116-ROx。为了表征耐药性，我们进行了细胞毒性测定和形态因子分析。使用 Hoechst 33342/碘化丙啶（PI）荧光染色测定两种耐药和亲本细胞的凋亡率。使用划痕愈合试验评估迁移能力。通过 qRT-PCR 和免疫荧光染色评估间充质表型，使用 E-钙粘蛋白、N-钙粘蛋白和波形蛋白标志物。
耐药特征表明，与亲本细胞相比，耐药细胞对 OXA 的敏感性降低。此外，耐药细胞呈现出纺锤形细胞形态，提示存在间充质表型。C3G 和 OXA 的联合治疗导致耐药细胞的凋亡率增加。耐药细胞的迁移能力高于亲本细胞，而 C3G 处理降低了 HCT-116-ROx 细胞的迁移率。EMT 标志物分析表明，与 HCT-116 细胞相比，HCT-116-ROx 细胞表现出上皮表型（E-钙粘蛋白）的丧失和间充质表型（N-钙粘蛋白和波形蛋白）的获得。然而，C3G 处理耐药细胞逆转了间充质表型。
细胞获得奥沙利铂耐药的形态学观察表明，上皮表型的丧失和间充质表型的获得。这些发现表明 EMT 可能有助于 CRC 中获得性 OXA 耐药。此外，C3G 降低了耐药细胞的迁移能力，并逆转了 EMT 过程，表明其有潜力克服获得性 OXA 耐药。
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