Li Lei, Yu Libo, Cao Xiansheng, Zhang Chao, Liu Qi, Chen Jun
Department of Gastrointestinal Surgery, Yantai Affiliated Hospital of Binzhou Medical University, Yantai, China.
Department of Radiology, Yantaishan Hospital, Yantai, China.
J Oncol. 2022 Apr 25;2022:3605436. doi: 10.1155/2022/3605436. eCollection 2022.
To further explore the mechanisms of curcumin reversing multidrug resistance (MDR) in HCT8/VCR cells. Here, we employed comparative analysis of proteomic of essential proteins of human colon carcinoma HCT8/VCR cells with or without treatment of curcumin by separating and quantifying the essential protein posttranslational modification through radical-free two-dimensional polyacrylamide gel electrophoresis with strong reductant. The reverse impact of curcumin on multidrug resistance of HCT8/VCR and HCT8/VCR cells was evaluated using MTT assay. After adding curcumin 25 M for 72 h, by 2-DE and mass spectrometry, twenty proteins were certified with changed expression levels. Three protein sites were upregulated and seventeen protein sites were downregulated in curcumin-treated HCT-8/VCR. Verification analyses were conducted using RT-PCR and Western blotting for downregulated proteins including GSTP1 and PRDX6. The proteins might have a direct or indirect contact with multidrug resistance. The finding of the research would provide novel sights for systematically comprehending the mechanisms of the reversal impacts of curcumin on MDR in HCT8/VCR cells and contribute to the recognition and application of new markers in clinical practice.
为进一步探究姜黄素逆转人结肠癌HCT8/VCR细胞多药耐药（MDR）的机制。在此，我们通过使用强还原剂的无自由基二维聚丙烯酰胺凝胶电泳分离和定量必需蛋白的翻译后修饰，对经姜黄素处理和未经处理的人结肠癌HCT8/VCR细胞的必需蛋白进行蛋白质组学比较分析。使用MTT法评估姜黄素对HCT8/VCR细胞多药耐药的逆转作用。加入25μM姜黄素处理72小时后，通过二维电泳和质谱分析，鉴定出20种表达水平发生变化的蛋白质。在经姜黄素处理的HCT-8/VCR细胞中，3个蛋白位点上调，17个蛋白位点下调。对包括GSTP1和PRDX6在内的下调蛋白进行RT-PCR和蛋白质印迹验证分析。这些蛋白质可能与多药耐药有直接或间接联系。该研究结果将为系统理解姜黄素对HCT8/VCR细胞多药耐药逆转作用的机制提供新的视角，并有助于在临床实践中识别和应用新的标志物。
