Lu Yu, Zhao Xianyang, Yuan Mingliang, Zhao Ming, Liu Kaisheng, Zhang Miaomiao, Qiu Xiaoyan, Yu Xuechun, Liu Xinliang, Wei Dongping, Xie Jun, Cheng Zhongbin
Department of Clinical Research Center, Xuyi People's Hospital, Affiliated Xuyi Hospital of Yangzhou University Medical College, Jiangsu, China.
Department of pharmacy, Xuyi People's Hospital, Affiliated Xuyi Hospital of Yangzhou University Medical College, Jiangsu, China.
J Cancer. 2024 Mar 11;15(9):2475-2485. doi: 10.7150/jca.93229. eCollection 2024.
Chemotherapy resistance is a barrier to effective cancer prognoses. Cisplatin (CDDP) resistance is a major challenge for esophageal cancer (EC) therapy. A deeper understanding of the fundamental mechanisms of cisplatin resistance and improved targeting strategies are required in clinical settings. This study was performed to identify and characterize a marker of cisplatin resistance in EC cells. KYSE140 and Eca-109 cells were subjected to escalating concentrations of cisplatin, resulting in the development of cisplatin-resistant KYSE140/CDDP and Eca-109/CDDP cell lines, respectively. RNA Sequencing (RNA-seq) was utilized to screen for the genes exhibiting differential expression between cisplatin-resistant and parental cells. Reverse transcription quantitative PCR was conducted to assess gene expression, and western blotting was employed to analyze protein levels. A sphere-formation assay was performed to validate tumor cell stemness. Cell counting kit-8 (CCK-8) experiments were conducted to confirm the sensitivity of cells to cisplatin. We examined the relationship between target genes and the clinicopathological features of patients with EC. Furthermore, the expression of target genes in EC tissues was evaluated via western blotting and fluorescence probe hybridization (FISH). KYNU was upregulated in cisplatin-resistant EC cells (KYSE140/CDDP and Eca-109/CDDP cells) and in EC tissues compared to that in the respective parental cell lines (KYSE140 and Eca-109 cells) and non-carcinoma tissues. Downregulation of KYNU increased cell sensitivity to cisplatin and suppressed tumor stemness, whereas abnormal KYNU expression had the opposite effect. KYNU expression was correlated with the expression of tumor stemness-associated factors (SOX2, Nanog, and OCT4) and the tumor size. KYNU may promote drug resistance in EC by regulating cancer stemness, and could serve as a biomarker and therapeutic target for EC.
化疗耐药是有效癌症预后的一个障碍。顺铂（CDDP）耐药是食管癌（EC）治疗的一项重大挑战。临床环境中需要更深入地了解顺铂耐药的基本机制并改进靶向策略。本研究旨在鉴定和表征EC细胞中顺铂耐药的标志物。将KYSE140和Eca - 109细胞暴露于递增浓度的顺铂中，分别产生了顺铂耐药的KYSE140/CDDP和Eca - 109/CDDP细胞系。利用RNA测序（RNA-seq）筛选在顺铂耐药细胞和亲本细胞之间表现出差异表达的基因。进行逆转录定量PCR以评估基因表达，并采用蛋白质印迹法分析蛋白质水平。进行成球试验以验证肿瘤细胞干性。进行细胞计数试剂盒 - 8（CCK - 8）实验以确认细胞对顺铂的敏感性。我们研究了靶基因与EC患者临床病理特征之间的关系。此外，通过蛋白质印迹法和荧光探针杂交（FISH）评估EC组织中靶基因的表达。与各自的亲本细胞系（KYSE140和Eca - 109细胞）及非癌组织相比，KYNU在顺铂耐药的EC细胞（KYSE140/CDDP和Eca - 109/CDDP细胞）及EC组织中上调。KYNU的下调增加了细胞对顺铂的敏感性并抑制了肿瘤干性，而KYNU的异常表达则产生相反的效果。KYNU表达与肿瘤干性相关因子（SOX2、Nanog和OCT4）的表达及肿瘤大小相关。KYNU可能通过调节癌症干性促进EC中的耐药性，并可作为EC的生物标志物和治疗靶点。
