在顺铂耐药的卵巢癌细胞系中，KRT14基因敲低通过降低LRP11表达水平来降低顺铂耐药性。
KRT14 knockdown reduces cisplatin resistance by lowering LRP11 expression levels in cisplatin-resistant ovarian cancer cell lines.
作者信息
Liu Zequn, Weng Tingsong, Cheng Mi, Lei Tingying, Xiao Du, Deng Qiong, Wu Tianmei
机构信息
Department of Obstetrics, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, China.
Department of Prenatal Diagnostic Center, Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou, China.
出版信息
Transl Cancer Res. 2025 Mar 30;14(3):1786-1798. doi: 10.21037/tcr-24-1795. Epub 2025 Mar 14.
BACKGROUND
Platinum resistance is a major cause of mortality in patients with advanced ovarian cancer. Understanding the mechanisms underlying this resistance is essential for developing effective treatments to improve patient survival. Therefore, this study aimed to explore the role and mechanisms of keratin 14 (KRT14) in regulating cisplatin resistance in ovarian cancer.
METHODS
We utilized quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and western blotting to measure messenger RNA (mRNA) and protein expression levels, respectively. Cisplatin-resistant cell lines (SK-OV-3/DDP and A2780/DDP) were transfected with small interfering RNA (siRNA) targeting KRT14 (si-KRT14) or a plasmid containing low-density lipoprotein receptor-related protein 11 (LRP11) to knock down KRT14 or overexpress LRP11, respectively. Differentially expressed mRNAs were identified using Illumina RNA sequencing. Cell viability and half-maximal inhibitory concentration (IC) values were determined via cell counting kit-8 (CCK-8) assays, while apoptosis was assessed using flow cytometry and Hoechst 33258 staining.
RESULTS
KRT14 mRNA and protein levels were significantly higher in SK-OV-3/DDP and A2780/DDP cells compared with their parental counterparts. KRT14 knockdown reduced the IC values, increased apoptosis, and decreased the levels of the multidrug resistance (MDR)-related proteins P-glycoprotein (P-gp) and MDR-associated protein 1 (MRP1). KRT14 knockdown in SK-OV-3/DDP and A2780/DDP cells revealed 24 differentially expressed mRNAs. Further analysis revealed that KRT14 knockdown notably reduced LRP11 expression. LRP11 overexpression increased IC values, suppressed apoptosis, and enhanced MDR-related protein expression, thus counteracting the effects of KRT14 knockdown.
CONCLUSIONS
Cisplatin-resistant ovarian cancer cell lines revealed elevated KRT14 expression. KRT14 knockdown reduced cisplatin resistance by lowering LRP11 expression. Therefore, KRT14 may play a crucial role in mediating cisplatin resistance in ovarian cancer.
背景
铂耐药是晚期卵巢癌患者死亡的主要原因。了解这种耐药的潜在机制对于开发有效的治疗方法以提高患者生存率至关重要。因此，本研究旨在探讨角蛋白14（KRT14）在调节卵巢癌顺铂耐药中的作用及机制。
方法
我们分别利用定量逆转录-聚合酶链反应（qRT-PCR）和蛋白质印迹法来检测信使核糖核酸（mRNA）和蛋白质表达水平。用靶向KRT14的小干扰RNA（siRNA）（si-KRT14）或含低密度脂蛋白受体相关蛋白11（LRP11）的质粒转染顺铂耐药细胞系（SK-OV-3/DDP和A2780/DDP），分别敲低KRT14或过表达LRP11。使用Illumina RNA测序鉴定差异表达的mRNA。通过细胞计数试剂盒-8（CCK-8）检测法测定细胞活力和半数最大抑制浓度（IC）值，同时使用流式细胞术和Hoechst 33258染色评估细胞凋亡。
结果
与亲本细胞相比，SK-OV-3/DDP和A2780/DDP细胞中KRT14 mRNA和蛋白质水平显著更高。敲低KRT14降低了IC值，增加了细胞凋亡，并降低了多药耐药（MDR）相关蛋白P-糖蛋白（P-gp）和多药耐药相关蛋白1（MRP1）的水平。SK-OV-3/DDP和A2780/DDP细胞中敲低KRT14显示出24种差异表达的mRNA。进一步分析显示，敲低KRT14显著降低了LRP11表达。过表达LRP11增加了IC值，抑制了细胞凋亡，并增强了MDR相关蛋白表达，从而抵消了敲低KRT14的作用。
结论
顺铂耐药的卵巢癌细胞系显示KRT14表达升高。敲低KRT14通过降低LRP11表达降低了顺铂耐药性。因此，KRT14可能在介导卵巢癌顺铂耐药中起关键作用。
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