Cancer Research Institute, Hangzhou Cancer Hospital.
Department of Clinical Laboratory, Hangzhou Cancer Hospital, Hangzhou, Zhejiang Province.
Anticancer Drugs. 2020 Sep;31(8):776-784. doi: 10.1097/CAD.0000000000000952.
Cisplatin is a widely used chemotherapeutic drug in lung cancer treatment. Most cancer patients eventually develop cisplatin resistance, resulting in a poor prognosis. Previously, we identified a novel marker, family with sequence similarity 60A (FAM60A), that was responsible for resistance in cisplatin-resistant human lung adenocarcinoma A549 (A549/DDP) cells. Here, we investigated the biological effects of FAM60A in A549/DDP cells and explored the underlying molecular mechanisms to understand its functional role in cisplatin resistance. Real-time quantitative PCR and western blot analysis were used to determine the expression levels of FAM60A in A549/DDP cells. FAM60A and SKP2 were knockdown with small-interfering RNA (siRNA). Cancer cell viability was analyzed with flow cytometry. The mRNA and protein expression levels of FAM60A increased significantly and dose-dependently in A549/DDP cells following cisplatin treatment. FAM60A overexpression up-regulated MDR1 expression, inhibited caspase 3, cleaved-caspase 3, and caspase 8 expression, and prevented cancer cell death. Microarray analysis of cells transfected with siRNA against the FAM60A transcript and control samples showed that SKP2 expression was positively regulated by FAM60A. SKP2 knockdown using a short-hairpin RNA reversed the functions induced by FAM60A. These results suggest that overexpression of FAM60A in A549/DDP cells led to SKP2 upregulation and enhanced cisplatin resistance in cancer cells. These provide new insights into chemoresistance and may contribute to reversing cisplatin resistance during lung cancer treatment.
顺铂是肺癌治疗中广泛使用的化疗药物。大多数癌症患者最终会产生顺铂耐药性，导致预后不良。先前，我们鉴定了一种新型标志物，家族与序列相似性 60A（FAM60A），它负责顺铂耐药的人肺腺癌细胞 A549（A549/DDP）的耐药性。在这里，我们研究了 FAM60A 在 A549/DDP 细胞中的生物学效应，并探讨了潜在的分子机制，以了解其在顺铂耐药中的功能作用。实时定量 PCR 和 Western blot 分析用于确定 FAM60A 在 A549/DDP 细胞中的表达水平。用小干扰 RNA（siRNA）敲低 FAM60A 和 SKP2。用流式细胞术分析癌细胞活力。顺铂处理后，A549/DDP 细胞中 FAM60A 的 mRNA 和蛋白表达水平显著且剂量依赖性增加。FAM60A 过表达上调 MDR1 表达，抑制 caspase 3、cleaved-caspase 3 和 caspase 8 的表达，并阻止癌细胞死亡。用针对 FAM60A 转录本的 siRNA 转染的细胞和对照样品的微阵列分析显示，SKP2 的表达受 FAM60A 的正向调节。用短发夹 RNA 敲低 SKP2 逆转了 FAM60A 诱导的功能。这些结果表明，A549/DDP 细胞中 FAM60A 的过表达导致 SKP2 的上调，并增强了癌细胞对顺铂的耐药性。这些为化疗耐药性提供了新的见解，并可能有助于在肺癌治疗中逆转顺铂耐药性。
