目的 基于转录组测序技术探讨隐丹参酮（cryptotanshinone，CPT）联合顺铂（cisplatin，DDP）干预对A2780/DDP细胞的影响及其潜在作用机制。方法 体外培养人卵巢癌A2780和A2780/DDP细胞，将细胞分为对照组、DDP组、DDP＋CPT给药组，采用MTS细胞增殖与细胞毒性检测试剂盒检测细胞存活率、划痕实验检测迁移能力、Transwell实验检测迁移和侵袭能力，探究DDP＋CPT干预下对A2780/DDP细胞的影响。获取各组的细胞mRNA进行转录组测序，筛选差异表达基因，拓扑分析得到CPT改善A2780/DDP对DDP耐药的核心靶点，对差异表达基因进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）富集分析，采用分子对接技术研究CPT与核心靶点的结合能力，实时荧光定量反转录聚合酶链式反应（real-time reverse transcription quantitative polymerase chain reaction，RT-qPCR）法检测核心靶点的mRNA表达情况。结果 A2780/DDP细胞的耐药指数为4；DDP＋CPT干预可显著抑制A2780/DDP耐药株的细胞存活率、细胞迁移和细胞侵袭能力（P＜0.001）。差异表达基因筛选结果表明，给药组与模型组相比共有253个差异表达基因，其中JUN原癌基因（JUN proto-oncogene gene，JUN）、Toll样受体2（Toll-like receptor 2，TLR2）、醌氧化还原酶1（quinone oxidoreductase 1，NQO1）、一氧化氮合酶2（nitric oxide synthase 2，NOS2）显著上调（P＜0.05），而趋化因子受体4（chemokine receptor 4，CXCR4）显著下降（P＜0.05）。GO和KEGG结果表明，CPT可能影响TLR信号通路、B细胞受体信号通路、白细胞介素-17等信号通路，从而改善A2780/DDP细胞对DDP的耐药性；RT-qPCR结果表明，CPT可上调JUN、TLR2、NOS2、NQO1的mRNA表达（P＜0.001），抑制CXCR4的mRNA表达（P＜0.001），改善A2780/DDP对DDP的耐药性。结论 CPT可改善A2780/DDP耐药细胞对DDP的耐药性，其机制可能与CPT上调JUN、TLR2、NOS2、NQO1的mRNA表达，下调CXCR4的mRNA表达有关。

Objective To investigated the effects of cryptotanshinone (CPT) combined with cisplatin (DDP) on A2780/DDP cells and its potential mechanism of action based on transcriptome sequencing technology.Methods The A2780 and A2780/DDP cells were cultured in vitro, and the cells were divided into control group, DDP group, and DDP + CPT experimental group. MTS cell proliferation and cytotoxicity detection Kit were used to detect the cell survival rate, scratch assay was used to detect the migratory ability, and Transwell assay was used to detect the migratory and invasive ability, to investigate the effect of DDP + CPT intervention on A2780/DDP cells. The mRNA of the above grouped cells was obtained for transcriptome sequencing, screening for differentially expressed genes, topology analysis was used to obtain the core target of CPT to improve the resistance of A2780/DDP to DDP, the gene ontology (GO) function and the Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis were conducted for differentially expressed genes, molecular docking technology was used to study the binding ability of CPT to the core target and real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA expression of the core target. Results The resistance index of A2780/DDP cells was four, DDP + CPT intervention significantly inhibited the cell viability, cell migration and cell invasion ability of A2780/DDP-resistant strains (P < 0.001). The results of differentially expressed genes screening showed that there were 253 differentially expressed genes in the drug group as compared with the model group, among which JUN proto-oncogene gene (JUN), Toll-like receptor 2 (TLR2), quinone oxidoreductase 1 (NQO1), and nitric oxide synthase 2 (NOS2) were significantly up-regulated (P < 0.05), while chemokine receptor 4 (CXCR4) was significantly decreased (P < 0.05). The results of GO and KEGG suggested that CPT might affect the TLR signaling pathway, B-cell receptor signaling pathway, and interleukin-17 signaling pathway and other mechanisms, thereby improving the drug resistance of A2780/DDP cells to DDP. RT-qPCR results showed that CPT up-regulated the mRNA expression of JUN, TLR2, NOS2, NQO1 (P < 0.001), inhibited CXCR4 mRNA expression (P < 0.001) to improve the drug resistance of A2780/DDP to DDP. Conclusion CPT could improve the resistance of A2780/DDP-resistant cells to DDP, and the mechanism may be related to the up-regulation of the mRNA expression of JUN, TLR2, NOS2, NQO1 and the down-regulation of CXCR4 mRNA expression by CPT.

R285.5

宁德市自然科学基金联合项目（2022J02）