目的 基于轉(zhuǎn)錄組測序技術(shù)探討隱丹參酮（cryptotanshinone，CPT）聯(lián)合順鉑（cisplatin，DDP）干預(yù)對A2780/DDP細胞的影響及其潛在作用機制。方法 體外培養(yǎng)人卵巢癌A2780和A2780/DDP細胞，將細胞分為對照組、DDP組、DDP＋CPT給藥組，采用MTS細胞增殖與細胞毒性檢測試劑盒檢測細胞存活率、劃痕實驗檢測遷移能力、Transwell實驗檢測遷移和侵襲能力，探究DDP＋CPT干預(yù)下對A2780/DDP細胞的影響。獲取各組的細胞mRNA進行轉(zhuǎn)錄組測序，篩選差異表達基因，拓撲分析得到CPT改善A2780/DDP對DDP耐藥的核心靶點，對差異表達基因進行基因本體（gene ontology，GO）功能及京都基因與基因組百科全書（Kyoto encyclopedia of genes and genomes，KEGG）富集分析，采用分子對接技術(shù)研究CPT與核心靶點的結(jié)合能力，實時熒光定量反轉(zhuǎn)錄聚合酶鏈式反應(yīng)（real-time reverse transcription quantitative polymerase chain reaction，RT-qPCR）法檢測核心靶點的mRNA表達情況。結(jié)果 A2780/DDP細胞的耐藥指數(shù)為4；DDP＋CPT干預(yù)可顯著抑制A2780/DDP耐藥株的細胞存活率、細胞遷移和細胞侵襲能力（P＜0.001）。差異表達基因篩選結(jié)果表明，給藥組與模型組相比共有253個差異表達基因，其中JUN原癌基因（JUN proto-oncogene gene，JUN）、Toll樣受體2（Toll-like receptor 2，TLR2）、醌氧化還原酶1（quinone oxidoreductase 1，NQO1）、一氧化氮合酶2（nitric oxide synthase 2，NOS2）顯著上調(diào)（P＜0.05），而趨化因子受體4（chemokine receptor 4，CXCR4）顯著下降（P＜0.05）。GO和KEGG結(jié)果表明，CPT可能影響TLR信號通路、B細胞受體信號通路、白細胞介素-17等信號通路，從而改善A2780/DDP細胞對DDP的耐藥性；RT-qPCR結(jié)果表明，CPT可上調(diào)JUN、TLR2、NOS2、NQO1的mRNA表達（P＜0.001），抑制CXCR4的mRNA表達（P＜0.001），改善A2780/DDP對DDP的耐藥性。結(jié)論 CPT可改善A2780/DDP耐藥細胞對DDP的耐藥性，其機制可能與CPT上調(diào)JUN、TLR2、NOS2、NQO1的mRNA表達，下調(diào)CXCR4的mRNA表達有關(guān)。

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

寧德市自然科學基金聯(lián)合項目（2022J02）