| 摘要: |
| 目的：探讨环状RNA circNHSL1在小儿神经母细胞瘤中的表达及化疗耐受中的作用。方法：收集2016-2019年于我院就诊并接受手术联合阿霉素和顺铂化疗的神经母细胞瘤患儿60例，其中化疗敏感患儿35例，化疗耐受患儿25例。采用实时荧光定量聚合酶链式反应（qPCR）检测circNHSL1在神经母细胞瘤和化疗耐受患儿中的表达差异。构建人神经母细胞瘤细胞SH-SY5Y的阿霉素耐药株，检测阿霉素耐药细胞的circNHSL1表达量。通过沉默circNHSL1，采用细胞计数试剂盒（CCK-8）和流式细胞仪观察细胞增殖和凋亡情况。采用酶联免疫吸附试验（ELISA）和蛋白免疫印记法检测可能的转化生长因子-β（TGF-β）/Smad信号通路改变，并给予人重组TGF-β1进行回复实验，观察其在circNHSL1沉默细胞中的作用。结果：环状RNA circNHSL1在神经母细胞瘤中的表达高于正常组织（P<0.01），在化疗耐受患儿和耐药细胞株中的表达高于化疗敏感患儿（P<0.05）。在化疗耐受的细胞中，沉默circNHSL1细胞增殖活性降低，而凋亡率升高（P<0.01），TGFβ1分泌水平和表达量及其下游p-Smad2、p-Smad3表达量均降低（P<0.01）。再次给予人重组TGF-β1处理circNHSL1沉默的耐药细胞后，细胞增殖活性升高，而凋亡率降低，恢复耐药特性。结论：环状RNA circNHSL1在小儿神经母细胞瘤和化疗耐受组织中存在高表达现象，可通过激活TGF-β/Smad信号通路参与化疗耐受作用。 |
| 关键词: 环状RNA circNHSL1 神经母细胞瘤 化疗耐受 |
| DOI：doi:10.13407/j.cnki.jpp.1672-108X.2022.09.001 |
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| 基金项目: |
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| Expression of Circular RNA circNHSL1 in Pediatric Neuroblastoma and Its Role in Chemotherapy Resistance |
| Liu Jingdong, Wang Jinju |
| (Qingdao Women and Children’s Hospital, Shandong Qingdao 266034, China) |
| Abstract: |
| Objective: To probe into the expression of circular RNA circNHSL1 in pediatric neuroblastoma and its role in chemotherapy resistance. Methods: A total of 60 children with neuroblastoma who received surgery combined with doxorubicin and cisplatin chemotherapy in our hospital from 2016 to 2019 were collected, including 35 cases with chemotherapy sensitivity and 25 cases with chemotherapy resistance. Quantitative real-time polymerase chain reaction (qPCR) was used to detect the expression difference of circNHSL1 in children with neuroblastoma and chemotherapy resistance. Adriamycin-resistant strain of human neuroblastoma cell SH-SY5Y was performed, and the expression of circNHSL1 in the adriamycin-resistant cells was detected. Through circNHSL1-silenced cells, cell proliferation and apoptosis were observed by cell counting kit (CCK-8) and flow cytometry. Enzyme-linked immunosorbent assay (ELISA) and western blotting were used to detect the possible changes in transforming growth factor-β (TGF-β)/Smad signaling pathway, and human recombinant TGF-β1 was administered for recovery experiment to observe its role in circNHSL1-silenced cells. Results: The expression of circular RNA circNHSL1 in neuroblastoma was higher than that in normal tissues (P<0.01), and the expression in chemotherapy-resistant children and drug-resistant cell strains was higher than that in chemotherapy-sensitive children (P<0.05). In chemotherapy-resistant cells, the proliferation activity of circNHSL1-silenced cells decreased, while the apoptosis rate increased (P<0.01), the secretion level and expression of TGF-β1 and its downstream expression of p-Smad2 and p-Smad3 decreased (P<0.01). After treated with human recombinant TGF-β1, the cell proliferation activity increased significantly, while the apoptosis rate decreased, and the drug resistance was restored. Conclusion: Circular RNA circNHSL1 is highly expressed in children with neuroblastoma and chemotherapy resistant tissues, which may participate into the chemotherapy resistance by activating TGF-β/Smad signaling pathway. |
| Key words: circular RNA circNHSL1 neuroblastoma chemotherapy resistance |
