| 耿硕,韩倩,孟琳琳,等.自分泌 IL-15 的 HER2-CAR-NK92细胞的构建及体外抗肿瘤活性研究[J].中国海洋药物,2025,44(5):51-56. |
| 自分泌 IL-15 的 HER2-CAR-NK92细胞的构建及体外抗肿瘤活性研究 |
| Construction and antitumor activity of HER2-CAR-NK92 cells secreting IL-15 |
| 投稿时间：2024-04-01 修订日期：2024-07-23 |
| DOI：10.13400/j.cnki.cjmd.2025.05.008 |
| 中文关键词: 嵌合抗原受体 NK-92 细胞 IL-15 HER2 免疫治疗 |
| English Keywords:chimeric antigen receptor NK-92 cell IL-15 HER2 immunotherapy |
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| 中文摘要: |
| 目的 探索自分泌白细胞介素 15(IL-15)对 NK-92 细胞的促增殖作用,以及靶向人表皮生长因子受体 2(HER2)的嵌合抗原受体(CAR)NK-92 细胞对相关肿瘤细胞的杀伤作用。方法 利用慢病毒载体将 IL-15 导入 NK-92 细胞,ELISA 检测 IL-15 的表达量,CCK-8 法检测其增殖活性,在此基础上导入靶向 HER2 的二代 CAR,流式细胞术检测 CAR 的阳性率,体外杀伤实验及效应因子的 ELISA 测定研究其抗肿瘤活性。结果 成功构建 IL15-NK92 细胞,具有较强的增殖活性,体外杀伤实验显示 CAR-IL15-NK92 细胞对 HER2 阳性肿瘤细胞的杀伤效应和效应因子分泌具有靶向特异性。结论 自分泌的 IL-15 可以促进 NK-92 细胞增殖,构建的靶向 HER2 的 CAR-IL15-NK92 细胞在体外显示出了良好的特异性杀伤能力,可以为后续开发以海洋鲨鱼纳米抗体为基础的 NK 细胞治疗药物奠定研究基础。 |
| English Summary: |
| Objective To explore the proliferative effect of autocrine interleukin-15 (IL-15) on NK-92 cells and the killing effect of chimeric antigen receptor (CAR) NK-92 cells targeting human epidermal growth factor receptor 2 (HER2) on related tumor cells. Methods The lentiviral vector was used to introduce IL-15 into NK-92 cells, the expression level of IL-15 was detected by ELISA, and its proliferative activity was detected by CCK-8 method. On this basis, the second-generation CAR targeting HER2 was introduced, the positive rate of CAR was detected by flow cytometry, and its anti-tumor activity was studied by in vitro killing assay and effecting factor ELISA. Results IL15-NK92 cells were successfully constructed with strong proliferative activity. In vitro killing experiments showed that CAR-IL15-NK92 cells had targeted specificity in killing effect and factor secretion of HER2-positive tumor cells. Conclusion Autocrine IL-15 can promote the proliferation of NK-92 cells, and the constructed HER2-targeting CAR-IL15-NK92 cells showed excellent specific killing ability in vitro, laying a foundation for the subsequent development of NK cell therapy drugs based on Marine shark nanoantibodies. |
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