College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, 30 Puzhu South Road, 211816, Nanjing, P. R. China.
Jiangsu Tripod Preclinical Research Laboratory Co. Ltd, 211816, Nanjing, China.
J Exp Clin Cancer Res. 2021 Nov 20;40(1):371. doi: 10.1186/s13046-021-02173-5.
Brain metastasis (BM) is thought to be related to the mortality and poor prognosis of non-small cell lung cancer (NSCLC). Despite promising development of NSCLC treatment, the treatment of NSCLC BM is still not optimistic due to the existence of the blood-brain barrier (BBB) that prevent drug penetration, as well as the short median survival time of the patients left for treatment. In this context, further development of quick and effective pre-clinical models is needed in NSCLC BM treatment. Here, we report a model system using zebrafish to promote the development of drugs for patients with NSCLC BM.
Three different NSCLC cell lines (H1975, A549 and H1299) were used to establish zebrafish BM models. The embryo age and cell number for injection were first optimized. Metastatic cells were observed in the brain blood vessels of zebrafish and were verified by hematoxylin-eosin (HE) staining. Then, the metastasis potentials of H1975 and A549 with manipulated microRNA-330-3p (miR-330-3p) expression were also investigated. Finally, sensitivities of H1975 and A549 to osimertinib and gefitinib were tested.
This zebrafish BM model could distinguish NSCLC cell lines with different BM potential. Over-expressed miR-330-p significantly improved the BM potential of the A549 cells while knockdown miR-330-p reduced the BM ability of the H1975 cells. Both osimertinib and gefitinib showed inhibition effect in zebrafish BM model with the inhibition rate higher than 50 %. H1975 cell showed much higher sensitivity to osimertinib rather than gefitinib both in vivo and in vitro.
We established zebrafish brain metastasis model for studying mechanism and treatment of NSCLC BM. This study provided a useful model for NSCLC brain metastasis that could be used to study the mechanism that drive NSCLC cells to the brain as well as identify potential therapeutic options.
脑转移（BM）被认为与非小细胞肺癌（NSCLC）的死亡率和预后不良有关。尽管 NSCLC 治疗取得了令人鼓舞的进展，但由于血脑屏障（BBB）的存在阻碍了药物渗透，以及留给治疗的患者中位生存时间较短，NSCLC BM 的治疗仍然不容乐观。在这种情况下，需要进一步开发快速有效的 NSCLC BM 治疗的临床前模型。在这里，我们报告了一种使用斑马鱼促进 NSCLC BM 患者药物开发的模型系统。
使用三种不同的 NSCLC 细胞系（H1975、A549 和 H1299）建立斑马鱼 BM 模型。首先优化胚胎年龄和注射细胞数。观察到转移细胞在斑马鱼的脑血管中，并通过苏木精-伊红（HE）染色验证。然后，还研究了表达受调控的 microRNA-330-3p（miR-330-3p）的 H1975 和 A549 的转移潜力。最后，测试了 H1975 和 A549 对奥希替尼和吉非替尼的敏感性。
该斑马鱼 BM 模型可区分具有不同 BM 潜力的 NSCLC 细胞系。过表达 miR-330-p 显著提高了 A549 细胞的 BM 潜力，而敲低 miR-330-p 降低了 H1975 细胞的 BM 能力。奥希替尼和吉非替尼在斑马鱼 BM 模型中均显示出抑制作用，抑制率均高于 50%。H1975 细胞在体内和体外对奥希替尼的敏感性均明显高于吉非替尼。
我们建立了用于研究 NSCLC BM 机制和治疗的斑马鱼脑转移模型。这项研究为 NSCLC 脑转移提供了一个有用的模型，可用于研究驱动 NSCLC 细胞进入大脑的机制以及确定潜在的治疗选择。
