Abstract
It has been reported that chemokine CX3CL1 can regulate various tumours by binding to its unique receptor CX3CR1. However, the effect of CX3CL1‐CX3CR1 on the lung adenocarcinoma and lung squamous cell carcinoma is still unclear. Here, we showed that CX3CL1 can further invasion and migration of lung adenocarcinoma A549 and lung squamous cell carcinoma H520. In addition, Western blot and immunofluorescence test indicated CX3CL1 up‐regulated the phosphorylation level of cortactin, which is a marker of cell pseudopodium. Meanwhile, the phosphorylation levels of c‐Src and c‐Abl, which are closely related to the regulation of cortactin phosphorylation, are elevated. Nevertheless, the src/abl inhibitor bosutinib and mutations of cortactin phosphorylation site could inhibit the promotion effect of CX3CL1 on invasion and migration of A549 and H520. Moreover, these results of MTT, Hoechst staining and Western blot suggested that CX3CL1 had no effect on the proliferation and apoptosis of A549 and H520 in vitro. The effects of CX3CL1 were also verified by the subcutaneous tumour formation in nude mice, which showed that it could promote proliferation and invasion of A549 in vivo. In summary, our results indicated that CX3CL1 furthered invasion and migration in lung cancer cells partly via activating cortactin, and CX3CL1 may be a potential molecule in regulating the migration and invasion of lung cancer.
Highlights
It is consistent with the results that protein level of PCNA showed no significant difference in each group (Figure 1E-H). These results suggested that 50 ng/mL or 100 ng/mL CX3CL1 has no obvious effect on the proliferation of lung cancer cells
We focused on exploring the effect of soluble CX3CL1 in regulating lung cancer A549 and H520 cells
Based on the preliminary understanding of the role of CX3CL1-CX3CR1 on lung cancer cells obtained in this study, we will further explore the effect of CX3CL1CX3CR1 on the development of lung cancer from the aspect of tumour microenvironment
Summary
Forty data of lung cancer patients were analysed. Between 2018 and 2019, the Pathology Archive of Chongqing Medical University First Affiliated Hospital (Chongqing, China) provided 40 lung cancer sections with approval of Chongqing Medical University Ethics Committee, and these 40 lung cancer specimens were used to detected p-cortactin 421 expression level by immunohistochemistry. To demonstrate the effect of CX3CL1 on lung cancer in vivo, A549 cells with the number of 5 × 106 together with 500 ng CX3CL1 were implanted into the nude mice to construct the xenograft model It is observed in the in vivo experiments that the tumour formation and growth was increased by adding with CX3CL1 compared to control mice (Figure 4A-C), and immunohistochemistry results showed that p-cortactin 421 and MMP-3 were up-regulated in CX3CL1 group (Figure 4D). The results of transwell with matrigel showed that the number of transmembrane cell in the bosutinib group was dramatically reduced compared to control (Figure 7F,G)
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