Abstract
Objective
To investigate the mechanism by which Chinese medicine Shengmai Yin (SMY) reverses epithelial-mesenchymal transition (EMT) through lipocalin-2 (LCN2) in nasopharyngeal carcinoma (NPC) cells CNE-2R.
Methods
Morphological changes in EMT in CNE-2R cells were observed under a microscope, and the expressions of EMT markers were detected using quantitative real-time PCR (RT-qPCR) and Western blot assays. Through the Gene Expression Omnibus dataset and text mining, LCN2 was found to be highly related to radiation resistance and EMT in NPC. The expressions of LCN2 and EMT markers following SMY treatment (50 and 100 µ g/mL) were detected by RT-qPCR and Western blot assays in vitro. Cell proliferation, migration, and invasion abilities were measured using colony formation, wound healing, and transwell invasion assays, respectively. The inhibitory effect of SMY in vivo was determined by observing a zebrafish xenograft model with a fluorescent label.
Results
The CNE-2R cells showed EMT transition and high expression of LCN2, and the use of SMY (5, 10 and 20 µ g/mL) reduced the expression of LCN2 and reversed the EMT in the CNE-2R cells. Compared to that of the CNE-2R group, the proliferation, migration, and invasion abilities of SMY high-concentration group were weakened (P<0.05). Moreover, SMY mediated tumor growth and metastasis in a dose-dependent manner in a zebrafish xenograft model, which was consistent with the in vitro results.
Conclusions
SMY can reverse the EMT process of CNE-2R cells, which may be related to its inhibition of LCN2 expression. Therefore, LCN2 may be a potential diagnostic marker and therapeutic target in patients with NPC.
Keywords: epithelial-mesenchymal transition, lipocalin-2, nasopharyngeal carcinoma, radiation therapy, Shengmai Yin, Chinese medicine
Acknowledgement
The authors gratefully acknowledge generous support by of the Radiotherapy Department of Nanfang Hospital.
Footnotes
Supported by the National Natural Science Foundation of China (No. 82074132), Project of Administration of Traditional Chinese Medicine of Guangdong Province of China (No. 20213009) and the Science and Technology Plan Project of Guangzhou City (No. 202102080405)
Conflict of Interest
The authors declare no conflicts of interest.
Author Contributions
Wang ZT and Peng Y conducted experimental research, collected the test data, elucidated the results. Lou DD designed this research and wrote the manuscript. Zeng SY and Zhu YC took part in the experiment. Li AW and Lyu Y provided constructive suggestions. Fan Q and Zhu DQ directed the project and managed the funds. All authors read and approved the final version for publication.
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