Effect of knocking down exosomes derived from Nei endonuclease Ⅷ-like 3 on migration,autophagy and temozolomide resistance in glioma cells
Objective To investigate the expression of Nei endonuclease Ⅷ-like 3(NEIL3)in glioma and its effects on the growth,migration,autophagy,and Temozolomide resistance of glioma cells,as well as to elucidate its regulatory mechanisms.Methods A total of 15 samples of human glioma tissue and adjacent tissue were collected from the Neurosurgery Department of the First Affiliated Hospital of Zhengzhou University.This study assessed the expression of NEIL3 in glioma tissues via qPCR assays,and utilized gene knockdown techniques to reduce NEIL3 expression in glioma cells and extract and purify NEIL3-derived exosomes.Cell proliferation was assessed using the cell counting kit-8(CCK-8)assay and colony formation assay,and cell migration was evaluated using the Transwell assay.Autophagy-related pro-tein expression levels were detected using immunoblotting and immunofluorescence techniques.The sensi-tivity of glioma cells to Temozolomide was determined by flow cytometry,and the activity of the phosphati-dylinositol 3 kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway was analyzed by immunoblotting.T-test was used to compare the differences between two groups,and one-way analysis of variance(ANOVA)was used for multi group comparisons.Results The RNA ex-pression level of NEIL3 in glioma tissues(1.00±0.04)was significantly higher than that in adjacent nor-mal tissues(0.47±0.11,P＜0.01).Knockdown of NEIL3-derived exosomes could inhibit the growth[(55.00±0.06)％vs.(100.00±0.53)％,P＜0.01]and colony formation(49.67±11.55 vs.197.30±44.24,P＜0.01)of glioma cells,suppress autophagy in glioma cells,improve the resistance of gli-oma cells to temozolomide,and inhibit the PI3K/Akt/mTOR signaling pathway in glioma cells.Conclusion Knockdown of NEIL3-derived exosomes can inhibit glioma cell migration and autophagy and improve Temozo-lomide sensitivity.
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