Experimental study on Derlin-1 promoting paclitaxel resistance in hypopharyngeal cancer by regulating the PI3K/Akt signaling pathway and autophagy
DENG Yaping
TANFengwu
LI Kehua
Abstract：Objective To investigate the role of degradation in endoplasmic reticulum proteim 1(Derlin-1),a key protein of endoplasmic reticulum-associated degradation(ERAD),in the formation of paclitaxel resistance in hypopharyngeal carcinoma and its regulatory mechanism on the PI3K/Akt signaling pathway and autophagy.Methods The drug-resistant hypopharyngeal carcinoma Fadu cell line was constructed by paclitaxel gradient induction,and the IC50 and resistance index(RI)were determined by CCK-8.The experiment was divided into the control group,the paclitaxel group(80 nmol/L),the Derlin-1 silencing group(siRNA interference),and the Derlin-1 silencing+paclitaxel group.Cell viability was detected by the CCK-8 method,and the apoptosis rate was detected by flow cytometry.Western blotting was used to detect the protein expressions of Derlin-1,Bax,Bcl-2,p-PI3K/PI3K,p-Akt/Akt,LC3-Ⅱ/Ⅰ and Beclin-1.Observation of autophagosome and autophagolysosome formation by mCherry-GFP-LC3B dual-fluorescence adenovirus combined with confocal microscopy.Results The IC50 of drug-resistant cells increased from 8.438 nmol/L in parental cells to 73.330 nmol/L,the RI was 8.69,and the Derlin-1 protein was significantly upregulated.The cell viability in the paclitaxel group decreased to approximately 65%,and the apoptosis rate was about 18.6%.Cell viability in the Derlin-1 silencing+paclitaxel group further decreased to approximately 42%,and the apoptosis rate increased to approximately 36.2%.Compared with the control group,in the paclitaxel group,Bax increased,Bcl-2 decreased,p-Akt/Akt and p-PI3K/PI3K decreased,and LC3-Ⅱ/Ⅰ and Beclin-1 increased.In the Derlin-1 silencing+paclitaxel group,these differences were all greater than those in the paclitaxel group,manifested as a further increase in Bax,a further decrease in Bcl-2,a lower phosphorylation level of the PI3K/Akt signaling pathway,a higher expression of autophagy-related proteins,and a significant increase in the number of autophagosomes and autophagolysosomes.Conclusion Derlin-1 may promote paclitaxel resistance in hypopharyngeal cancer by activating the PI3K/Akt signaling pathway and regulating the autophagy process.Silencing the expression of Derlin-1 can reverse the drug resistance phenotype and enhance paclitaxel-induced apoptosis and autophagy,suggesting that Derlin-1 may serve as a potential therapeutic target to overcome drug resistance.
Keywords：Hypopharyngeal carcinomaDegradation in endoplasmic reticulum proteim 1(Derlin-1)PaclitaxelPI3K/Akt signaling pathwayAutophagy
Publication Date：2025-09-28
Online Publishing Date：2025-10-28(First online date of this platform, not the publication date of the document)
Pages：6( 833-838 )