Effects Of Recombinant Newcastle Disease Virus (rL-hIFN-?1) On Endoplasmic Reticulum Stress,Autophagy And Apoptosis In Cell Line Of Small Cell Lung Cancer

Objective:To investigate the effect of?recombinant newcastle disease virus expressing IL-29?rL-hIFN-?1 on endoplasmic reticulum stress,autophagy and apoptosis in the small cell lung cancer?SCLC?cell line H446 cells.Methods:SCLC H446 cells were randomly divided into control group,newcastle disease virus NDV group and rL-hIFN-?1 group.The latter two groups were transfected with NDV and rL-hIFN-?1 respectively.The cell viability was detected by CCK-8 assay;Cell supernatant was detected by ELISA,the content of IL-29 was detected.Western blot and immunofluorescence were used to detect the expression of endoplasmic reticulum stress-related protein?GRP78,CHOP?.After pre-treated with 4-PBA?endoplasmic reticulum stress inhibitor?,3-MA?autophagy inhibitor?,rapamycin?autophagy enhancer?and siRNA knock-out of Bcl-2,western blot was used to detect the expression of endoplasmic reticulum stress-related proteins?GRP78,CHOP?,autophagy-related proteins?LC3 I/II?and cleaved-caspase-3 expression both in treated and control groups.Results:1.After r L-hIFN-?1 and NDV infection of H446 cells,CCK-8 results showed that with the increase of virus dilution,cell activity increased,which rL-hIFN-?1 group in 10^-5,10^-6 cell viability>80%.After determining the dilution concentration of the virus,the virus titer was measured.2.Western blot and ELISA results showed that H446 cells stably expressed IL-29,indicating successful virus infection.3.Immunofluorescence and Western blot results showed that the expression of endoplasmic reticulum stress-related protein?GRP78,CHOP?was significantly increased after rL-hIFN-?1 and NDV infection?P<0.05?,and rL-hIFN-?1 is more effective than NDV.4.Western blot showed that endoplasmic reticulum stress?GRP78,CHOP?,autophagy?LC3 I/II?and apoptosis?cleaved-caspase3?proteins expression were significantly higher than that of the control group after treatment with 4-PBA and 3-MA?P<0.05?.5.After treatment with siRNA and Rapa,the expression of endoplasmic reticulum stress?GRP78,CHOP?,autophagy?LC3 I/II?and apoptosis?cleaved-caspase3?proteins were higher than that of the control group?P<0.05?.Conclusion:1.rL-hIFN-?1 can induce endoplasmic reticulum stress,autophagy and apoptosis in SCLC H446 cells.2.4-PBA and 3-MA can inhibit endoplasmic reticulum stress,autophagy and apoptosis induced by r L-hIFN-?1 in H446 cells.3.After treated with siRNA and Rapa,it can enhance the endoplasmic reticulum stress,autophagy and apoptosis induced by rL-hIFN-?1 in H446 cells.