| Objective:To investigate the effect of（recombinant newcastle disease virus expressing IL-29）rL-hIFN-λ1 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-λ1 group.The latter two groups were transfected with NDV and rL-hIFN-λ1 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-λ1 and NDV infection of H446 cells,CCK-8 results showed that with the increase of virus dilution,cell activity increased,which rL-hIFN-λ1 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-λ1 and NDV infection（P<0.05）,and rL-hIFN-λ1 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-λ1 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-λ1 in H446 cells.3.After treated with siRNA and Rapa,it can enhance the endoplasmic reticulum stress,autophagy and apoptosis induced by rL-hIFN-λ1 in H446 cells. |
