Study On The Relationship Between PD-L1 And Autophagy And Proliferation Of Lung Cancer Cells Induced By Hypoxia

Objective: In recent years,the treatment of advanced lung cancer has changed,gradually giving priority to immunotherapy and other treatments as auxiliary collaborative therapy.the local application of tumor immunotherapy drugs,especially the anti-PD-L1 and its receptor drugs show obvious anti-tumor effects so that cancer patients continue to benefit.Early clinical exploration found that anti-PD-L1 and its receptor drugs have clear anti-tumor activity,good immune induction response,and so on.However,with the deepening of treatment,immune-related adverse reactions have become increasingly prominent,some of which are even fatal.We know that a tumor has its complex and unique tumor microenvironment,and all therapeutic measures generally need to act on the tumor microenvironment before it can have an anti-tumor effect.Previous studies have found that hypoxia,as one of the most important features of the tumor microenvironment,generally exists in solid tumor tissues.However,the mechanism of the correlation between the anoxic microenvironment and the immune checkpoint is still unclear,and further research is crucially required.In this research,we analyzed the effects of hypoxia minienvironment on autophagy and expansion of lung carcinoma cells and observed the changes of molecular expression of autophagyrelated pathway proteins and hypoxia-inducible factor in lung carcinoma cells under different hypoxia levels,as well as their effects on cell activity,to explore the character of PD-L1 in the hypoxia-induced autophagyrelated pathway.It provides a new treatment idea for clinical front-line researchers of tumor microenvironment and immunotherapy.At the same time,we can try to combine PD-L1 immunosuppressants,an autophagy inhibitor,and hypoxia therapy to explore the synergistic effect and provide a new treatment for immunotherapy of lung cancer in the future.Methods: Through the hypoxia culture environment,taking the protein expression level of hypoxia-inducible factor as the reference standard,a hypoxia microenvironment model of lung cancer was established.On this basis,the control band and the experimental band were set up,the control band was the Normoxia,the experimental band was divided into hypoxia band and hypoxia + 3-MA drug intervention band(Hypoxia and Hypoxia+3-MA,respectively),in which the control band was normoxic culture environment,the experimental band was hypoxia culture environment,hypoxia + 3-methyladenine band was treated with 3-MA for 12 hours,24 hours and 48 hours cooperatively.The proliferation action of lung carcinoma cells was measured by CCK-8 at different time points,the definition of autophagy and hypoxia-related proteins under different conditions were recognized by WB,and the m RNA definition levels of AKT,LC3-Ⅱ and PD-L1 were studied by real-time PCR.Finally,the laboratory data were gathered and related analytical investigation was carried out.Results: By changing the hypoxia condition,we constructed a stable hypoxia model of lung cancer.In the hypoxia band,with the augmentation of hypoxia time(12h-48h),compared with the normoxic band,the protein declaration of HIF-1 α and LC3-II enlarged continuously,and the gene level of LC3-Ⅱ,PD-L1,and AKT enlarged gradually.At the same time,the proliferative activity of H1299 lung cancer cells also increased gradually.After drug intervention with 3-MA,a specific autophagy inhibitor,the molecular expression levels of HIF-1 α,LC3-Ⅱ,PD-L1,and AKT in the hypoxia + 3-MA band were accordingly lesser than those in the hypoxia band,and the activity of cell proliferation was accordingly degenerated.Conclusion: Hypoxia tumor microenvironment will promote H1299 lung cancer cells to undergo a series of adaptive changes.There was a significant correlation between autophagy pathway protein and PDL1 in H1299 lung cancer cells induced by hypoxia.PD-L1 may be involved in the expression of the autophagy-related pathway in H1299 lung cancer cells,which may be related to the PI3K/AKT pathway.3-methyladenine can accordingly inhibit the expression of autophagy pathway protein and PDL1 in lung carcinoma cells,and also suppress the reproduction of lung cancer cells.Hypoxia may promote the reproduction of H1299 lung carcinoma cells by stimulating PI3K/AKT autophagy roadway and upregulating the expression of immune checkpoint PD-L1 and autophagy roadway protein LC3-II.