Study On The Mechanism Of Qi Lian Fu Zheng Capsule Inhibiting The Proliferation And Metastasis Of Lung Cancer Based On NF-κB-related Inflammatory Signal Pathway

Objective: To study the mechanism of Qi lian fu zheng Capsule regulating the NF-κB inflammatory signal pathway to inhibit the proliferation and metastasis of lung cancer in tumor inflammatory microenvironment,and to explore the construction of a“cancer-toxin”-centered lung cancer diagnosis and treatment ideas and system.Methods: According to the basis of previous research,the drug-containing serum solution of Qi lian fu zheng capsule was prepared.Human lung cancer A549 cells were cultured in vitro,and the co-culture system of A549 cells and macrophages was established.To observe the effect of inflammatory environment on the morphology of A549 cells and the intervention effect of drug-containing serum of Qi lian fu zheng capsule on the migration ability of A549 cells under co-culture conditions.The effect of drug-containing serum of Qi lian Fu zheng capsule on the invasion ability of A549 cells was observed by Transwell chamber invasion test.The effect of drug-containing serum of Qi lian fu zheng capsule on the concentration of TNF-α and IL-6 in co-culture supernatant was detected by ELISA method.The effect of drug-containing serum of Qi lian fu zheng capsule on the expression of NF-κB pathway related genes and EMT related genes mRNA expression in A549 cells was observed by Real-Time PCR method.Western blot was used to observe the effect of drug-containing serum of Qi lian fu zheng capsule on the expression of NF-κB pathway related protein and EMT related protein in A549 cells under co-culture conditions.Results: Under the condition of inflammatory co-culture,lung cancer A549 cells were fibroid,which returned to epithelioid after adding the drug-containing serum of Qi lian fu zheng capsule.MTT test,scratch test and Transwell invasion test showed that the drug-containing serum of Qi lian fu zheng capsule could inhibit the proliferation,migration and invasion of A549 cells under co-culture conditions.While the drug-containing serum of Qi lian fu zheng capsule could down-regulate the concentration of TNF-α and IL-6 in the supernatant of co-culture.Real-Time PCR experiment showed that the drug-containing serum of Qi lian fu zheng capsule could down-regulate the expression of CHUK,IKBKB,CDH2,Rel A,VIM and SANIL1 mRNA,and up-regulate the expression of CDH1 and SNAIL2 in A549 cells under co-culture conditions,suggesting that the drug-containing serum of Qi lian fu zheng capsule could interfere with the expression of NF-κB and EMT protein related genes.Western blot experiment showed that the drug-containing serum of Qi lian fu zheng capsule could down-regulate the protein expression of NF-κB,IKKα,N-cad,VIM and Snail,and up-regulate the protein expression of IKKβ and E-cad in A549 cells under co-culture conditions,suggesting that the occurrence of EMT may be inhibited through NF-κB pathway,and then inhibit the invasion and metastasis of lung cancer cells.Conclusion: Qi lian fu zheng capsule can inhibit the proliferation,migration,invasion and metastasis of A549 cells under co-culture conditions.The mechanism may be related to down-regulating the level of inflammatory cytokines,thereby inhibiting the activation of NF-κB signal pathway and the formation of epithelial-mesenchymal transformation.