Study On The Mechanism Of Buzhong Yiqi Recipe On Laryngeal Squamous Cell Carcinoma Based On AKT1\NF-κB Signaling Pathway

Objective: To explore the effect of Buzhong Yiqi recipe on the proliferation of human laryngeal squamous cell carcinoma(LSCC)and the expression of AKT1 and NF-κB,and to elucidate the mechanism of Buzhong Yiqi recipe in inhibiting the proliferation and invasion of LSCC cells based on AKT1 / NF-κB signaling pathway.Method:(1)preparation of experimental serum: Twenty male Sprague Dawley rats,weighing 300±10g,were randomly divided into two groups: blank serum control group and Buzhong Yiqi recipe experimental serum group,with10 rats in each group,once a day,with a dose of 10 ml / kg each time.The blank serum control group was given normal saline by gavage,and the Buzhong Yiqi recipe experimental serum group was given classical formula Buzhong Yiqi recipe by gavage.After continuous gavage for 5 days,blood was taken from abdominal aorta and stored in-80℃ refrigerator.(2)Study on the proliferation of human laryngeal squamous cell carcinoma(Fa Du,SCC-4)by serum containing Buzhong Yiqi recipe,The experiment was divided into five groups:(1)control group: 100% ordinary medium;(2)blank serum group: 90%ordinary medium + 10% blank serum;(3)5% medicated serum group: 95%ordinary medium + 5% medicated serum;(4)10% medicated serum group:90% ordinary medium + 10% medicated serum;(5)20% medicated serum group: 80% ordinary medium+20% medicated serum.Different concentrations of Buzhong Yiqi recipe containing serum and blank serum were added to collect the supernatant and cells.CCK8 method was used to detect the proliferation of human laryngeal squamous cell carcinoma(Fa Du,SCC-4);(3)study on the mechanism of Buzhong Yiqi recipe containing serum on human laryngeal squamous cell carcinoma: Western blot method was used to detect the protein expression of AKT1,p-akt1,NF-κB.Spss22.0 software was used to analyze the data.Result:(1)Buzhong Yiqi recipe could delay the proliferation of human laryngeal squamous cell carcinoma(Fa Du,SCC-4)in all concentration groups,and the cell activity decreased to the lowest after 72 hours of intervention.The best concentration of Buzhong Yiqi recipe was 20%.There were significant differences in the cell proliferation activity rate between the drug containing serum groups and the control group(P < 0.05).(2)Compared with the control group,the expression of NF-κ B protein in Buzhong Yiqi recipe containing serum groups decreased,with significant difference(P < 0.05);the expression of NF-κ B protein in 5% Buzhong Yiqi recipe containing serum group decreased significantly,followed by 20% Buzhong Yiqi recipe containing serum group,with significant difference between the experimental groups(P <0.05).(3)Effect of serum containing different Buzhong Yiqi recipe on AKT1 signal pathway in human laryngeal squamous cell carcinoma: the expression of p-akt1 and AKT1 protein in 20% serum containing Buzhong Yiqi recipe group was significantly lower than that in the control group(P < 0.05).Conclusion:Buzhong Yiqi recipe containing serum can inhibit the proliferation of human laryngeal squamous cell carcinoma(Fa Du,SCC-4).NF-κB is the key regulatory target of its occurrence and development.Its mechanism may be through down regulating the expression of NF-κB protein to inhibit the proliferation of laryngeal squamous cell carcinoma.