Based On The JNK And ERK Signaling Pathways,the Effect Of Juglone On The Apoptosis Of Lung Cancer A549 Cells Was Studied

Purpose:Based on the experimental study of JNK and ERK signaling pathways on the regulation of juglone on the apoptosis of A549 lung cancer cells,this study provides a new idea for Juglone as a treatment for non-small cell lung cancer,and provides a necessary theoretical basis for the clinical feasibility and research of Juglone.Method:In this study,A549 cells were cultured in vitro and the effects of Juglone at different concentrations and time were studied.Cell viability was detected by MTT assay,cell migration;Western blot assay was used to detect Caspase-3,Caspase-9,Bcl-2,Bax,Cyt C,pERK and pJNK protein levels were detected by scratch test and Transwell assay,apoptosis was detected by flow cytometry.In addition,we also used HE staining and immunohistochemical staining to investigate the effect of Juglone on nude mice transplanted tumor model.Results:(1)MTT assay showed that Juglone decreased A549 cell viability in a concentration-dependent manner.The optimal concentration was 20?mol/L.(2)Scratch test showed that the migration rate of A549 cells treated with20?mol/L Juglone for 24 h and 48 h was lower than that of the control group,and the migration of A549 cells was inhibited significantly at 48 h.(3)Transwell results showed: With the increase of the concentration of Juglone,the migration quantity of A549 cells decreased with the increase of the concentration of Juglone detected by Transwell for 48 h.(4)The results of Flow Cytometry showed that Juglone inhibited the apoptosis of A549 cells at 48 h,and increased with the drug concentration.(5)Western blot showed that Juglone inhibited the expression of Bcl-2and ERK proteins.The expression levels of Bax,Casapse-3,Casapse-9,Cyt C and pJNK were significantly increased.(6)HE staining was used to observe the section of xenograft tumor in nude mice.Under the 200-fold field of view,the tumor cells in the control group were of different sizes,closely arranged,vigorous growth,and intact cell morphology,showing the typical morphology of tumor cells.Compared with the control group,the tumor tissues of the Juglone group showed necrotic areas,unconspicuous nucleus blue-staining,fragmented aggregation of cells,and unclear tumor cell growth boundaries.(7)Immunohistochemical staining showed that the expression of Bcl-2in the Juglone group was lower than that in the control group.The expression of Bax,Casapse-3 and Casapse-9 in the Juglone group was higher than that in the control group.Conclusion:By inhibiting the expression of bcl-2 and pERK proteins,the increased expressions of Bax,Casapse 3,Casapse 9,Cyt C and pJNK promoted the apoptosis of A549 cells.The injection of Juglone can inhibit the growth of A549 transplanted tumor,and inhibit the growth of tumor by inhibiting bcl-2and up-regulating Bax,Casapse 3 and Casapse 9.Meanwhile,Juglone inhibited the proliferation and migration of A549 cells to inhibit tumor growth.It can be concluded that Juglone has the effect of inhibiting tumor growth.