The Study Of HDAC Regulation YAP/RON Signaling Pathway To Prevent Smoking-Induced Liver And Lung Metastases Of PDAC And Effects Of Emodin On It

Objective: 1.To observe cigarette smoke NNK and CSE in vivo and in vitro experiments effect on the growth of pancreatic cancer cell lines,to know their invasiveness,and the occurrence of liver and lung metastases.To find out that HDAC(Class I and II),especially HDAC4 inhibitors,prevent the EMT and invasion of smoking compounds.2.To study NNK and CSE effect on the protein level of HDAC4 and the level of Yes-associated protein 1(Yap)translocation to the nucleus,HDAC4 inhibitor significantly reversed this effect.3.To confirm NNK and CSE effect on the metastasis of pancreatic cancer cells to mice;and HDAC inhibitors block this effect.4.To observe the biological effects of emodin(EMO)on pancreatic cancer cell lines Bx Pc3 and Mia Pa Ca-2.And the effect of EMO on promoting EMT and invasion of smoking5.To investigate the effect of EMO on the expression of HDAC4,YAP and RON in cells and nucleus extraction.And the effect of EMO on smoking-induced liver and lung metastases of pancreatic cancer.Methods: Pancreatic cancer cells were treated with different doses of4-(methylnitro-s-amino)-1-(3-pyridyl)-1-butanone(NNK)and cigarette smoke extract(CSE)and cancer cell survival,markers of epithelial to mesenchymal transition(EMT)as well as invasion measured.In a syngeneic model of pancreatic cancer,mice were treated with NNK and progression of the disease including metastasis to liver and lung measured with and without inhibition of histone deacetylase(HDAC)of the classes I and II.Pancreatic cancer cells were treated with a certain dose of EMO.The morphological effects of EMO on pancreatic cancer cells were observed.Pancreatic cancer cells were treated with different doses of NNK and CSE.Cell viability,EMT and invasion weredetected.In a pancreatic cancer animal model,NNK,and EMO were added after one week of modeling,and to measure the metastasis to lung and liver.Results: Smoking compounds slightly but significantly increased cancer cell survival.They significantly promoted expression of EMT markers and promoted invasion.Inhibition of HDAC(class I and II),and especially HDAC4,prevented the pro-EMT and invasion effect of smoking compounds.NNK increased the protein level of HDAC4 and the level of Yes-associated protein 1(Yap)translocated to the nucleus.Inhibition of HDAC4 reversed this effect.Analysis of Yap binding proteins showed presence of HDAC4 in the Yap protein complex.NNK promoted metastasis of pancreatic cancer cells to mice lung;whereas,inhibition of HDAC prevented this effect.EMO could promote the morphological changes of pancreatic cancer cells and accelerate their apoptosis,inhibit smoking to some extent,increase the survival rate and invasiveness of pancreatic cancer cells and the effect of EMT;EMO could change the expression of HDAC4,Yap and RON signal proteins in pancreatic cancer cells and nucleus extraction.EMO can partially inhibit smoking-induced liver and lung metastasis of pancreatic cancer in animal model.Conclusion: We find that HDAC inhibittor can effectively reverse the pancreatic cancer cell line survival,invasion and EMT promoted by smoking.We showed for the first time in an animal model that smoking promotes metastasis of PDAC to the liver and lung.We found that HDAC4 is key mediator of this effect.Inhibiting HDAC4 is a promising strategy for preventing PDAC metastasis.EMO promotes apoptosis of pancreatic cancer cells,change the expression of HDAC/Yap/RON signal protein,and inhibits smoking in promoting liver and lung metastasis of pancreatic cancer.It provides a effective strategy for the clinical treatment of pancreatic cancer.