| Objective: With the increased incidence of cancer,as one of the most common malignancies in the digestive system tumors,pancreatic cancer(pancreatic,cancer,PC)currently has a very high morbidity and mortality.It is the eighth highest incidence of tumors,and ranked fourth in the United States cancer mortality.Previous studies have statistics show that the incidence of pancreatic cancer in China has increased year by year:seventh of the incidence and mortality of malignant tumors in China is the sixth.Both at home and abroad for the future of biological therapy is constantly open,so to clarify the pathogenesis of pancreatic cancer,to find new gene therapy targets become particularly important.We have found out the mechanism of the HIPPO pathway in gastric cancer and cancinomor of colon.there have been numerous studies to investigate the biological characteristics of pancreatic cancer in various pathways of the regulation.To further clarify the regulatory mechanism of various factors in the HIPPO signaling pathway in the occurrence and development of pancreatic cancer.To investigate the role and metastasis of YAP、TEAD、LATS1、MST1and Cyclin E in HIPPO-YAP pathway in pancreatic tissue,pancreatic cancer tissue and adjacent pancreatic tissue.The relationship between micro RNA and tumor has become a hot spot.As for the relationship between micro RNA186 and tumor,it is found that it can regulate the occurrence and development of tumor.The present study is to determine the expression of micro RNA(miRNA or miR)-186 in tumor tissues and peripheral blood of patients with pancreatic cancer(PC),as well as its mechanism of regulation.Method: 1.Collects 31 patients with pancreatic cancer underwent surgical resection from 2012 to 2014 in the Hepatobiliary surgery of the Affiliated Hospital of Qingdao Universit and Qingdao municipal hospital,All the cases were confirmed as primary pancreatic ductal adenocarcinoma,No chemotherapy or radiotherapy before operation.There were 17 males and 41 females,aged 36-79 years,with an average age of 57 ±0.5years old.21 cases were in the part of pancreatic head,and the other 10 cases were in the part of pancreatic body and tail.Histological type: There were 17 cases of poorly differentiated adenocarcinoma,8 cases were moderately differentiated,and 6 cases were well differentiated adenocarcinoma.At the same time,31 cases of adjacent cancer tissues were taken from the tumor adjacent.12 cases of normal pancreatic tissue due to trauma.There were 7 males and 5 females,aged 43-76 years,with an average age of 59 +0.5years old.Using the methods of RT-PCR and Western Blot todetect the expression of each factor in HIPPO-YAP pathway of thirty-one pancreatic cancer tissues and adjacent pancreatic tissues,tewlve normal pancreatic tissues.2.A total of 46 patients with PC who underwent surgery between 2012 and 2015 were included in the present study in the Hepatobiliary surgery of the Affiliated Hospital of Qingdao Universit and Qingdao municipal hospital.In addition,46 healthy subjects were included as controls.PC group included 26 males and 20 females aged between 40 and 76 years(median age,61 years).Control group included 26 males and 20 females aged between 40 and 75 years(median age,58 years).There were 22 cases of poorly differentiated adenocarcinoma,14 cases were moderately differentiated,and 10 cases were well differentiated adenocarcinoma.tumor with node metastasis: period1 were 22caases;period2 were 20 cases;period3 were 4 cases.Lymph node metastasis in 26 cases.All PC patients were having onset of PC for the first time and had no history of uses of hormones,Chinese medicine,or chemoradiotherapy.Quantitative real-time polymerase chain reaction was used to measure the expression of mRNA and miRNA.Western blotting and enzyme-linked immunosorbent assay were used to determine protein expression.Bioinformatics was employed for the prediction of target gene of miR-186,while dual luciferase reporter assay was carried out to identify direct binding of miR-186 with YAP1 mRNA.Human pulmonary aortic endothelial cells(HPACs)were transfected with ago-miR-186.YAP1 expression in HPACs was silenced by si RNA.MTT assay was used to evaluate the proliferation of HPACs.Result: The expressions of MST1 and LATS2 mRNA in pancreatic carcinoma was lower than in normal pancreatic tissue(P < 0.001),on the contrary,The expressions of YAP,TAZ and TEAD1 in pancreatic cancer is higher than that in normal pancreatic tissue(P <0.001).The expression level of MAT1 and LATS2 mRNA in pancreatic cancer tissue to paracancerous tissue were increased,while the expression level of YAP,TAZ and TEAD1 from pancreatic cancer to adjacent tissues were decreased.In addition,Statistical analysis showed that the expression of Hippo signaling pathway was related to tumor differentiation,lymph node metastasis and TNM stage.YAP1 mRNA and protein expression was elevated in PC.In addition,expression of miR-186 in PC was down-regulated.miR-186 regulated the expression of YAP1 by binding with the 3’-UTR of YAP1.Elevated expression of miR-186 inhibited the proliferation of HPACs by down-regulating the expression of YAP1.Decreased expression of YAP1 by si RNA inhibited the proliferation of HPACs.Conclusion: The expression of MST1 and LATS1 was decreased in pancreatic cancer tissues,While the expression of YAP,Cyclin E and TEAD1 was increased in pancreatic cancer tissues,and the HIPPO-YAP pathway was related to the development of pancreatic cancer.The present study demonstrates that YAP1 is up-regulated in tumor tissues and blood of PC patients,and this may be related with the down-regulation of miR-186.In addition,miR-186 may affect the occurrence and development of PC by controlling the proliferation of PC cells via YAP1. |
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