| Objective:To study the expression of ISYNA1 and association of ISYNA1 with clinicopathological significance in pancreatic ductal adenocarcinoma(PDAC).Methods:Collecting clinical data and specimens of PDAC patients in Department of General Surgery,the First Hospital of China Medical University from March 2008 to December 2017;39 patients who had incomplete clinical or follow-up data and died in perioperative period were excluded,and 68 patients were finally analyzed,39 males and29 females,33-81(median 59)years old.The expression of ISYNA1 in 68 paraffin embedded PDAC specimens was detected by immunohistochemistry(IHC),in which 34had paired non—cancerous pancreatic tissues,the difference of ISYNA1 expression in PDAC and normal pancreatic tissues were compared,and the relationship between ISYNA1 expression and clinicopathological parameters was analyzed;and we estimated the correlation between ISYNA1 and p53 in 48 PDAC specimens.At the same time,another 17 paired fresh PDAC specimens were collected between September 2016and August 2017,qRT-PCR and Western Blot were used to examine the expression of ISYNA1 mRNA and protein level in 17 paired fresh PDAC specimens and adjacent non—cancerous pancreatic tissues,respectively;and the expression difference of ISYNA1 in paired PDAC and normal pancreatic tissues were further identified.siRNA interference was used to knockdown the expression of p53 in Capan-2,SW1990 and Miapaca-2 cells,and association of p53 with ISYNA1 expression was explored.Statistical methods included Student's test,?~2test,Kaplan-Meier curve,Log-Rank test and Pearson analysis.Result:IHC showed ISYNA1 mainly expressed in cytoplasm;the high expression rate of ISYNA1 in 34 normal pancreatic tissues was 55.9%(19/34),however the high expression rate of ISYNA1 in 68 PDAC tissues was 38.2%(26/68),and the expression of ISYNA1in normal pancreatic tissues(6.012±3.428)was significantly higher than that in PDAC tissues(3.681±2.198)(t=3.611,P=0.001).In 17 paired fresh PDAC specimens,ISYNA1 mRNA expression in non-cancerous pancreatic tissues(3.721±2.234)was obviously higher than that in PDAC tissues(5.889±1.607)(t=-4.636,P<0.01),and ISYNA1 protein level in non-cancerous pancreatic tissues(0.815±0.418)was similarly higher than that in PDAC tissues(0.517±0.240)(t=2.948,P=0.009).?~2 test showed the expression of ISYNA1 was negatively associated with tumor invasion depth(?~2=7.534,P=0.030)and vascular invasion(?~2=5.048,P=0.043);but had no relationship with age,gender,tumor location tumor size,differentiation,lymph node metastasis,TNM stage,postoperative liver metastasis and perineural permeation.In 68 PDAC patients,postoperative pathology of 48 patients tested the expression level of p53(mutant p53).Comprehensive analysis of ISYNA1 and p53 expression in the 48 PDAC showed 8patients expressed both high level of ISYNA1 and p53;11 patients synchronously expressed low level of ISYNA1 and p53;20 patients expressed high level of p53 but low ISYNA1;9 patients expressed low level of p53 but high ISYNA1.Pearson analysis indicated that there was no relationship between ISYNA1 and mutant p53(?2=1.377,P=0.359).In p53 wild-type Capan-2 and SW1990 cells,Knockdown of p53 significantly down regulated ISYNA1 expression,whereas had no effect on ISYNA1 expression in p53 mutant Miapaca-2 cells.Kaplan-Meier survival analysis and Log-Rank test indicated patients with negative ISYNA1 expression had a shorter median survival time and poorer prognosis(?~2=4.953,P=0.026).Conclusion:The expression of ISYNA1 was associated with clinicopathological parameters of PDAC patients,and its expression in PDAC tissues is significantly decreased,which is associated with the prognosis of PDAC patients.The expression of ISYNA1 is only related to wild type p53,and has no relationship with mutant p53.Abnormal expression of ISYNA1 may play an important role in the progression of PDAC. |
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