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 68 PDAC patients at Department of General Surgery, the First Hospital of China Medical University from March 2008 to December 2017.There were 39 males and 29 females, aged 33 to 81 years(median 59 years).The expression of ISYNA1 in 68 paraffin embedded PDAC specimens was detected by immunohistochemistry,in which 34 had paired non-cancerous pancreatic tissues,the relationship between ISYNA1 expression and clinicopathological parameters was analyzed; and the correlation between ISYNA1 and p53 in 48 PDAC specimens were estimated.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.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,χ(2) test, Kaplan-Meier curve, Log-rank test and Pearson analysis, respectively. Results: Immunohistochemistry results showed that the expression of ISYNA1 in PDAC(3.681±2.198)was significantly lower than that in normal pancreatic tissues(6.012±3.428)(t=-3.611,P=0.001).In 17 paired fresh PDAC specimens,ISYNA1 mRNA expression in non-cancerous pancreatic tissues(()ΔC(T): (3.721±2.234)was obviously higher than that in PDAC tissues ()ΔC(T): (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).χ(2) test showed the expression of ISYNA1 was negatively associated with tumor invasion depth(χ(2)=7.534,P=0.030)and vascular invasion(χ(2)=5.048,P=0.043);Pearson analysis showed there was no relationship between ISYNA1 and mutant p53(χ(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(χ(2)=4.953, P=0.026). Conclusions: The expression of ISYNA1 in PDAC tissues is significantly decreased,which is associated with the prognosis of PDAC patients,it 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.
目的： 探讨ISYNA1在胰腺导管腺癌（PDAC）中的表达及其与临床病理学参数的关系。 方法： 收集2008年3月至2017年12月中国医科大学附属第一医院普通外科收治的68例PDAC患者的临床资料和术后标本。男性39例，女性29例，年龄33~81岁（中位数59岁）。运用免疫组化方法检测68例PDAC石蜡标本中ISYNA1的表达水平，其中34例有配对的癌旁组织，统计其与临床病理学参数的相关性，并分析48例PDAC患者中ISYNA1与p53表达的相关性；再应用实时定量PCR（qRT-PCR）和免疫印迹法分别检测17例配对冷冻保存的新鲜PDAC标本中ISYNA1 mRNA和蛋白的表达水平。利用siRNA干扰技术抑制三种胰腺癌细胞株Capan-2、SW1990和Miapaca-2中p53基因的表达，探讨p53与ISYNA1表达的关系。分别采用t检验、χ(2)检验、Kaplan-Meier法、Log-kank检验和Pearson相关系数分析等方法进行统计学分析。 结果： 免疫组化检测结果显示，ISYNA1在癌组织的表达水平（3.681±2.198）明显低于癌旁组织（6.012±3.428）（t=-3.611，P=0.001）。在17例配对的新鲜组织标本中，癌旁组织的ISYNA1 mRNA表达水平（ΔCT值为3.721±2.234）高于癌组织（ΔCT值为5.889±1.607）（t=-4.636，P<0.01）；癌旁组织的ISYNA1蛋白表达水平（0.815±0.418）亦高于癌组织（0.517±0.240）（t=2.948，P=0.009）。相关性分析结果显示，ISYNA1的表达水平与肿瘤浸润深度（χ(2)=7.534，P=0.030）和血管浸润（χ(2)=5.048，P=0.043）相关，而与突变型p53表达无关（χ(2)=1.377，P=0.359）；在p53野生型Capan-2和SW1990细胞中，敲除p53后ISYNA1表达明显降低，而在p53突变型Miapaca-2细胞中，敲除p53后ISYNA1表达未受影响。Kaplan-Meier生存分析和Log-rank检验结果显示，ISYNA1低表达的患者中位生存时间较短（χ(2)=4.953，P=0.026），预后较差。 结论： ISYNA1在PDAC中呈明显低表达，有助于评估患者的预后，且其表达仅与野生型p53有关，与突变型p53无关；ISYNA1的表达异常可能与PDAC的恶性进程密切相关。.
Keywords: Biological markers; Pancreatic neoplasms; Prognosis.