摘要
目的 观察酪氨酸蛋白磷酸酶非受体型3（PTPN3）在结直肠癌（CRC）中的表达,探讨其表达下调的机制.方法 用免疫组织化学方法检测112例CRC组织及其配对组织中的表达,荧光素酶实验分析微小RNA-95（miR-95）对PTPN3的调控作用,定量反转录.聚合酶链反应（RT-PCR）技术及Western blot技术检测PTPN3的表达.结果 PTPN3蛋白在50.9％的CRC组织中（57/112）下调,并与肿瘤TNM分期负相关（r=-0.253,P＜0.01）,PTPN3强表达的患者生存期有缩短的趋势（P＞0.05）.荧光素酶结果显示PTPN3是miR-95的靶基因,在CRC细胞株中过表达miR-95后,PTPN3 mRNA和蛋白表达均下降,而抑制miR-95后PTPN3表达增加.结论 PTPN3表达在CRC组织中下调,可能与肿瘤进展相关;miR-95调控是PTPN3在结直肠中表达下调的关键机制。
Objective To evaluate the expression and clinical significance of tyrosine-protein phosphatase non-receptor type 3 （PTPN3） expression in colorectal cancer （CRC） and to investigate the mechanism regulating PTPN3 expression.Methods Immunohistochemical staining was used to detect the protein expression of PTPN3 in 112 paired colorectal cancer and corresponding normal tissues.Luciferase assay was used to validate the regulation of microRNA-95 （miR-95） on PTPN3.The mRNA and protein expression of PTPN3 in CRC cells was examined by reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting,respectively.Results PTPN3 protein expression was down-regulated in 50.9% CRC tissues （57/112） as compared with their corresponding normal tissues,and negatively correlated with tumor TNM stage （r =-0.253,P 〈 0.01）,and patients with high PTPN3 expression showed a tendency for shortened survival time （P 〉 0.05）.Luciferase assay proved that PTPN3 was a target gene of miR-95.The mRNA and protein expression of PTPN3 was down-regulated in miR-95-overexpressed CRC cells,and up-regulated in miR-95-silenced CRC cells.Conclusion The down-regulation of PTPN3 in CRC tissues may be related with the progression of CRC.The miR-95 regulation is a key mechanism resulting in the down-regulation of PTPN3 in CRC.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第9期2210-2212,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金（81272299、81000867、81301784）
江苏省自然科学基金（BK20150004、BK20151108）
无锡市医管中心项目（YGZXZ1401）
