miR-548j-5p表达对胰腺癌侵袭能力的影响

doi:10.3877/cma.j.issn.2095-3232.2018.01.020

中华肝脏外科手术学电子杂志 ›› 2018, Vol. 07 ›› Issue (01) : 82 -86. doi: 10.3877/cma.j.issn.2095-3232.2018.01.020

所属专题：文献；

基础研究

miR-548j-5p表达对胰腺癌侵袭能力的影响

何睿智¹, 彭丰¹, 冯业晨¹, 郭兴军¹, 李旭¹, 王敏¹, 田锐¹, 朱峰¹, 秦仁义¹^,^†(

)

^1. 430030　武汉，华中科技大学同济医学院附属同济医院胆胰外科

收稿日期:2017-11-06 出版日期:2018-02-10
通信作者: 秦仁义
基金资助:
国家自然科学基金(81502633，81272659)

Effect of miR-548j-5p expression on invasion of pancreatic cancer

Ruizhi He¹, Feng Peng¹, Yechen Feng¹, Xingjun Guo¹, Xu Li¹, Min Wang¹, Rui Tian¹, Feng Zhu¹, Renyi Qin¹^,^†()

^1. Department of Biliary and Pancreatic Surgery, Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science & Technology, Wuhan 430030, China

Received:2017-11-06 Published:2018-02-10
Corresponding author: Renyi Qin
About author:
Corresponding author: Qin Renyi, Email: ryqin@tjh.tjmu.edu.cn

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目的

探讨miR-548j-5p在胰腺癌中的表达及其对胰腺癌侵袭能力的影响。

方法

本研究组织标本来源于2016年1月至2017年1月在华中科技大学同济医学院附属同济医院接受诊治的16例胰腺癌患者。其中男8例，女8例；年龄41～62岁，≤50岁7例，>50岁9例。患者均签署知情同意书，符合医学伦理学规定。采用RT-PCR检测胰腺癌及癌旁组织中miR-548j-5p的表达量。采用miR-548j-5p的mimic、inhibitor分别转染胰腺癌PANC-1细胞，构建过表达、低表达miR-548j-5p的PANC-1细胞，其对应的空载体Ctrl转染胰腺癌PANC-1细胞作为对照组。Transwell实验检测miR-548j-5p对PANC-1细胞侵袭能力的影响，划痕实验检测miR-548j-5p对PANC-1细胞迁移能力的影响。3组比较采用单因素方差分析和LSD-t检验。

结果

胰腺癌中miR-548j-5p相对表达量中位数为0.35（0.03～4.47），明显低于癌旁组织的4.30（0.04～65.55）（Z=-2.689，P<0.05）。Transwell实验结果显示miR-548j-5p过表达组的平均穿膜细胞数为（58±34）个，明显少于对照组的（231±61）个（LSD-t=-2.852，P<0.05）；miR-548j-5p低表达组穿膜细胞数为（491±149）个，明显多于对照组（LSD-t=4.313，P<0.05）。划痕实验结果显示miR-548j-5p过表达组划痕融合率为（20±8）%，明显低于对照组的（38±6）%（LSD-t=-3.759，P<0.05）；miR-548j-5p低表达组划痕融合率为（53±6）%，明显高于对照组的（38±6）%（LSD-t=2.946，P<0.05）。

结论

miR-548j-5p在胰腺癌中低表达，miR-548j-5p过表达能降低胰腺癌侵袭转移能力。

关键词: 微RNAS, 胰腺肿瘤, 肿瘤侵润, 肿瘤转移, 548j-5p基因

Objective

To explore the expression of miR-548j-5p in pancreatic cancer and its effect on the invasion of pancreatic cancer.

Methods

Tissue samples of 16 patients with pancreatic cancer who received treatment in Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science & Technology between January 2016 and January 2017 were collected. There were 8 males and 8 females, aged from 41 to 62, including 7 cases with an age of ≤50 and 9 cases with an age of >50. The informed consents of all patients were obtained and the local ethical committee approval was received. The expression of miR-548j-5p in pancreatic cancer and adjacent tissues was detected by RT-PCR. PANC-1 cells were transfected by mimic and inhibitor of miR-548j-5p to construct PANC-1 cells with over-expression and low-expression of miR-548j-5p. And in control group, PANC-1 cells were transfected with empty plasmid Ctrl. The effect of miR-548j-5p on PANC-1 invasion was detected by Transwell assay, and its effect on PANC-1 metastasis was evaluated by wound scratch assay. Comparison of three groups was conducted by one-way analysis of variance and LSD-t test.

Results

The median relative expression of miR-548j-5p in pancreatic cancer was 0.35(0.03-4.47), significantly lower than 4.30 (0.04-65.55) in adjacent tissues (Z=-2.689, P<0.05). Transwell assay results showed that the average number of cells penetrating membrane was (58±34) in miR-548j-5p over-expression group, significantly lower than (231±61) in control group (LSD-t=-2.852, P<0.05); and it was (491±149) in miR-548j-5p low-expression group, significantly higher than that in control group (LSD-t=4.313, P<0.05). The wound scratch assay results showed that the wound scratch healing rate was (20±8)% in miR-548j-5p over-expression group, significantly lower than (38±6)% in control group (LSD-t=-3.759, P<0.05); and it was (53±6)% in miR-548j-5p low-expression group, significantly higher than that in control group (LSD-t=2.946, P<0.05).

Conclusions

Low expression of miR-548j-5p is observed in pancreatic cancer. Over expression of miR-548j-5p can decrease the invasive and metastatic ability of pancreatic cancer.

Key words: MicroRNAs, Pancreatic neoplasms, Neoplasm invasiveness, Neoplasm metastasis, 548j-5p gene

图1 miR-548j-5p在胰腺癌组织中的表达情况

图2 Transwell法检测胰腺癌细胞侵袭能力

图3 细胞划痕实验检测胰腺癌细胞的迁移能力

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