SFRP5基因沉默促进人类胰腺癌细胞系PANC-1增殖与迁移

基础医学与临床 ›› 2017, Vol. 37 ›› Issue (10): 1434-1439.

SFRP5基因沉默促进人类胰腺癌细胞系PANC-1增殖与迁移

刘保瑞¹,吴永娜²,王海平²,张辉²,潘建国¹,周文策²

1. 兰州大学
2. 兰州大学第一医院

收稿日期:2016-07-29 修回日期:2016-12-01 出版日期:2017-10-05 发布日期:2017-09-25
通讯作者: 周文策 E-mail:zhouwc129@163.com
基金资助:
中央高校基本科研业务费专项资金;甘肃省自然科学基金项目

SFRP5 gene silencing promotes proliferation and migration of human pancreatic cancer cell line PANC-1

Received:2016-07-29 Revised:2016-12-01 Online:2017-10-05 Published:2017-09-25

摘要/Abstract

摘要： 目的探讨慢病毒介导短发夹RNA( shRNA)沉默SFRP5基因对人类胰腺癌细胞系PANC-1细胞增殖、侵袭和迁移能力的影响。方法构建靶向SFRP5基因特异性shRNA慢病毒载体并转染人胰腺癌PANC-1细胞系，以空白质粒转染阴性对照组，未处理细胞做为空白对照组。用 real-time PCR及Western blot检测转染前后SFRP5 RNA以及蛋白的表达；CCK-8实验检测细胞体外增殖能力；使用Transwell小室实验分析细胞侵袭能力；细胞划痕实验分析细胞迁移能力。结果成功建立稳定转染shRNA-SFRP5胰腺癌PANC-1细胞株。SFRP5病毒转染组与阴性对照组及空白对照组相比细胞的增殖能力明显增加（P<0.01）; SFRP5病毒转染组的细胞侵袭、迁移能力明显高于阴性对照组及空白对照组（P<0.01）。结论 SFRP5慢病毒干扰载体能有效抑制SFRP5基因在人胰腺癌PANC-1细胞中的表达，进而促进细胞的增殖、侵袭和迁移。

关键词: [关键词] 慢病毒, 胰腺肿瘤, 生物学行为

Abstract: Objective To investigate the effect of lentivirus-mediated shRNA silencing SFRP5 on proliferation, invasion and migration of pancreatic cancer cell line PANC-1. Methods A SFRP5-knockdown recombinant plasmid was constructed and successfully transfected it into pancreatic cancer cell line PANC-1, blank plasmid transfection was treated as negative control and untreated cells as blank control group. The expression of SFRP5 at RNA and protein level in cell were detected by real-time PCR and Western blot, CCK8 assay was applied to examine the effect of SFRP5 silencing on the proliferation, the cell migration of pancreatic cancer cell line PANC-1 was analyzed by Transwell migration assay and cell scratch test was used to examine the cell invasion in PANC-1 cell. Results Stable transfected shRNA-SFRP5 cell of pancreatic cancer line was established successfully. The proliferation capacity of SFRP5 group was significantly higher compared to the negative control and blank control group by CCK8 assay(P<0.01).Similarly, cell invasion and migration of SFRP5 group was significantly higher compared to the negative control and blank control group(P<0.01). Conclusions SFRP5 lentiviral interference vectors can effectively decrease SFRP5 gene expression in PANC-1 cell of pancreatic cancer, thereby promoting cell proliferation, invasion and migration.

Key words: Key words: Lentiviral, Pancreatic neoplasms, Biological behavior

中图分类号:

R73-3

刘保瑞吴永娜王海平张辉潘建国周文策. SFRP5基因沉默促进人类胰腺癌细胞系PANC-1增殖与迁移[J]. 基础医学与临床, 2017, 37(10): 1434-1439.

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