特异性下调LKB1的表达对肝癌细胞侵袭迁移能力的影响

沙亮; 何剑波; 李科志; 陈闯; 黄山; 赵荫农; 连芳; 邬国斌

doi:10.3969/j.issn.1001-5256.2017.05.016

特异性下调LKB1的表达对肝癌细胞侵袭迁移能力的影响

DOI: 10.3969/j.issn.1001-5256.2017.05.016

1. 广西医科大学附属肿瘤医院肝胆外科
2. 广西医科大学研究生院
3. 广西医科大学基础医学院

基金项目:

国家自然科学基金资助项目(81460426);国家自然科学基金资助项目(81360315)；

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中图分类号: R735.7
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出版历程
- 出版日期: 2017-05-20

Influence of specific down-regulation of LKB1 expression on invasion and migration of hepatoma cells

Department of Hepatobiliary Surgery, The Affiliated Tumor Hospital of Guangxi Medical University

Research funding:

摘要

摘要:
目的研究特异性下调LKB1的表达对肝癌细胞系SK-hep-1迁移侵袭能力的影响。方法 Real-time PCR和Western Blot检测人肝癌细胞株和人正常肝细胞HL7702中LKB1表达情况。设计并合成LKB1特异性双链小干扰RNA（LKB1-siRNA）,转染到高表达LKB1的人肝癌细胞株SK-hep-1中靶向沉默LKB1基因表达,并设置阴性对照组（NC组）。Real-time PCR和Western Blot检测LKB1基因的mRNA水平与蛋白表达水平的变化。利用CCK-8法检测肝癌细胞增殖能力。通过体外Transwell小室基质侵袭和迁移实验,观察LKB1表达沉默后对人肝癌细胞株SK-hep-1侵袭迁移能力的影响。计量资料两组间比较采用t检验,多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果与正常人肝细胞HL7702相比,LKB1基因和蛋白在人肝癌细胞SK-hep-1中高表达（P值均<0.05）。与NC组相比,siLKB1-1处理组SK-hep-1细胞中的LKB1基因和蛋白表达均明显降低（P值均<0.05）。与NC组相比,siLKB1-1转...
- 癌,肝细胞 /
- LKB1 /
- 细胞增殖 /
- 肿瘤侵润 /
- 细胞运动
Abstract:
Objective To investigate the influence of specific down-regulation of LKB1 expression on invasion and migration of the hepatoma cell line SK-hep-1.Methods Quantitative real-time PCR (RT-PCR) and Western blot were used to measure the expression of LKB1 in the human hepatoma cell line SK-hep-1 and normal hepatocytes (HL7702) .LKB1 specific double-strand small interfering RNA was designed, synthesized, and then transfected into the human hepatoma cell line SK-hep-1 with high expression of LKB1 to silence the expression of LKB1 gene.A negative control (NC) group was established.RT-PCR and Western blot were used to measure the changes in the mRNA and protein expression of LKB1.Cell Counting Kit-8 was used to measure the proliferative capacity of hepatoma cells.In vitro Transwell chamber invasion and migration assays were used to observe the influence of LKB1 gene silencing on invasion and migration of the human hepatoma cell line SK-hep-1.The t-test was used for comparison of continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between any two groups.Results Compared with normal human hepatocytes (HL7702) , SK-hep-1 cells had significantly higher mRNA and protein expression of LKB1 (both P<0.05) .Compared with the NC group, the siLKB1-1 treatment group had significant reductions in the mRNA and protein expression of LKB1 in SK-hep-1 cells (both P<0.05) .Compared with the NC group, the siLKB1-1 treatment group had a significantly higher cell proliferation rate and significant increases in invasion and migration (1.393 ±0.022 vs 1.1284±0.032, t=15.313, P<0.001;147±19 vs 83±21, t=14.879, P<0.001;113±13 vs 75±17, t=8.791, P<0.001) .Conclusion LKB1 may be involved in inhibiting the proliferation, invasion, and migration of hepatoma cells and can affect the prognosis of patients with liver cancer.
- carcinoma, hepatocellular /
- liver kinase b1 /
- cell proliferation /
- neoplasm invasiveness /
- cell movement

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