基础研究(Basic Research)

基质硬度对PKN3表达及肝细胞癌侵袭转移的影响

Published at: 2016年第25卷第10期

欧阳锡武 1 , 陶一明 1 , 王志明 1
1 中南大学湘雅医院 肝脏外科,湖南 长沙410008
通讯作者 志明 王 Email: wzmxycsu@hotmail.com
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2016.10.015
基金:
国家自然科学基金面上资助项目 81372630,81372631
湖南自然科学基金资助项目 12JJ3118
湖南省科学技术厅科技计划资助项目 2011SK3228

摘要

目的:探讨蛋白激酶C相关激酶N3(PKN3)表达与肝细胞癌(HCC)细胞侵袭转移的关系及机制。方法:检测72例出血坏死表型HCC(HN-HCC)与32例无出血坏死表型HCC(NHN-HCC)组织标本中胶原纤维含量与PKN3的表达;将肝癌HCCLM3细胞在不同硬度的聚丙烯酰胺水凝胶培养后,检测HCCLM3细胞的运动侵袭能力、PKN3基因与蛋白的表达,以及RhoC活性。通过PKN3干扰以及PKN3干扰同时加RhoC过表达,观察HCCLM3细胞运动侵袭能力以及FAK、ROCK2、E-cadherin、Fibronectin蛋白表达的变化。结果:与NHN-HCC组织比较,HN-HCC组织胶原纤维含量与PKN3蛋白表达明显增加(均P<0.05);与软基质中培养的HCCLM3细胞比较,在硬基质中培养的HCCLM3细胞PKN3的基因与蛋白表达明显增加、侵袭运动能力明显增强、RhoC活性明显升高(均P<0.05),后者活性随PKN3表达的抑制而抑制。与对照组细胞比较,干扰PKN3表达后,HCCLM3细胞运动侵袭能力明显降低,FAK、ROCK2、Fibronectin蛋白表达明显下调,E-cadherin蛋白表达明显下调(均P<0.05),而PKN3干扰同时增加RhoC表达,HCCLM3细胞以上变化不明显(均P>0.05)。结论:基质硬度增加能上调PKN3-ROCK2信号通路活性,从而促进HCC侵袭转移,该机制可能是HN-HCC侵袭转移能力较高的重要原因。


Influence of matrix stiffness on PKN3 expression and invasion/metastasis in hepatocellular carcinoma

Abstract

Objective: To investigate the relationship between expression of PKC-related kinase N3 (PKN3) and the invasion and metastasis of hepatocellular carcinoma (HCC) cells and the mechanism. Methods: The collagen fiber content and PKN3 expression in the specimens of 72 HCC tissues with hemorrhagic/necrotic phenotype (HN-HCC) and 32 HCC tissues without hemorrhagic/necrotic phenotype (NHN-HCC) were determined. HCC HCCLM3 cells were cultured in polyacrylamide hydrophilic gels of different stiffness, and then, the migration and invasion abilities and the PKN3 gene and protein expressions as well as RhoC activity were measured. After PKN3 interference or PKN3 interference with concomitant RhoC overexpression, the changes in migration and invasion abilities and protein expressions of ROCK2, E-cadherin, Fibronectin in HCCLM3 cells were analyzed. Results: Compared with NHN-HCC tissue, collagen fiber content and PKN3 protein expression were significantly increased in HN-HCC (both P<0.05); in HCCLM3 cells cultured in firm matrix compared with HCCLM3 cells cultured in soft matrix, both gene and protein expressions of PKN3, and the migration and invasion abilities as well as the RhoC activity were significantly increased (all P<0.05), while the RhoC activity was suppressed with inhibition of PKN3 expression. Compared with control HCCLM3 cells, the migration and invasion abilities and the protein expressions of FAK, ROCK2 and Fibronectin were significantly decreased, and E-cadherin protein expression was significantly increased in HCCLM3 cells after PKN3 interference (all P<0.05), while the above changes were not obvious in HCCLM3 cells after PKN3 interference with concomitant RhoC overexpression (all P>0.05). Conclusion: Increased matrix stiffness can up-regulate the activity of PKN3-ROCK2 signaling pathway, and thereby promote the invasion and metastasis of HCC, which may be an important mechanism for the high invasion and metastasis ability of HN-HCC.


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引用

引用本文: 锡武 欧阳, 一明 陶, 志明 王. 基质硬度对PKN3表达及肝细胞癌侵袭转移的影响[J]. 中国普通外科杂志, 2016, 25(10): 1455-1460.
Cite this article as: OUYANG Xiwu, TAO Yiming, WANG Zhiming . Influence of matrix stiffness on PKN3 expression and invasion/metastasis in hepatocellular carcinoma [J]. Chin J Gen Surg, 2016, 25(10): 1455-1460.