Transient knockdown-mediated deficiency in plectin alters hepatocellular motility in association with activated FAK and Rac1-GTPaseReport as inadecuate




Transient knockdown-mediated deficiency in plectin alters hepatocellular motility in association with activated FAK and Rac1-GTPase - Download this document for free, or read online. Document in PDF available to download.

Cancer Cell International

, 15:29

First Online: 07 March 2015Received: 08 December 2014Accepted: 16 February 2015

Abstract

BackgroundPlectin is one of the cytolinker proteins that play a crucial role in maintaining the integrity of cellular architecture. It is a component of desmosome complexes connecting cytoskeletal proteins and trans-membrane molecules. In epithelial cells, plectin connects cytokeratins and integrin α6β4 in hemidesmosomes anchoring to the extracellular matrix. In addition to the function of molecular adherent, plectin has been reported to exhibit functions affecting cellular signals and responsive activities mediated by stress, cellular migration, polarization as well as the dynamic movement of actin filaments. Plectin deficiency in hepatocellular carcinoma results in abnormal expression of cytokeratin 18 and disassembled hemidesmosome. Therefore, it is hypothesized that the plectin deficiency-mediated collapse of cytoskeleton may modulate cellular motility that is associated with consequent metastatic behaviors of cancer cells.

Methods and resultsThe cellular motility of plectin-deficient Chang liver cells generated by transient knockdown were analyzed by trans-well migration assay and the results revealed a higher migration rate. The confocal microscopy also demonstrated less organized and more polarized morphology as well as more focal adhesion kinase activity in comparison with that of the mock Chang liver cells. Furthermore, plectin-knockdown in Chang liver cells was associated with a higher activity of Rac1-GTPase in accordance with the results of the Rac1 pull-down assay. The immunohistochemical assay on human hepatocellular carcinoma showed that the expression of focal adhesion kinase was increased in the invasive front of tumor.

ConclusionPlectin-deficient human hepatic cells exhibit higher cell motility associated with increase in focal adhesion kinase activity that are comparable to the properties of invasive hepatocellular carcinoma.

KeywordsCell motility Cytoskeleton Focal adhesion kinase Hepatocellular carcinoma Plectin AbbreviationsCK18Cytokeratin 18

CKsCytokeratins

FAKFocal adhesion kinase

FAsFocal adhesions

FnIIIFibronectin type III

GSTGlutathione S-transferase

HCCHepatocellular carcinoma

HDHemidesmosome

IFsIntermediate filaments

MAPMitogen-activated protein

MFsMicrofilaments

MTsMicrotubules

Pakp21-activated kinase

PBSPhosphate buffered saline

PBSTPhosphate buffered saline with Tween-20

PKCProtein kinase C

PMSFPhenylmethylsulfonylfluoride

Rac1Ras-related C3 botulinum toxin substrate 1

RACK1Receptor activated C kinase 1

siRNASmall interfering RNA

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Author: Chiung-Chi Cheng - Yen-Chang Clark Lai - Yih-Shyong Lai - Yung-Hsiang Hsu - Wei-Ting Chao - Kee-Chin Sia - Yu-Hui Tseng -

Source: https://link.springer.com/article/10.1186/s12935-015-0177-1



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