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

Cancer Cell International

Yi-Hsiang Liu³  Yu-Hui Tseng²  Kee-Chin Sia²  Wei-Ting Chao²  Yung-Hsiang Hsu³  Yih-Shyong Lai⁴  Yen-Chang Clark Lai⁵  Chiung-Chi Cheng¹ 

[1] Center for General Education, Providence University, Taichung City, Taiwan;Department of Life Science, Tunghai University, Taichung City, Taiwan;Department of Pathology, Tzu Chi University, Hualien County, Taiwan;Department of Pathology, Chang Bing Show Chwan Memorial Hospital, No. 6, Lugong Rd., Lugang Town 505, Changhua County, Taiwan;Department of Pathology, Kaohsiung Medical University Hospital, Kaohsiung City, Taiwan

关键词: Plectin;    Hepatocellular carcinoma;    Focal adhesion kinase;    Cytoskeleton;    Cell motility;   

Others  :  1138458
DOI  :  10.1186/s12935-015-0177-1

 received in 2014-12-08, accepted in 2015-02-16,  发布年份 2015

【 摘 要 】

Background

Plectin 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 results

The 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.

Conclusion

Plectin-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.

【 授权许可】

   
2015 Cheng et al.; licensee BioMed Central.

【 图 表 】

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【 参考文献 】

• [1]Wiche G: Role of plectin in cytoskeleton organization and dynamics. J Cell Sci 1998, 111(Pt 17):2477-2486.
• [2]Burgstaller G, Gregor M, Winter L, Wiche G: Keeping the vimentin network under control: cell-matrix adhesion-associated plectin 1f affects cell shape and polarity of fibroblasts. Mol Biol Cell 2010, 21(19):3362-3375.
• [3]Osmanagic-Myers S, Wiche G: Plectin-RACK1 (receptor for activated C kinase 1) scaffolding: a novel mechanism to regulate protein kinase C activity. J Biol Chem 2004, 279(18):18701-18710.
• [4]Andrä K, Nikolic B, Stocher M, Drenckhahn D, Wiche G: Not just scaffolding: plectin regulates actin dynamics in cultured cells. Genes Dev 1998, 12(21):3442-3451.
• [5]Osmanagic-Myers S, Gregor M, Walko G, Burgstaller G, Reipert S, Wiche G: Plectin-controlled keratin cytoarchitecture affects MAP kinases involved in cellular stress response and migration. J Cell Biol 2006, 174(4):557-568.
• [6]Elliott CE, Becker B, Oehler S, Castanon MJ, Hauptmann R, Wiche G: Plectin transcript diversity: identification and tissue distribution of variants with distinct first coding exons and rodless isoforms. Genomics 1997, 42(1):115-125.
• [7]Garcia-Alvarez B, Bobkov A, Sonnenberg A, de Pereda JM: Structural and functional analysis of the actin binding domain of plectin suggests alternative mechanisms for binding to F-actin and integrin beta4. Structure 2003, 11(6):615-625.
• [8]Sevcik J, Urbanikova L, Kost’an J, Janda L, Wiche G: Actin-binding domain of mouse plectin. Crystal structure and binding to vimentin. Eur J Biochem 2004, 271(10):1873-1884.
• [9]Rezniczek GA, de Pereda JM, Reipert S, Wiche G: Linking integrin alpha6beta4-based cell adhesion to the intermediate filament cytoskeleton: direct interaction between the beta4 subunit and plectin at multiple molecular sites. J Cell Biol 1998, 141(1):209-225.
• [10]Kostan J, Gregor M, Walko G, Wiche G: Plectin isoform-dependent regulation of keratin-integrin alpha6beta4 anchorage via Ca2+/calmodulin. J Biol Chem 2009, 284(27):18525-18536.
• [11]Mierke CT: The role of focal adhesion kinase in the regulation of cellular mechanical properties. Phys Biol 2013, 10(6):065005.
• [12]Ritt M, Guan JL, Sivaramakrishnan S: Visualizing and manipulating focal adhesion kinase regulation in live cells. J Biol Chem 2013, 288(13):8875-8886.
• [13]Lightfoot HM Jr, Lark A, Livasy CA, Moore DT, Cowan D, Dressler L, et al.: Upregulation of focal adhesion kinase (FAK) expression in ductal carcinoma in situ (DCIS) is an early event in breast tumorigenesis. Breast Cancer Res Treat 2004, 88(2):109-116.
• [14]Oktay MH, Oktay K, Hamele-Bena D, Buyuk A, Koss LG: Focal adhesion kinase as a marker of malignant phenotype in breast and cervical carcinomas. Hum Pathol 2003, 34(3):240-245.
• [15]Van Eyken P, Desmet VJ: Cytokeratins and the liver. Liver 1993, 13(3):113-122.
• [16]Cheng CC, Liu YH, Ho CC, Chao WT, Pei RJ, Hsu YH, et al.: The influence of plectin deficiency on stability of cytokeratin18 in hepatocellular carcinoma. J Mol Histol 2008, 39(2):209-216.
• [17]Liu YH, Cheng CC, Ho CC, Chao WT, Pei RJ, Hsu YH, et al.: Plectin deficiency on cytoskeletal disorganization and transformation of human liver cells in vitro. Med Mol Morphol 2011, 44(1):21-26.
• [18]Roessler S, Long EL, Budhu A, Chen Y, Zhao X, Ji J, et al.: Integrative genomic identification of genes on 8p associated with hepatocellular carcinoma progression and patient survival. Gastroenterology 2012, 142(4):957-966. e912
• [19]Legate KR, Wickstrom SA, Fassler R: Genetic and cell biological analysis of integrin outside-in signaling. Genes Dev 2009, 23(4):397-418.
• [20]Hehlgans S, Haase M, Cordes N: Signalling via integrins: implications for cell survival and anticancer strategies. Biochim Biophys Acta 2007, 1775(1):163-180.
• [21]Webb DJ, Parsons JT, Horwitz AF: Adhesion assembly, disassembly and turnover in migrating cells – over and over and over again. Nat Cell Biol 2002, 4(4):E97-100.
• [22]Fraley SI, Feng Y, Krishnamurthy R, Kim DH, Celedon A, Longmore GD, et al.: A distinctive role for focal adhesion proteins in three-dimensional cell motility. Nat Cell Biol 2010, 12(6):598-604.
• [23]Ridley AJ: Rho GTPases and actin dynamics in membrane protrusions and vesicle trafficking. Trends Cell Biol 2006, 16(10):522-529.
• [24]Suetsugu S, Miki H, Takenawa T: Spatial and temporal regulation of actin polymerization for cytoskeleton formation through Arp2/3 complex and WASP/WAVE proteins. Cell Motil Cytoskeleton 2002, 51(3):113-122.