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

Cheng CC, Lai YC, Lai YS, Hsu YH, Chao WT, Sia KC, Tseng YH, Liu YH - Cancer Cell Int. (2015)

Bottom Line: 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.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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Chang Bing Show Chwan Memorial Hospital, No. 6, Lugong Rd., Lugang Town, Changhua County 505 Taiwan ; Center for General Education, Providence University, Taichung City, Taiwan.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus

Plectin depletion enhanced Chang liver cell migration. Plectin siRNA-transfected Chang liver cells were transferred to trans-well migration insert, followed by 4 hour incubation as described in the section of “materials and methods”. Plectin-knockdown Chang liver cell (refer to as Plectin siRNA) significantly showed a higher level of cell migration. Data are presented as mean ± standard error obtained from three independent experiments, *** indicates p < 0.001. The migrated cell number of mock group was assumed as 100%. Cell migration of plectin-knockdown Chang liver cell is presented as the ratio to that of the mock group.
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Fig1: Plectin depletion enhanced Chang liver cell migration. Plectin siRNA-transfected Chang liver cells were transferred to trans-well migration insert, followed by 4 hour incubation as described in the section of “materials and methods”. Plectin-knockdown Chang liver cell (refer to as Plectin siRNA) significantly showed a higher level of cell migration. Data are presented as mean ± standard error obtained from three independent experiments, *** indicates p < 0.001. The migrated cell number of mock group was assumed as 100%. Cell migration of plectin-knockdown Chang liver cell is presented as the ratio to that of the mock group.

Mentions: The results of trans-well migration assay demonstrated that plectin-deficiency in Chang liver cells derived from transient knockdown revealed a significant higher rate of cell migration (about 300%, p < 0.001) compared with that of the mock counterpart (Figure 1).Figure 1


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

Cheng CC, Lai YC, Lai YS, Hsu YH, Chao WT, Sia KC, Tseng YH, Liu YH - Cancer Cell Int. (2015)

Plectin depletion enhanced Chang liver cell migration. Plectin siRNA-transfected Chang liver cells were transferred to trans-well migration insert, followed by 4 hour incubation as described in the section of “materials and methods”. Plectin-knockdown Chang liver cell (refer to as Plectin siRNA) significantly showed a higher level of cell migration. Data are presented as mean ± standard error obtained from three independent experiments, *** indicates p < 0.001. The migrated cell number of mock group was assumed as 100%. Cell migration of plectin-knockdown Chang liver cell is presented as the ratio to that of the mock group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4358909&req=5

Fig1: Plectin depletion enhanced Chang liver cell migration. Plectin siRNA-transfected Chang liver cells were transferred to trans-well migration insert, followed by 4 hour incubation as described in the section of “materials and methods”. Plectin-knockdown Chang liver cell (refer to as Plectin siRNA) significantly showed a higher level of cell migration. Data are presented as mean ± standard error obtained from three independent experiments, *** indicates p < 0.001. The migrated cell number of mock group was assumed as 100%. Cell migration of plectin-knockdown Chang liver cell is presented as the ratio to that of the mock group.
Mentions: The results of trans-well migration assay demonstrated that plectin-deficiency in Chang liver cells derived from transient knockdown revealed a significant higher rate of cell migration (about 300%, p < 0.001) compared with that of the mock counterpart (Figure 1).Figure 1

Bottom Line: 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.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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Chang Bing Show Chwan Memorial Hospital, No. 6, Lugong Rd., Lugang Town, Changhua County 505 Taiwan ; Center for General Education, Providence University, Taichung City, Taiwan.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus