Plectin Promotes the Migration of Hepatocellular Carcinoma Cells Through Inducing F-actin Polymerization

XU Rushuang, YANG Lingxiao, SONG Guanbin

Abstract

To explore the relationship between the expression of plectin and the migration of hepatocellular carcinoma (HCC) cells and to elucidate the molecular mechanisms by which plectin expression affects the migration of HCC cells.Methods  First of all, Western blot was performed to determine the expression of plectin in normal hepatocytes and HCC cells. Secondly, a plectin-downregulated HCC cell strain was established and the control group (shNC group) and shPLEC group were set up. Each group was divided into a vehicle control group (shNC+DMSO group or shPLEC+DMSO group) and a F-actin cytoskeleton polymerization inducer Jasplakinolide group (shNC+Jasp group or shPLEC+Jasp group). Western blot was performed to determine the expression of plectin and epithelial-mesenchymal transition (EMT)-related proteins, including N-cadherin, vimentin, and E-cadherin. HCC cell migration was evaluated by Transwell assay. KEGG (Kyoto Encyclopedia of Genes and Genomes) was used to analyze the signaling pathways related to plectin gene. The polymerization of F-actin was analyzed by immunofluorescence assay.Results  Compared with the normal hepatocytes, HCC cells showed high expression of plectin. Compared with those in the shNC group, the expression of plectin in the shPLEC group was decreased (P<0.05), the migration ability of HCC cells was weakened (P<0.05), and the EMT process was inhibited (with the expression of N-cadherin and vimentin being decreased and the expression of E-cadherin being increased) (P<0.05). KEGG analysis showed that the regulation of cytoskeletal F-actin was most closely associated with plectin and cytoskeletal F-actin depolymerized in the shPLEC group. After treatment with Jasplakinolide, an inducer of F-actin cytoskeleton polymerization, the migration ability of HCC cells in the shPLEC+Jasp group was enhanced compared with that of shPLEC+DMSO group (P<0.05) and the EMT process was restored (with the expression of N-cadherin and vimentin being increased and the expression of E-cadherin being decreased) (P<0.05). In addition, the polymerization of cytoskeletal F-actin in HCC cells was also restored.Conclusion  Plectin is highly expressed in HCC cells. Plectin promotes the migration and the EMT of HCC cells through inducing F-actin polymerization.


Keywords: Plectin,  Hepatocellular carcinoma cells,  Cell migration assays, Epithelial-mesenchymal transition,  Cytoskeleton,  F-actin  

Full Text:

PDF


References


SUNG H， FERLAY J， SIEGEL R L， et al. Global cancer statistics 2020: globocan estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin，2021，71(3): 209–249. doi: 10.3322/caac. 21660.

HE Y， LUO L， SHAN R， et al. Development and validation of a nomogram for predicting postoperative early relapse and survival in hepatocellular carcinoma. J Natl Compr Canc Netw，2023: 1–7. doi: 10. 6004/jnccn.2023.7069.

LU L， ZHAN M， LI X Y， et al. Clinically approved combination immunotherapy: Current status, limitations, and future perspective. Curr Res Immunol，2022，3: 118–127. doi: 10.1016/j.crimmu.2022.05.003.

MU M， HUANG C X， QU C， et al. Targeting ferroptosis-elicited inflammation suppresses hepatocellular carcinoma metastasis and enhances sorafenib efficacy. Cancer Res，2024. doi: 10.1158/0008-5472. CAN-23-1796.

XIAO Y， LI Y， SHI D， et al. MEX3C-mediated decay of SOCS3 mRNA promotes JAK2/STAT3 signaling to facilitate metastasis in hepatocellular carcinoma. Cancer Res，2022，82(22): 4191–4205. doi: 10.1158/0008-5472. CAN-22-1203.

MA J， ZENG S， ZHANG Y， et al. BMP4 promotes oxaliplatin resistance by an induction of epithelial-mesenchymal transition via MEK1/ERK/ELK1 signaling in hepatocellular carcinoma. Cancer Lett， 2017，411: 117–129. doi: 10.1016/j.canlet.2017.09.041.

RICE S J， TSELEPI M， SORIAL A K， et al. Prioritization of PLEC and GRINA as osteoarthritis risk genes through the identification and characterization of novel methylation quantitative trait loci. Arthritis Rheumatol，2019，71(8): 1285–1296. doi: 10.1002/art.40849.

FAVRE B， SCHNEIDER Y， LINGASAMY P， et al. Plectin interacts with the rod domain of type Ⅲ intermediate filament proteins desmin and vimentin. Eur J Cell Biol，2011，90(5): 390–400. doi: 10.1016/j.ejcb.2010. 11.013.

SUTOH YONEYAMA M， HATAKEYAMA S， HABUCHI T， et al. vimentin intermediate filament and Plectin provide a scaffold for invadopodia, facilitating cancer cell invasion and extravasation for metastasis. Eur J Cell Biol，2014，93(4): 157–169. doi: 10.1016/j.ejcb.2014. 03.002.

KULKARNI T， MUKHOPADHYAY D， BHATTACHARYA S. Dynamic alteration of poroelastic attributes as determinant membrane nanorheology for endocytosis of organ specific targeted gold nanoparticles. J Nanobiotechnol，2022，20(1): 74. doi: 10.1186/s12951-大学, 2016.

SONG L L. Investigation metastasis-associated proteins in heptocellular carcinoma based on iTRAQ. Chongqing: Chongqing Medical University， 2016.

INFANT E， ETIENNE-MANNEVILLE S. Intermediate filaments: integration of cell mechanical properties during migration. Front Cell Dev Biol，2022，10: 951816. doi: 10.3389/fcell.2022.951816.

BERA K， KIEPAS A， ZHANG Y， et al. The interplay between physical cues and mechanosensitive ion channels in cancer metastasis. Front Cell Dev Biol，2022，10: 954099. doi: 10.3389/fcell.2022.954099.

WINTER L， KUZNETSOV A V， GRIMM M， et al. Plectin isoform P1b and P1d deficiencies differentially affect mitochondrial morphology and function in skeletal muscle. Hum Mol Genet，2015，24(16): 4530–4544. doi: 10.1093/hmg/ddv184.

FUCHS P， ZÖRER M， REZNICZEK G A， et al. Unusual 5' transcript complexity of Plectin isoforms: novel tissue-specific exons modulate actin binding activity. Hum Mol Genet，1999，8(13): 2461–2472. doi: 10.1093/hmg/8.13.2461.

MOCH M， WINDOFFER R， SCHWARZ N， et al. Effects of Plectin depletion on keratin network dynamics and organization. PLoS One， 2016，11(3): e0149106. doi: 10.1371/journal.pone.0149106.

OSMANAGIC S， GREGOR M， WALKO G， et al. Plectin-controlled keratin cytoarchitecture affects MAP kinases involved in cellular stress response and migration. J Cell Biol，2006，174(4): 557–568. doi: 10.1083/jcb.200605172.

KUBURICH N A， Den HOLLANDER P， CASTANEDA M， et al. Stabilizing vimentin phosphorylation inhibits stem-like cell properties and metastasis of hybrid epithelial/mesenchymal carcinomas. Cell Rep， 2023，42(12): 113470. doi: 10.1016/j.celrep.2023.113470.

CASTAÑÓN M J， WALKO G， WINTER L， et al. Plectin-intermediate filament partnership in skin, skeletal muscle, and peripheral nerve. Histochem Cell Biol，2013，140(1): 33–53. doi: 10.1007/s00418-013-1102-0.

TRAN V D， KUMAR S. Transduction of cell and matrix geometric cues by the actin cytoskeleton. Curr Opin Cell Biol，2021，68: 64–71. doi: 10. 1016/j.ceb.2020.08.016.

BONAKDAR N， SCHILLING A， SPÖRRER M， et al. Determining the mechanical properties of Plectin in mouse myoblasts and keratinocytes. Exp Cell Res，2015，331(2): 331–337. doi: 10.1016/j.yexcr.2014.10.001. ANDRÄ K， NIKOLIC B， STÖCHER M， et al. Not just scaffolding: Plectin regulates actin dynamics in cultured cells. Genes Dev，1998， 12(21): 3442–3451. doi: 10.1101/gad.12.21.3442.

SUN Y C， LI H， LUO Q， et al. Mechanical heterogeneity of tumor tissues and epithelial-mesenchymal transition of tumor cells. J Med Biomech，2021，36(4): 658–663. doi: 10.16156/j.1004-7220.2021.04.024.

ZHONG X， LONG H， CHEN L， et al. Stiffness on shear wave elastography as a potential microenvironment biomarker for predicting tumor recurrence in HBV-related hepatocellular carcinoma. Insights Imaging，2023，14(1): 147. doi: 10.1186/s13244-023-01505-7.

SUN Y， LI H， CHEN Q， et al. The distribution of liver cancer stem cells correlates with the mechanical heterogeneity of liver cancer tissue. Histochem Cell Biol，2021，156(1): 47–58. doi: 10.1007/s00418-021-01979-w.


Refbacks

  • There are currently no refbacks.