Laminin-5 stimulates hepatocellular carcinoma growth through a different function of α6β4 and α3β1 integrins

Bergamini, Carlo; Sgarra, Concetta; Trerotoli, Paolo; Lupo, L.; Azzariti, Amalia; Antonaci, Salvatore; Giannelli, Gianluigi

doi:10.1002/hep.21936

Cited by 66 publications

(42 citation statements)

References 30 publications

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“…Cell proliferation was determined by MTT assay in accordance with other studies [11]. Briefly, 2 μg/cm 2 of LN (Sigma, St. Louis, MO, USA) was placed on a ten-dish plate and allowed to solidify at 37°C for two hours.…”

Section: Methodsmentioning

confidence: 99%

RNA interference targeting human integrin α6 suppresses the metastasis potential of hepatocellular carcinoma cells

Qiao

et al. 2013

Eur J Med Res

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BackgroundIncreased metastasis has been proved to be associated with a poor prognosis for hepatocellular carcinoma (HCC). There are higher-level expressions of integrin α6 in the tissues of HCC patients with a higher fatality rate. The aim of this study is to investigate the effect of short hairpin RNA (shRNA) silencing integrin α6 expression on the proliferation and metastasis in HCC cell lines.MethodsTwo human HCC cell lines, HepG2 and Bel-7402 were transfected with shRNA targeting human integrin α6. Protein and mRNA expression level were determined by western blot and real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) to detect the transfected efficacy. The metastasis potential of HCC cells was evaluated by their proliferation, adhesion and invasion abilities. Cell proliferation was measured by MTT assay. Adhesion ability was measured by adhesion and spreading assays. The expression of matrix metalloproteinases (MMPs) was measured by qRT-PCR. The potential of invasion was measured by qRT-PCR and Transwell chamber assay. PI3K inhibitor LY294002 was used to explore the signal pathways of integrin α6 in HCC cells.ResultsWestern blot and qRT-PCR detection showed that over 75% of integrin α6 expression in HCC cells was through knockdown by shRNA. Proliferation, adhesion, spreading and invasion of HepG2 and Bel-7402 cells were dramatically decreased in cells transfected with shRNA compared to the control cells. P-ERK and p-AKT were reduced by shRNA targeting integrin α6 and PI3K inhibitor LY294002.ConclusionKnockdown integrin α6 can inhibit the proliferation and metastasis of HCC cells through PI3K/ARK and MAPK/ERK signal pathways by shRNA in vitro. Integrin α6 can mediate the metastasis potential, and can be used as a candidate target for therapy in HCC resulting in improved patients’ survival.

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Section: Methodsmentioning

confidence: 99%

RNA interference targeting human integrin α6 suppresses the metastasis potential of hepatocellular carcinoma cells

Qiao

et al. 2013

Eur J Med Res

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“…Cell lysates were prepared directly in cell extraction buffer (Biosource International) supplemented with a protease inhibitor cocktail (Sigma), as described previously [12]. CM was concentrated (Centricon; Millipore).…”

Section: Western Blot Analysismentioning

confidence: 99%

Hepatic stellate cells stimulate HCC cell migration via laminin-5 production

et al. 2011

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Activated HSCs (hepatic stellate cells) are the main source of extracellular matrix proteins present in cirrhotic liver on which HCC (hepatocellular carcinoma) commonly develops. HCC cells behave differently according to differences in the surrounding microenvironment. In the present study, we have investigated a mechanism whereby HSCs modulate the migratory activity of HCC cells. We used primary cultures of human HSCs to investigate their effect on Hep3B, Alexander, HLE and HLF HCC cells. The expression of Ln-5 (laminin-5) was documented at transcript and protein levels both in vitro and in vivo. HCC cells strongly adhere, migrate and spread in the presence of HSC-conditioned medium and of co-culture. HSCs produce and secrete Ln-5 in the CM (conditioned medium). The electrophoretic pattern of secreted Ln-5 is consistent with that of a migratory substrate, showing the presence of the γ2x fragment. Blocking antibodies against Ln-5 inhibit HCC migration in the presence of HSC-CM. HCC cells migrate very poorly in the presence of Ln-5 immunodepleted HSC-CM. HCC migration in the presence of HSCs is dependent on the MEK [MAPK (mitogen-activated protein kinase)/ERK (extracellular-signal-regulated kinase) kinase]/ERK pathway, but not the PI3K (phosphoinositide 3-kinase)/Akt pathway. HSC-CM, as well as Ln-5, activates the MEK/ERK but not the PI3K/Akt pathway. In human HCC tissues, Ln-5 is mainly distributed along α-SMA (smooth muscle actin)-positive cells, whereas in peritumoural tissues, Ln-5 is absent. HSCs stimulate HCC migration via the production and secretion of Ln-5.

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“…The overexpression of β1 integrin inhibits HCC cell proliferation by preventing Skp-2 dependent degradation of p27 via PI3K pathways (135). On the other hand, the overexpression of α3β1 and α6β4 integrin is associated with increased migration and invasion of HCC cells in an Ln-5 dependent condition (136)(137)(138)(139).…”

Section: Hepatic Stellate Cells (Hscs)mentioning

confidence: 99%

The tumor microenvironment in hepatocellular carcinoma (Review)

et al. 2012

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Abstract. The tumor microenvironment has been largely studied as a dynamic system orchestrated by inflammatory cells, including cancer cells, stroma as well as the extracellular matrix. It is useful to describe and predict the phenotypic characteristics of cancer. Furthermore, a better understanding of its interplay with the various aspects of the tumor cells may be utilized for the discovery of novel molecular targets. Liver cancer is considered a model of the relation occurring between the tumor microenvironment and tumor development. The chronic inflammatory status of the liver, sustained by the infection of hepatitis viruses, as well as the production of cytokines and growth factors within the parenchyma, lead to an intricate microenvironment. The identification of novel molecular therapeutic targets may improve the outcome of patients with liver cancer as it remains the third leading cause of cancer death worldwide. In the present study, the tumor microenvironment in hepatocellular carcinoma (HCC) was explored by a review of the literature. Studies on hepatitis virus infections and the consequent chronic inflammatory status were examined. In this context, immune-mediated and/or virusrelated molecular mechanisms have been hypothesized as being responsible for liver cancer development. The interlink among HCC microenvironment components, comprising cellular elements, cytokines, growth factors and several proteins is also described together with the role of matrix metalloproteinases in HCC development. Finally, the rationale for targeting tumorstromal interface is summarized in the context of new therapeutic opportunities.

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Laminin-5 stimulates hepatocellular carcinoma growth through a different function of α6β4 and α3β1 integrins

Cited by 66 publications

References 30 publications

RNA interference targeting human integrin α6 suppresses the metastasis potential of hepatocellular carcinoma cells

RNA interference targeting human integrin α6 suppresses the metastasis potential of hepatocellular carcinoma cells

Hepatic stellate cells stimulate HCC cell migration via laminin-5 production

The tumor microenvironment in hepatocellular carcinoma (Review)

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