Integrin-Linked Kinase Overexpression and Its Oncogenic Role in Promoting Tumorigenicity of Hepatocellular Carcinoma

Background: Integrin-linked kinase (ILK) was first discovered as an integrin β1-subunit binding protein. It localizes at the focal adhesions and is involved in cytoskeleton remodeling. ILK overexpression and its dysregulated signaling cascades have been reported in many human cancers. Aberrant expression of ILK influenced a wide range of signaling pathways and cellular functions. Although ILK has been well characterized in many malignancies, its role in hepatocellular carcinoma (HCC) is still largely unknown. Methodology/Principal Findings: Quantitative PCR analysis was used to examine ILK mRNA expression in HCC clinical samples. It was shown that ILK was overexpressed in 36.9% (21/57) of HCC tissues when compared to the corresponding non-tumorous livers. The overall ILK expression level was significantly higher in tumorous tissues (P = 0.004), with a significant stepwise increase in expression level along tumor progression from tumor stage I to IV (P = 0.045). ILK knockdown stable clones were established in two HCC cell lines, BEL7402 and HLE, and were subjected to different functional assays. Knockdown of ILK significantly suppressed HCC cell growth, motility and invasion in vitro and inhibited tumorigenicity in vivo. Western blot analysis revealed a reduced phosphorylated-Akt (pAkt) at Serine-473 expression in ILK knockdown stable clones when compared to control clones. Conclusion/Significance: This study provides evidence about the clinical relevance of ILK in hepatocarcinogenesis. ILK was found to be progressively elevated along HCC progression. Here our findings also provide the first validation about the oncogenic capacity of ILK in vivo by suppressing its expression in HCC cells. The oncogenic role of ILK is implicated to be mediated by Akt pathway. © 2011 Chan et al.published_or_final_versio

A study of the signal transduction pathway ILK/p-Akt in human hepatocellular carcinoma: correlation with the expression of factors implicated in apoptosis and cellular proliferation

The hepatic carcinogenesis is a many phased process that leads progressively to the malignant transformation of hepatic cell via various molecular mechanisms. Recently, the enormous importance of the tumor microenvironment for the growth and development of cancer, i.e. the interaction between cancer cells and the extacellular matrix, has been proven. Particular interest is presented in the interaction of tumor cells with the extracellular matrix via integrins, which appears to be involved in all the stages of carcinogenesis. An important molecule in the aforementioned process is the ILK (Integrin-Linked Kinase), a serine-threonine kinase that is implicated in the signal transduction pathways that begin from integrines, growth factors and participate in the regulation of nodal for the tumor cell functions, such as control of cell cycle, loss of cell adhesion structures, suppression of apoptosis and activation of angiogenesis. It has been realized that the signal transduction pathway ILK /p-Akt plays a nodal role so much in the biology of cirrhosis as well as in hepatocellular cancer and probably constitutes a molecular connection between the two conditions, confirming the formulated theory of axis of chronic inflammation-cirrhosis-cancer. Also in the pathogenicity of cirrhosis and liver cancer, it appears that the epithelial to mesenchymal transition (EMT) phenomenon plays a role. This is proven by the loss of expression of E-cadherin and the transport of b-catenin in the nucleus that leads to loss of cellular connections from the membrane as well as from the overexpression of molecules ILK and p-Akt. Finally, the overexpression of survivin and cyclin-D1 in cirrhosis and hepatocellular carcinoma show the importance of antiapoptotic activity and increased cellular proliferation in the particular disease entities.Η ηπατική καρκινογένεση είναι μια πολυσταδιακή διαδικασία που οδηγεί προοδευτικά στον κακοήθη μετασχηματισμό του ηπατικού κυττάρου, μέσω ποικίλων μοριακών μηχανισμών. Πρόσφατα αναδεικνύεται ολοένα και σε μεγαλύτερο βαθμό, η τεράστια σημασία που έχει για την ανάπτυξη και εξέλιξη του καρκίνου το μικροπεριβάλλον του όγκου, η αλληλεπίδραση δηλαδή των καρκινικών κυττάρων με την εξωκυττάρια ουσία. Ιδιαίτερο ενδιαφέρον παρουσιάζει η αλληλεπίδραση των καρκινικών κυττάρων με την εξωκυττάρια ουσία μέσω των ιντεγκρινών, η οποία φαίνεται ότι εμπλέκεται σε όλα τα στάδια καρκινογένεσης. Σημαντικό μόριο στην προαναφερθείσα διαδικασία αποτελεί η ILK (Integrin-Linked Kinase), μία κινάση σερίνης-θρεονίνης που παρεμβάλλεται στα σηματοδοτικά μονοπάτια που ξεκινούν από ιντεγκρίνες, αυξητικούς παράγοντες και συμμετέχει στη ρύθμιση κομβικών για το καρκινικό κύτταρο λειτουργιών όπως έλεγχο του κυτταρικού κύκλου, απώλεια των δομών συνοχής του κυττάρου, αναστολή της απόπτωσης και ενεργοποίηση της αγγειογένεσης. Διαπιστώθηκε πως το σηματοδοτικό μονοπάτι ILK/p-Akt διαδραματίζει κομβικό ρόλο τόσο στη βιολογία της κίρρωσης όσο και του ηπατοκυτταρικού καρκίνου και πιθανόν αποτελεί μοριακό σύνδεσμο μεταξύ των δύο καταστάσεων, επιβεβαιώνοντας τη διατυπωμένη θεωρία περί άξονα χρόνιας φλεγμονής-ίνωσης (κίρρωσης)-καρκίνου. Επίσης στην παθογένεια της κίρρωσης και του καρκίνου ήπατος, φαίνεται πως ρόλο έχει το φαινόμενο της επιθηλιακής προς μεσεγχυματική μετατροπή (EMT-epithelial to mesenchymal transition). Αυτό αποδεικνύεται από την απώλεια έκφρασης της E-καντχερίνης και τη μεταφορά της β-κατενίνης στον πυρήνα που οδηγούν σε απώλεια των κυτταρικών συνδέσεων από τη μεμβράνη καθώς και από την υπερέκφραση των μορίων ILK και p-Akt. Τέλος η υπερέκφραση survivin και κυκλίνης-D1 στην κίρρωση και στο ηπακυτταρικό καρκίνωμα καταδεικνύουν τη σημασία της αντιαποπτωτικής δραστηριότητας και του αυξημένου κυτταρικού πολλαπλασιασμού στις συγκεκριμένες νοσολογικές οντότητες

Survivin overexpression in HCC and liver cirrhosis differentially correlates with p-STAT3 and E-cadherin

Survivin, a member of the family of inhibitorof apoptosis proteins, functions as a key regulator ofapoptosis and cell proliferation. Overexpression ofsurvivin has been implicated in several human cancers,including human hepatocellular carcinoma (HCC).Although several factors have been shown in vitrotoupregulate survivin expression in cancer cells, the invivoregulators of survivin in human hepato-carcinogenesis are largely unknown. We studied byimmunohistochemistry the protein expression ofsurvivin in relation to cyclin D1, phosphorylated signaltransducer and activator of transcription 3 (p-STAT3), ß-catenin, E-cadherin and phosphorylated-Akt (p-Akt) in69 cases of HCC and adjacent liver cirrhosis. Survivinwas expressed in 63/69 (91.3%) cases of HCC and in40/47 (85.1%) cases of liver cirrhosis. Survivinlocalization in HCC was exclusively nuclear, whileintense cytoplasmic and low nuclear expression ofsurvivin was observed in cases of cirrhosis. Survivinexpression in HCC correlated significantly with lowgrade tumors, expression of cyclin D1 and p-STAT3.Expression of survivin in liver cirrhosis correlated withdownregulation of E-cadherin expression. There was nosignificant correlation of survivin with ß-catenin or p-Akt in HCC or liver cirrhosis. In conclusion, we showedan association of nuclear survivin with welldifferentiated HCC, as well as with the expression of thecell cycle regulator cyclin D1. Activation of STAT3 andloss of E-cadherin but not ß-catenin or Akt pathwaysseem to be implicated in survivin upregulation in HCCand liver cirrhosis

Lapatinib-induced hepatitis: A case report

Lapatinib is an inhibitor of the tyrosine kinases of human epidermal growth factor receptor type 2 (HER2) and epidermal growth factor receptor type 1, with clinical activity in HER2-positive metastatic breast cancer. We present here a 60 year-old patient with metastatic breast cancer who presented with jaundice and increased serum aminotransferase levels and who had been treated with lapatinib for the previous 14 days. Laboratory tests excluded other causes of acute liver injury. Liver biopsy revealed lesions compatible with drug-induced hepatotoxicity. Bilirubin and liver enzymes returned to normal within three months of lapatinib discontinuation. Lapatinib should be included among the causes of drug-induced hepatitis