Targeting transforming growth factor (TGF)-betaRI inhibits activation of beta1 integrin and blocks vascular invasion in hepatocellular carcinoma.

UNLABELLED: Vascular invasion is one of the major negative prognostic factors in patients with hepatocellular carcinoma (HCC), leading to cancer recurrence. To invade, HCC cells must penetrate the vessel wall, consisting of endothelial cells and extracellular matrix components, including fibronectin and fibrinogen. Employing invasive and noninvasive HCC cells, we studied the mechanism underlying vascular invasion. We show that HCC cells invade blood vessels via alpha5beta1, that is equally expressed in invasive and noninvasive cells. However, in the former, the intracytoplasmic tail of beta1 integrin is constitutively phosphorylated at threonine 788-789 and the extracellular part is conformationally activated. In noninvasive cells, beta1 integrin is not activated. Transforming growth factor (TGF)-beta1 specifically phosphorylates beta1 integrin (threonine 788-789) via Smad-2 and Smad-3, causing a conformational change of the extracellular component with an inside-out mechanism. This leads noninvasive HCC cells to behave like invasive cells. A selective TGF-betaRI inhibitor inhibits phosphorylation of the beta1 integrin intracytoplasmic tail, and blocks invasion of HCC cells, both constitutively invasive and with acquired invasive properties. In human HCC tissues with microvascular invasion, phospho-beta1 integrin was detected as well as TGF-beta1, p-Smad-2, and E-cadherin.
CONCLUSION: TGF-beta1 promotes vascular invasion by activating beta1 integrin. This suggests a rationale for targeting TGF-betaRI in future clinical trials.