NF-kappaB is Not Directly Responsible for Photoresistance Induced by Fractionated Light Delivery in HT-29 Colon Adenocarcinoma Cells.

Abstract Our recent study follows up an earlier one which demonstrated hypericin-mediated photocytotoxic effects on HT-29 adenocarcinoma cells by light fractionation with a longer dark pause between two unequal light doses (Sackova, A. [2005] Photochem. Photobiol.81, 1411-1416). Here, we present closer study on events invoked by sublethal light dose (1 J cm(-2)) during the period of 6 h that is sufficient to invoke resistance to second lethal dose (11 J cm(-2)). First, we proved that the dark pause of 6 h, but not 1 h, resulted in better cell survival with suppressed phosphatidylserine externalization, decreased reactive oxygen species production and hypericin content as well as altered expression of HSP70, GRP94, clusterin, nuclear factor (NF)-kappaB, IkappaB-alpha or Mcl-1. NF-kappaB activity assay confirmed activation of this early-response pathway. However, inhibition of IkappaB (IKK) kinase by parthenolide by stopping NF-kappaB release from the complex with IkappaB did not prevent onset of resistance, but it invoked some resistance even in groups with shorter, 1 h dark pause. Therefore, we predict involvement of another signaling pathway, located upstream from NF-kappaB, responsible for onset of resistance to photodynamic therapy with hypericin in colon adenocarcinoma cells HT-29.