Background and aim: Cholangiocarcinoma (CC) is characterized by a poor prognosis. As curative medical regimen is currently unavailable for patient unsuitable for surgery, new drugs are urgently needed. Resveratrol (RES) has previously been reported to play a cytotoxic effect on CC cell cultures and the related increase of type 2 transglutaminase (TG2) involved in carcinogenesis and apoptosis. Present study was aimed at evaluating if TG2 inhibition could reduce the cytotoxic effect of RES on CC cell lines. Material and methods: CC cell lines SK-CHA-1 and MZ-CHA-1, grown on a three dimensional cell culture model, were treated for 72 hours with RES (64 microM, after a dose finding preliminary study) alone or combined with TG2 inhibitors (cystamine and two selectives ones, B003 and T101). The following points were investigated: cell viability (clonigenic test), cell morphology (light microscopy -LM, transmission electron microscopy -TEM and immunoistochemistry-IMC), Q-banding (karyotype analysis), and TG2 analysis (colorimetric method and Western Blotting). Results: RES treatment induced a significant inhibition of cell growth. The co-treatment RES/TG2 inhibitors prevented growth inhibition in both cell lines; the cell growth for SK-CHA-1 with RES 64 microM and the three different inhibitors (respectively cystamine, B003 and T101) increased of the 24%, 42% and 76% vs. percentage of colony growth from cells treated only with RES; for MZ-CHA-1 the cell growth increase was of 75%, 33% and 83% (cystamine, B003 and T101) vs. percentage of colony growth from cells treated only with RES. LM, TEM and IHC results demonstrated a partial protection with T101. The normalization of cell growth was associated to an inhibition of TG2 activity both in MZ-CHA-1 (85% with cystamine, 60% with B003 and 45% with T101 vs.100% controls) and SK-CHA-1 (95% with cystamine, 30% with B003 and 15% with T101 vs. 100% controls). Conclusions: Our data demonstrated that the cytotoxic effect of RES in SK-CHA-1 and MZ-CHA-1 is TG2 mediated.
Transglutaminase 2 (TG2) mediates the cytotoxic effect of resveratrol in cholangiocarcinoma (CC) cell lines / L.R. Roncoroni, E. L., B. P., L. Tacchini, L. V., B. F., D. Conte, L. Doneda. ((Intervento presentato al 22. convegno National Congress of Digestive Diseases tenutosi a Napoli nel 2016.
Transglutaminase 2 (TG2) mediates the cytotoxic effect of resveratrol in cholangiocarcinoma (CC) cell lines
L.R. Roncoroni;L. Tacchini;D. Conte;L. Doneda
2016
Abstract
Background and aim: Cholangiocarcinoma (CC) is characterized by a poor prognosis. As curative medical regimen is currently unavailable for patient unsuitable for surgery, new drugs are urgently needed. Resveratrol (RES) has previously been reported to play a cytotoxic effect on CC cell cultures and the related increase of type 2 transglutaminase (TG2) involved in carcinogenesis and apoptosis. Present study was aimed at evaluating if TG2 inhibition could reduce the cytotoxic effect of RES on CC cell lines. Material and methods: CC cell lines SK-CHA-1 and MZ-CHA-1, grown on a three dimensional cell culture model, were treated for 72 hours with RES (64 microM, after a dose finding preliminary study) alone or combined with TG2 inhibitors (cystamine and two selectives ones, B003 and T101). The following points were investigated: cell viability (clonigenic test), cell morphology (light microscopy -LM, transmission electron microscopy -TEM and immunoistochemistry-IMC), Q-banding (karyotype analysis), and TG2 analysis (colorimetric method and Western Blotting). Results: RES treatment induced a significant inhibition of cell growth. The co-treatment RES/TG2 inhibitors prevented growth inhibition in both cell lines; the cell growth for SK-CHA-1 with RES 64 microM and the three different inhibitors (respectively cystamine, B003 and T101) increased of the 24%, 42% and 76% vs. percentage of colony growth from cells treated only with RES; for MZ-CHA-1 the cell growth increase was of 75%, 33% and 83% (cystamine, B003 and T101) vs. percentage of colony growth from cells treated only with RES. LM, TEM and IHC results demonstrated a partial protection with T101. The normalization of cell growth was associated to an inhibition of TG2 activity both in MZ-CHA-1 (85% with cystamine, 60% with B003 and 45% with T101 vs.100% controls) and SK-CHA-1 (95% with cystamine, 30% with B003 and 15% with T101 vs. 100% controls). Conclusions: Our data demonstrated that the cytotoxic effect of RES in SK-CHA-1 and MZ-CHA-1 is TG2 mediated.
Pubblicazioni consigliate
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
