Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. As usually HCC diagnosis occurs in the advanced stage of the disease, available treatments such as resection, liver transplant, or local ablation are poorly if not at all effective. Also systemic chemotherapy has limited effectiveness. The only drug that can prolong patient survival is Sorafenib; unfortunately, however, the extent of increased survival is very modest being of few months. Together the above considerations clearly indicate that the development of novel therapeutic approaches for HCC are urgently necessary. The demethylation agent 5-Azacytidine (5-AZA) is considered to be of great potential therapeutic value. This is due to the emerging knowledge of the important role of epigenetic modulation in HCC development and to the fact that the use of demethylation agents gave encouraging results in cultured human HCC derived cells. Working with different cellular models of HCC (JHH6, HUH7, IHH) we got evidence of the potent anti-proliferative (cell counting, MTT, flow-cytometry, immuno-fluorescence) and anti-migratory (scratch assay, trans-well test, microscopic live observation of migration) effects of 5-AZA. Functional molecular analysis revealed that, upon 5-AZA treatment, a strong up-regulation of miRNA 139-5p occurs. This in turn causes the down regulation of the Rho-associated coiled-coil-containing protein kinase 2 (ROCK2), a known target of miRNA 139-5p. ROCK2 inhibition then results in the reduced stabilization of the gelatinase MMP2, paralleled by the reduction of MMP2 activity (as evaluated by zymography). In addition, ROCK2 down regulation leads to the decrease in the expression of other targets including Cyclin D1, E2F1 and Pin-1, all proliferative genes. Thus, the potent anti-migratory and anti-proliferative effects of 5-AZA may occur via the inhibition of pathways miRNA139-5p/ROCK2/MMP2 and miRNA139-5p/ROCK2/CyclinD1/E2F1/Pin-1 pathway, respectively. Finally, we have observed in a preliminary experiment in SCID xenograft mouse model of HCC that the intra-tumour injection of 5-AZA administered in two consecutive rounds, can significantly reduce tumour growth prolonging animal survival. The effectiveness of 5-AZA against HCC we observed can significantly contribute to bring 5-AZA closer to the clinical use for HCC.

Exploring drug molecular effects in cancer disease models / Tonon, Federica. - (2016 Apr 20).

Exploring drug molecular effects in cancer disease models

TONON, FEDERICA
2016-04-20

Abstract

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related death worldwide. As usually HCC diagnosis occurs in the advanced stage of the disease, available treatments such as resection, liver transplant, or local ablation are poorly if not at all effective. Also systemic chemotherapy has limited effectiveness. The only drug that can prolong patient survival is Sorafenib; unfortunately, however, the extent of increased survival is very modest being of few months. Together the above considerations clearly indicate that the development of novel therapeutic approaches for HCC are urgently necessary. The demethylation agent 5-Azacytidine (5-AZA) is considered to be of great potential therapeutic value. This is due to the emerging knowledge of the important role of epigenetic modulation in HCC development and to the fact that the use of demethylation agents gave encouraging results in cultured human HCC derived cells. Working with different cellular models of HCC (JHH6, HUH7, IHH) we got evidence of the potent anti-proliferative (cell counting, MTT, flow-cytometry, immuno-fluorescence) and anti-migratory (scratch assay, trans-well test, microscopic live observation of migration) effects of 5-AZA. Functional molecular analysis revealed that, upon 5-AZA treatment, a strong up-regulation of miRNA 139-5p occurs. This in turn causes the down regulation of the Rho-associated coiled-coil-containing protein kinase 2 (ROCK2), a known target of miRNA 139-5p. ROCK2 inhibition then results in the reduced stabilization of the gelatinase MMP2, paralleled by the reduction of MMP2 activity (as evaluated by zymography). In addition, ROCK2 down regulation leads to the decrease in the expression of other targets including Cyclin D1, E2F1 and Pin-1, all proliferative genes. Thus, the potent anti-migratory and anti-proliferative effects of 5-AZA may occur via the inhibition of pathways miRNA139-5p/ROCK2/MMP2 and miRNA139-5p/ROCK2/CyclinD1/E2F1/Pin-1 pathway, respectively. Finally, we have observed in a preliminary experiment in SCID xenograft mouse model of HCC that the intra-tumour injection of 5-AZA administered in two consecutive rounds, can significantly reduce tumour growth prolonging animal survival. The effectiveness of 5-AZA against HCC we observed can significantly contribute to bring 5-AZA closer to the clinical use for HCC.
20-apr-2016
GRASSI, Mario
28
2014/2015
Settore CHIM/06 - Chimica Organica
Università degli Studi di Trieste
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11368/2908483
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