Role of H2S and TGF-β1 in the angiogenic process and in the development of Hepatocarcinoma

DEZA, Z; RIDRUEJO, E; ROMERO-CAÍMI, G; SAENZ, D; COLI, L; CHIAPPINI, F; CALVO, G; DI VENOSA, G; ALVAREZ, L

Congreso; Reunion Anual de Sociedades de Biociencias 2020 - SAIC-SAI-SAFIS; 2020

Sociedad Argentina de Investigación Clínica y otras (SAIC-SAI-SAFIS)

Hepatocarcinoma (HCC) represents 90% of primary liver tumours. Hexachlorobenzene (HCB) is a dioxin-type toxic and promoter of preneoplastic foci in rat liver. We have shown that it alters regulatory parameters of cell growth in vivo and in vitro. In the present work, our objective was to evaluate the mechanisms of action of HCB in the development of HCC. We will study the key molecules present in the tumor microenvironment that collaborate in the angiogenesis process and in the development of HCC. We used in vivo, nude mice treated with HCB i.p. (3mg/kg) inoculated with HepG2. We evaluate: a) in vivo, PCNA (Western Blot (W));b) morphology(IH);c) tumours and d) angiogenesis (vessels/skin). In vitro, HepG2:a) PCNA (W);b) cell cycle (flow cytometry); c) p21,p27,TGF-B1(W);d) cD1 (W) and e) H2S. In Ea-hy926: a) PCNA (W), b) tubulogenesis (length of tubules); d) H2S and e) role of TGF-β1 in the effect of HCB(5μM) on PCNA (W) using specific inhibitor (SB431542); f) effect of the conditioned medium (CM) of HepG2 treated with HCB(5μM) on Cell Migration (Cmi). In vivo it increased in the HCB group: PCNA (30%,p<0.01); TGF-β1 (29%,p<0.01); mitotic nuclei; tumours, size 8 mmx9 mm/day 30 and vascularization, (35%,p<0.01). In HepG2, increase: PCNA (22, 31%, p<0.01) (0.5 and 5 μM HCB); S phase (21.26%,p<0.01) (0.5 and 5 μM HCB); TGF-β1 (20, 28, 40%,p<0.01) (HCB 0.05, 0.5 and 5μM) and cD1 (18, 25, 32%,p <0.01) (HCB 0.05 ; 0.5 and 5 μM). They decreased: p21 (14, 21, 28%,p<0.05); p27 (19, 27, 31%,p<0.01) (HCB 0.05, 0.5 and 5 μM) and H2S (20, 27%,p<0.05) (HCB 0.5 and 5 μM) . In Ea-hy926 increased: a) PCNA (40%, p<0.01) and tubule length (56%,p<0.01) and decreased: H2S (21%,p<0.05). PCNA did not vary when treating with SB431542/HCB (5 μM). The CM of HepG2/HCB(5μM) increased (CMi) (32%, p<0.01). We conclude that in the in vivo model of HCC, HCB deregulates cell growth and favours the development of neo-angiogenesis, H2S and TGF-β1 molecules would be involved in both processes.