Abstract
ANTICANCER EFFECT OF BENZYL ISOTHIOCYANATE IN COMBINATION WITH LOW DOSE OF GAMMA RADIATION AGAINST HEPATOCELLULAR CARCINOMA PROLIFERATION INDUCED BY DIETHYLNITROSAMINE

Iman Hesham El-Khashab*

ABSTRACT

Objective: Benzyl isothiocyanate (BITC) or low dose of radiation are considered as cancer therapeutic agents. The aim of this study was to evaluate the anticancer activity and the antioxidant capacity of Benzyl isothiocyanates combined with the low dose of γ-irradiation against liver hepatotoxicity and hepatocellular carcinoma induced in rats by Diethylnitrosamine (DEN). Also, to evaluate apoptosis by estimation of protein expression of caspase-3, Bax and Bcl-2 in hepatocellular carcinoma. Methods: Fifty male Wistar rats were divided into five groups. Group (1): Negative control group (C), Group (2) DEN group: rats were intraperitoneally injected with a single dose of Diethylnitrosamine (200 mg/kg) Group (3) DEN+ BITC: rats were injected intraperitonealy with a single dose of DEN 200 mg/kg and orally administrated with BITC at a dose of 10 mg/kg three times per week for ten weeks. Group(4) DEN+R: rats were injected intraperitonealy with a single dose of DEN 200 mg/kg and irradiated with low dose of γ-irradiation (0.25gy) applied on the 1st and the 35th day. Group(5) DEN+ BITC+ R: rats were injected intraperitonealy with a single dose of DEN 200 mg/kg, orally administrated with BITC at a dose of 10 mg/kg three times per week for ten weeks and irradiated with low dose of γ-irradiation (0.25gy) applied on the 1st and the 35th day. Results: Histopathological study illustrated Hepatocellular carcinoma (HCC) features induced by DEN seen as an aggregation of tumor cell nodules surrounded by fibrotic tissue; formation of new blood vessels between tumor cells “angiogenesis”; loss of hepatic architecture forming nests of pleomorphic tumor hepatocytes; hydropic degeneration of hepatocytes with necrotic hepatocytes, and also dilatation in blood sinusoids. In addition, abundant collagen fiber deposition around central veins and portal tracts in HCC with Masson's trichrome staining were also noticed. Treatment with Benzyl isothiocyanates and/or low dose of radiation showed high amelioration in cancer cells progression and limitation of angiogenesis and marked recovery in hepatic architecture. Also, marked attenuation in collagen fiber deposition around central veins and portal tracts in treated groups were seen. Immunohistochemical expression in rats treated with Benzyl isothiocyanates and/or low dose of radiation showed regulation in the antiapoptotic protein Bcl2 and in the two proapoptotic proteins Bax and Caspase-3. Conclusion: Results indicated that treatment with Benzyl isothiocyanate combined with low dose of γ-irradiation has an effective therapeutic anticancer agent by inducing anti-angiogenesis and apoptosis in hepatocellular carcinoma. This effect is, in part, mediated through the mitochondrial pathway, which involves upregulation of the Bax/Bcl‑2 ratio and activation of caspase‑3 leading to marked apoptosis.

Keywords: Diethylnitrosamine, Benzyl isothiocyanate, Hepatocellular carcinoma, Apoptosis.


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