Hypericin Has the Potential to be used as Therapeutic Compound in Liver Cancer

Primary liver cancer, also known as hepatocellular carcinoma (HCC), is one of the most lethal cancers having worldwide prevalence. Although most HCC cases are reported in the developing countries of Asia and Africa, there has been an alarming increase in HCC cases in Western Europe as well as United States. Chronic liver diseases, viral hepatitis, alcoholism as well as dietary carcinogens, such as aflatoxins and nitrosamines, contribute to HCC and most often, develops and progresses in a milieu of oxidative stress and inflammation. Phytochemicals, such as dietary polyphenols endowed with potent antioxidant as well as anti-inflammatory properties, provide a suitable alternative in affording alleviation of HCC. Hypericin, the principal of Hypericum Perfuratum, has long been used to cure several chronic ailments, such as neoplastic and neurodegenerative diseases.Materials & Methods: Cells were incubated in cell culture incubator with these conditions: ۳۷°C with ۵% CO۲; and changing media every ۳ or ۴ days. When confluency of these cells reached into ۸۰-۹۰%, cells were detached by trypsin. After neutralization and centrifugation, cells were counted and utilized for subsequent tests to perform real time PCR, total RNA was extracted from each sample treated with appropriate dose of hypericin by TRIzol reagent (Invitrogen). To further investigate different expression of p۵۳ protein in various samples, ICC test utilized. To perform ICC, cells were seeded in ۱۲ well plates and treated with appropriate dose of Hypericin. These cells were then fixed by ۴% of paraformaldehyde for ۱۰min at room temperature. After that cells were incubated with antibody against p۵۳ protein for ۱۶ hours at ۴°C.Findings: MTT results show that, Hypericin has the ability to induce ۵۰% of cellular death (LD۵۰) at the concentration of ۲ (μg/ml) in ۲۴ hours and ۱(μg/ml) in ۴۸ hours. Annexin V/PI test demonstrated that, cellular death induced by Hypericin was kind of apoptosis. Treatment of Huh۷ cells lines with LD۵۰ dose of Hypericin for ۲۴ hours, induced ۵۳% apoptosis. To further confirm that Hypericin induces apoptosis in Huh۷ cells, mRNA expression level of bax, bcl۲ and p۵۳ were investigated. Post treatment by LD۵۰ dose of Hypericin for ۲۴ hours, bcl۲ mRNA expression level decreased and p۵۳ and bax mRNA expression level increased significantly. Cells which treated by LD۵۰ dose of Hypericin, expressed p۵۳ protein much more than untreated cells.Conclusion: Results of the present study show that on HUH۷ cell line, the LD۵۰ of hypericin was lower than cisplatin and the rate of apoptosis in treatment of cells with hypericin was lower than cisplatin. Further studies are needed to evaluate the chemo preventive potential of the hypericin when used alone or in combination with cisplatin to mitigate the toxic side effects of the latter

کلیدواژه ها:

Chemoprevention ، Hypericin ، Huh۷ ، liver cancer