Anticancer property of Selaginella bryopteris (L.) Bak. against hepatocellular carcinoma in vitro and in vivo

Hepatocellular carcinoma (HCC) is the most common and leading cause of cancer-related mortality worldwide. It has limited therapeutic options and side effects caused by current treatment methods. Selaginella bryopteris is used traditionally in the treatment of several health-related problems but mostly remains to be scientific validation. This study investigated the effect of Selaginella bryopteris on a hepatic cancer cell line in vitro and in the NDEA-induced HCC rat model in vivo. The anticancer effect of S. bryopteris was evaluated in vitro using a human hepatocellular carcinoma HepG2 cell line. Gas chromatography-mass spectrometry (GC–MS) was carried out to analyze the phytoconstituents of S. bryopteris extract. Different techniques were used in this in vitro anticancer study to evaluate cell viability, alteration in nuclear morphology, mitochondrial membrane potential, and generation of reactive oxygen species (ROS). For an in vivo study, Sprague Dawley (SD) rats were induced with N- nitrosodiethylamine (200 mg/kg b.w. once, i.p.) and carbon tetrachloride (3 mL/kg/week b.w., s.c.) up to six weeks. The induced animals were treated with S. bryopteris extract (100 and 200 mg/kg) and silymarin (100 mg/kg) for 28 days. The level of liver function parameters and antioxidant enzymes were assessed. A reverse transcriptase-polymerase chain reaction was used to evaluate the gene expression patterns of p53, Bax, caspase-3, caspase-9, and Bcl-2 mRNAs. In a liver cancer cell line (HepG2), Selaginella bryopteris extract (SBE) induced apoptosis through altering mitochondrial membrane potential, nuclear condensation, and the production of reactive oxygen species. The elevated level of ALT, AST, GGT, and TBL in HCC-induced rats were declined significantly by SBE administration. SBE enhanced antioxidant activity (GST, Catalase, SOD, GPX), antioxidant molecule (GSH), and reduced the level of stress marker (MDA) in NDEA induced animals. The effects of SBE against HCC were also demonstrated by the up-regulated expression of pro-apoptotic genes (p53, caspase-3, caspase-9, Bax) and the down-regulated expression of anti-apoptotic (Bcl-2) gene by qRT-PCR analysis in liver tissue. Histological analysis revealed that the extract improved the liver architecture. The major bioactive compounds detected by GC–MS analysis were imidazole, amentoflavone, gallic acid, catechine, palmitic acid, lupeol, l-Fucitol, and myo-inositol. SBE was found to be safe in an acute toxicity test. As a