The phenolic and flavonoid content of the Stevia rebaudiana hydromethanolic extract (SRHME) was examined, and phytochemical identification using GC-mass spectrometry was achieved. Also, the antioxidant, antihemolytic, and antilipid peroxidation capabilities of the extract were assessed. The extract’s potential to induce apoptosis, inhibit cell proliferation, and reduce metastasis in SKVO3 cells was also checked. The findings of the GC-MS chromatogram demonstrated the existence of bioactive antioxidants and anticancer components in SRHME. Moreover, the extract demonstrated protection against cellular oxidative damage in human erythrocytes by preventing lipid peroxidation and hemolysis. Besides, SRHME demonstrated a selective cytotoxic effect with a strong IC50 value (17.5 µg/ml) on SKVO3 cells without any harmful effects on normal WI-38 cells using the MTT and LDH tests. Additionally, according to assays for wound healing and transwell chamber, the studied extract suppressed the ability of SKOV3 cells from migrating and invading, respectively. Also, the extract-treated SKVO3 cells showed rise in the percentage of apoptotic cells with a prominent comet nucleus, according to apoptotic assays in comparison to untreated cells. Furthermore, a flow cytometry analysis of SRHME-treated SKVO3 cells showed a halt in the S phase and an increase in sub-G1 apoptotic cells (25.44%). Also, the tested extract significantly decreased the levels of ROS in the treated cells, indicating that ROS was involved in the production of SKVO3 apoptosis. Lastly, SRHME strongly impacted the expression levels of proteins related to apoptosis, S-phase cell cycle arrest, and antimetastatic capacity in the treated SKVO3 cells.

Human interferon (IFN) is a type of cytokine that regulates the immune system’s response to viral and bacterial infections. Recombinant IFN-α has been approved for use in the treatment of a variety of viral infections as well as an anticancer medication for various forms of leukemia. The objective of the current study is to produce a functionally active recombinant human IFN-α2a from transgenic Raphanus sativus L. plants. Therefore, a binary plant expression construct containing the IFN-α2a gene coding sequence, under the regulation of the cauliflower mosaic virus 35SS promoter, was established. Agrobacterium-mediated floral dip transformation was used to introduce the IFN-α2a expression cassette into the nuclear genome of red and white rooted Raphanus sativus L. plants. From each genotype, three independent transgenic lines were established. The anticancer and antiviral activities of the partially purified recombinant IFN-α2a proteins were examined. The isolated IFN-α2a has been demonstrated to inhibit the spread of the Vesicular Stomatitis Virus (VSV). In addition, cytotoxicity and cell apoptosis assays against Hep-G2 cells (Human Hepatocellular Carcinoma) show the efficacy of the generated IFN-α2a as an anticancer agent. In comparison to bacterial, yeast, and animal cell culture systems, the overall observed results demonstrated the efficacy of using Raphanus sativus L. plants as a safe, cost-effective, and easy-to-use expression system for generating active human IFN-α2a.