Background: Colorectal cancer (CRC) is the third leading cause of cancer associated mortality worldwide. Current available chemotherapeutics against CRC have various disadvantages. Medicinal plants are source of many biologically active compound(s) which have potential for development as therapeutic agents against different types of cancer. The aim of the present study was to screen for effective novel therapeutic agents against CRC from natural medicinal plants and herbs. Method:20 different plants from various species were collected randomly from various areas of Egypt. The cytotoxic and genotoxic effects against HCT-116 colon cancer cells was analyzed. Cytotoxicity was determined by MTT assay and comet assay was used to assess DNA damage. Gene expression was analyzed by qPCR for P53, Bcl2 and Bax. Results:The results revealed that the methanolic extracts of Ononis vaginalis and Quercus robur L. showed significant cytotoxic effect against HCT116 cells in vitro in monolayer and multicellular cancer spheroids(MCS). Further investigation of methanolic extracts treated HCT116 cells revealed marked fold increase in the expression of p53, Bax, whereas Bcl2 expres‌sion was significantly decreased. Moreover, the methanolic extract of O.vaginalis induced more DNA damage to colon cancer cells(HCT-116), the results was confirmed by the visual score which indicated that most of the effect was gradual increase in the length of the comet tail in parallel with a decrease in nuclear DNA content. This variation in DNA damage accounted for the observed genotoxic effect of O.vaginalis on HCT-116 cells in-vitro. Also, the methanolic extract of O.vaginalis significantly inhibited tumor growth in Syngeneic tumor model in C26 injected mice compared with the vehicle control group. Conclusion: Collectively, the findings of the present study indicated that methanolic extract of O.vaginalis was able to decrease proliferation and promote apoptosis in colon cancer cells by inducing the p53/Bax mediated signaling pathway.Therefore, It may be used as a novel source of new antitumor agent in the treatment of CRC.

The present study aimed to evaluate the chemopreventive potential of Pinus roxburghii branch (P. roxburghii) and Nauplius graveolens (N. graveolens) extracts against human colorectal cancer (CRC) induced by C26 murine cells in a BALB/c mouse model. Real-time qRT-PCR was used to evaluate the apoptotic pathway by measuring the relative mRNA expression levels of the Bcl-2, Bax, Cas3, NF-κB, and PI3k genes. At the termination of the 30-day period, blood samples were collected to assay the biomarkers. The results showed a significant increase (p < 0.05) in the levels of TGF-β, CEA, CA19-9, malondialdehyde, ALT, AST, ALP, urea, and creatinine in the positive control compared to the negative control group. In addition, the glutathione reductase activity and total antioxidant activity were reduced in the positive control compared to the negative control. The biomarkers mentioned above were restored to almost normal levels after administering a safe dose (1/10) of a lethal dose of P. roxburghii and N. graveolens extracts. Administration of one-tenth of the LD₅₀ of P. roxburghii and N. graveolens extracts caused a significant upregulation of the expression of Bax and Cas-3 and downregulation of the Bcl-2, NF-ĸB, and PI3k genes vs. the GAPDH gene as a housekeeping gene compared to the control group. Furthermore, the Bax/Bcl-2 ratio increased upon treatment. After administration of P. roxburghii and N. graveolens at a safe dose (1/10) of a lethal dose, the results showed improvement in both body weight gain and a significant decrease (p < 0.05) in tumor volume. Histopathological changes supported these improvements. Conclusively, the research outputs show that P. roxburghii and N. graveolens extracts can be utilized as potential chemopreventive agents for CRC treatment by stimulating cancer cell apoptosis and suppressing CRC survival and proliferation.

The use of phytochemicals is gaining increasing attention for treating cancer morbidity with minimal burden side effects. This study evaluated the cytotoxicity and antimicrobial activities of Pinus roxburghii branch (P. roxburghii) and Nauplius graveolens (N. graveolens) extracts in vitro. Cell viability was estimated using MTT assay. DNA fragmentation was determined to detect apoptotic pathway initiation. Mechanistically, the apoptotic pathway was tracked by estimating the relative mRNA expression levels of the Bcl-2, Bax, Cas3, NF-κB, and PI3k genes by qRT-PCR. P. roxburghii exhibited moderate antioxidant activity, while N. graveolens possessed highly significant (p < 0.05) scavenging activity against DPPH and ABTS assays. HPLC analysis demonstrated that catechin and chlorogenic acid were the predominant polyphenolic compounds in P. roxburghii and N. graveolens, respectively. The P. roxburghii and N. graveolens extracts inhibited the viability of HCT-116 cells with IC₅₀ values of 30.6 µg mL⁻¹ and 26.5 µg mL⁻¹, respectively. DNA fragmentation analysis showed that the proposed extracts induced apoptosis in HCT-116 cells. Moreover, the IC₅₀ doses of the selected extracts significantly (p < 0.05) upregulated Bax and cleaved Cas-3, and downregulated Bcl-2, NF-κB, and PI3k genes versus the GAPDH gene as a housekeeping gene in comparison to the control group. The Bax/Bcl-2 ratio was raised upon treatment. The mentioned extracts exhibited antimicrobial action against all tested bacteria and fungi. The highest antibacterial effect was recorded against E. coli, with inhibition zones of 12.0 and 11.2 mm for P. roxburghii and N. graveolens, respectively. On the other hand, the highest antifungal action was registered for Penicillium verrucosum and A. niger, with inhibition zones of 9.8 and 9.2 mm for the tested extracts, respectively. In conclusion, the outcomes of this study indicate that P. roxburghii and N. graveolens extracts could potentially be used as anticancer, antibacterial, and antifungal agents.

The current study examined the anticancer potential of 80 synthetic organic compounds at 50µM using in vitro two-dimensional (2D) and three-dimensional (3D) spheroids model breast adenocarcinoma (MCF7) models. The most promising six cytotoxic compounds on the 3D spheroids model (> 70% growth cytotoxicity) were counter-screened for their safety on normal cells (RPE1 cell line) grown as 3D spheroids. In parallel, the six compounds were tested for their pro-apoptotic activity using the M30 Cyto death ELISA assay. Based on the activity on MCF7 3D spheroids model, half maximum inhibitory concentration (IC ₅₀ < 26µM), the safety on normal cells (< 35% cytotoxicity) and the pro-apoptotic activity on cancer cells (> fourfold increase of negative control), three compounds were considered the most promising and were selected for further investigation, named: F03, F06 and C02. These three candidates underwent further gene expression profiling of apoptosis and inflammatory related genes by quantitative real-time polymerase chain reaction (qRT-PCR). The gene expression results for the three compounds indicated favorable pro-apoptotic activity while unfavorable pro-inflammatory induction. Statistical analysis of the Xlog P values of the 3D spheroids model hits compared to that of the 80-compounds library showed a significant tendency for higher XlogP values in the 3D spheroids model hits sub-group, most probably due to the higher penetrability of lipophilic compounds into the tumor mass. The presented work can be considered a pilot model for an approach in anticancer drug discovery.

Background: Alzheimer's disease is a progressive brain disorder. Green tea (GT) and vitamin C are known for their antioxidant and anti-inflammatory properties. This study aims to investigate the potential oxidative stress, cell-based anti-inflammatory and genotoxic Profiles for synergistic effect of green tea and vitamin c against to Alzheimer's disease. Methods: In vitro assessments included their several antioxidant parameters and ability to inhibit inflammatory markers such as nitric oxide (NO), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and matrix metallopeptidase 1(MPP1) in lipopolysaccharide-activated macrophages. In vivo approach evaluated genotoxicity and their potential neurotherapy against Alzheimer's disease using a rat model. Behavioral, biochemical, and histological evaluations were conducted. Results: Green tea methanolic extract (GTME) and vitamin C showed enhanced antioxidant and anti-inflammatory modulator activities. The neurotoxicity rat’s model exhibited cognitive decline and brain injury in contrast to the control group. While treated rats with GTME and vitamin C exhibited improved cognitive function, behavior and reduced AChE levels the combination exerted a significantly more potent effect. Histological analysis confirmed improved brain tissue structure in rats receiving the combined treatment. Crucially, neither compound exhibited genotoxic properties. Conclusion: The combination of GTME and vitamin C demonstrated a promising therapeutic potential for neurodegenerative diseases due to their synergistic effects.