A Combined Treatment with Berberine and Andrographis Exhibits Enhanced Anti-Cancer Activity through Suppression of DNA Replication in Colorectal Cancer

Abstract

The high morbidity and mortality associated with colorectal cancer (CRC) are largely due to the invariable development of chemoresistance to classic chemotherapies, as well as intolerance to their significant toxicity. Many pharmaceutical formulations screened from natural plant extracts offer safe, inexpensive, and multi-target therapeutic options. In this study, we demonstrated that Berberis vulgaris L. (Berberine) and Andrographis paniculata (Burm. f.) Nees (Andrographis) extracts exerted their synergistic amplified anti-cancer effects by jointly inhibiting cell viability, suppressing colony formation, and inducing cell cycle arrest. Consistent with our in-vitro findings, the amplified synergistic anti-cancer effects were also observed in subcutaneous xenograft preclinical animal models, as well as patient-derived primary tumor organoids. To explore the molecular mechanisms underlying the amplified synergistic anti-cancer effects, RNA sequencing was performed to identify candidate pathways and genes. A transcriptome analysis revealed that DNA-replication-related genes, including FEN1, MCM7, PRIM1, MCM5, POLA1, MCM4, and PCNA, may be responsible for the enhanced anticancer effects of these two natural extracts. Taken together, our data revealed the powerful enhanced synergistic anti-CRC effects of berberine and Andrographis and provide evidence for the combinational targeting of DNA-replication-related genes as a promising new strategy for the therapeutic option in the management of CRC patients.

Curcumin and Andrographis Exhibit Anti-Tumor Effects in Colorectal Cancer via Activation of Ferroptosis and Dual Suppression of Glutathione Peroxidase-4 and Ferroptosis Suppressor Protein-1

“…Colony formation assays were performed as previously reported [ 23 , 40 ]. Five hundred cancer cells were seeded in 6-well plates.…”

“…For these experiments, 2.5 × 10 5 cells from SW480 and HCT116 lines were treated with each compound for 16 h before total RNA extraction using the MiRNeasy Mini kit (Qiagen, Hilden, Germany). The details for RNA-sequencing library preparation were previously reported [ 12 , 23 , 40 ]. Briefly, the library was constructed using the TruSeq RNA Sample Prep Kit (Illumina, San Diego, CA) with 1 μg of purified RNA input.…”

“…In this context, curcumin and other naturally-occurring botanicals have also been shown to be ferroptosis inducers in cancer cells [ 5 , 16 , 20 , 21 , 22 ]. We previously reported that andrographis, an active compound derived from the Chinese herbal medicine andrographis paniculate, exhibited anti-tumor activity in gastric and CRC via the activation of the ferroptosis pathway [ 21 , 22 , 23 , 24 , 25 ]. Based on these studies, data suggest that curcumin and andrographis exhibit an anti-tumor effect in cancer by activating the ferroptosis pathway.…”

Berberine Overcomes Gemcitabine-Associated Chemoresistance through Regulation of Rap1/PI3K-Akt Signaling in Pancreatic Ductal Adenocarcinoma

“…Cell cycle and apoptosis assays were performed using the Muse Cell Analyzer (EMD Millipore Corp, Hayward, CA, USA) according to the manufacturer’s instructions [ 25 , 26 , 33 , 84 , 85 , 86 , 87 , 88 , 89 , 90 , 91 ]. For the cell cycle assays, cells were seeded in 6-well plates and treated with BBR, Gem, and their combination for 48 h. Thereafter, cells were rinsed with ice-cold phosphate-buffered saline (PBS), fixed with 200 μL of ice-cold 70% ethanol and incubated at −20 °C for more than 3 h. Two hundred microliters of Muse TM Cell Cycle Reagent (EMD Millipore Corp) were used in staining cells.…”

Pharmacokinetic of berberine, the main constituent of Berberis vulgaris L.: A comprehensive review

“…A major constituent of B. vulgaris is berberine which is an isoquinoline alkaloid (a 5,6‐dihydrodibenzo[a,g]quinolizinium) that has various uses in Ayurveda and Traditional Chinese Medicine (McCubrey et al, 2022; Sadeghnia, Kolangikhah, Asadpour, Forouzanfar, & Hosseinzadeh, 2017; Sadeghnia, Taji, Forouzanfar, & Hosseinzadeh, 2017; Xia et al, 2022). Berberine is also present in many other plants including further species of Berberis ( B. aristata L., B. croatica L. , B. aquifolium L.)., Coptis ( Coptis chinensis L., Coptis japonica L.), and Hydrastis ( Hydrastis Canadensis L.) (Imenshahidi & Hosseinzadeh, 2019) and has many pharmacologic properties (Jang, Piao, Kim, Kwon, & Park, 2008), such as antioxidant (Shou et al, 2022), anti‐microbial (Tian et al, 2022), antiinflammatory (Deng et al, 2022), anticancer (Zhao, Roy, Wang, & Goel, 2022), anti‐diabetes (Zhang et al, 2022), anti‐dyslipidemia (Zhao et al, 2021), anti‐obesity (Bhattacharjee, Nath, & Choudhury, 2021), and anti‐metabolic syndrome (Tabeshpour, Imenshahidi, & Hosseinzadeh, 2017). A relatively large number of clinical studies have shown that berberine can display its therapeutic activities in the treatment of different diseases including diabetes, hyperlipidemia, cancer, hypertension, stroke, metabolic syndrome, polycystic ovarian syndrome, and liver disease in human subjects.…”