Monotropein Induced Apoptosis and Suppressed Cell Cycle Progression in Colorectal Cancer Cells.

OBJECTIVE: To determine whether monotropein has an anticancer effect and explore its potential mechanisms against colorectal cancer (CRC) through network pharmacology and molecular docking combined with experimental verification. METHODS: Network pharmacology and molecular docking were used to predict potential targets of monotropein against CRC. Cell counting kit assay, plate monoclonal assay and microscopic observation were used to investigate the antiproliferative effects of monotropein on CRC cells HCT116, HT29 and LoVo. Flow cytometry and scratch assay were used to analyze apoptosis and cell cycle, as well as cell migration, respectively in HCT116, HT29, and LoVo cells. Western blotting was used to detect the expression of proteins related to apoptosis, cell cycle, and cell migration, and the expression of proteins key to the Akt pathway. RESULTS: The Gene Ontology and Reactome enrichment analyses indicated that the anticancer potential of monotropein against CRC might be involved in multiple cancer-related signaling pathways. Among these pathways, RAC-beta serine/threonine-protein kinase (Akt1, Akt2), cyclin-dependent kinase 6 (CDK6), matrix metalloproteinase-9 (MMP9), epidermal growth factor receptor (EGFR), cell division control protein 42 homolog (CDC42) were shown as the potential anticancer targets of monotropein against CRC. Molecular docking suggested that monotropein may interact with the 6 targets (Akt1, Akt2, CDK6, MMP9, EGFR, CDC42). Subsequently, cell activity of HCT116, HT29 and LoVo cell lines were significantly suppressed by monotropein (P<0.05). Furthermore, our research revealed that monotropein induced cell apoptosis by inhibiting Bcl-2 and increasing Bax, induced G1-S cycle arrest in colorectal cancer by decreasing the expressions of CyclinD1, CDK4 and CDK6, inhibited cell migration by suppressing the expressions of CDC42 and MMP9 (P<0.05), and might play an anticancer role through Akt signaling pathway. CONCLUSION: Monotropein exerts its antitumor effects primarily by arresting the cell cycle, causing cell apoptosis, and inhibiting cell migration. This indicates a high potential for developing novel medication for treating CRC.

Determination of Maintenance Energy Requirements for Fattening Castrated Korean Black Goats (Capra hircus coreanae).

Twelve adult (10 months old) castrated Korean black goats, with an average initial body weight of 24.98 ± 3.7 kg, were used in this experiment to determine their maintenance energy requirements. Dry matter intakes (g/d, p = 0.945) were not affected by energy levels, but metabolic energy intake (kcal/d, p < 0.002) and average daily gain (g/d, p < 0.001) were significantly increased at higher energy levels. Nutrient digestibility was similar in the treatments, but crude fat digestibility increased with the addition of protective fat powder (p = 0.001). The energy required for fattening the castrated Korean black goats was estimated using the correlation between metabolic energy intake per dietary body weight and average daily gain per dietary body weight. The Y-axis intercept value was calculated to be 108.76 kcal/kg BW0.75 (p < 0.05, r2 = 0.6036), which was the metabolic energy requirement for maintaining the lives of the fattening Korean black goats. The estimated energy requirements of the black goat can improve specification techniques, such as the energy level and the amount of feed supply required for domestic black goats.