Adlay Seed (Coix lacryma-jobi L. var. Ma-yuen Stapf.) Ethanolic Extract Fractions and Subfractions Induce Cell Cycle Arrest and Apoptosis in Human Breast and Cervical Cancer Cell Lines

Abstract

The antitumor effects of Coix lacryma-jobi L. var. ma-yuen Stapf. (adlay seed) ethanolic extract have been increasingly shown. This study aimed to investigate the beneficial effects of both the fractions and subfractions of adlay seed ethanolic extract on the human breast (MCF-7) and cervical (HeLa) cancer cell lines, as well as exploring their possible mechanisms of action. The ethanolic extracts were obtained from different parts of adlay seed, including AHE (adlay hull extract), ATE (adlay testa extract), ABE (adlay bran extract) and PAE (polished adlay extract). The results of a 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl- tetrazolium bromide (MTT) assay showed that AHE-Ea and ATE-Ea showed significant growth inhibitory effects in a dose-dependent manner. The results also showed that the AHE-Ea-K, AHE-Ea-L, ATE-Ea-E and ATE-Ea-F subfractions inhibited cell proliferation, induced cell cycle arrest in the G0/G1 phase and decreased CDK4/Cyclin D1 protein expression. Finally, the extract activated caspase-3 activity and PARP protein expression, which induced MCF-7 and HeLa cell apoptosis. We then used liquid chromatography–mass spectrometry (LC/MS) to identify the potential active components., Quercetin showed an anticancer capacity. In conclusion, the AHE-Ea-K, AHE-Ea-L, ATE-Ea-E and ATE-Ea-F subfractions showed antitumor effects through the inhibition of MCF-7 and HeLa cell line viability, as well as inducing apoptosis and cell cycle arrest.

Cardiovascular protection of YiyiFuzi powder and the potential mechanisms through modulating mitochondria-endoplasmic reticulum interactions

“…An in-vitro study demonstrated that CSO upregulates the PTEN protein while inhibiting the expression of p-AKT and p-PI3K proteins, thereby inducing apoptosis in human pancreatic cancer cells through the PTEN/PI3K/AKT signaling pathway ( Yang et al, 2022a ). Additionally, Coicis semen has been found to inhibit proliferation and promote the apoptosis of various cancer cell types, including cancer cells of the lung ( Chang et al, 2003 ), colon ( Lee et al, 2008 ), hepatic ( Qu et al, 2016 ), mouth ( Manosroi et al, 2019 ), breast ( Fang et al, 2020 ; Chiang et al, 2022 ), uterine ( Huang et al, 2021 ), cervical ( Chen et al, 2018 ; Guo et al, 2019 ; Chiang et al, 2022 ), and pancreatic ( Yang et al, 2022a ). While not directly related to cardiovascular health, this could contribute to overall health and wellbeing ( Zeng et al, 2022 ).…”

The Adipokine Visfatin Modulates Cancer Stem Cell Properties in Triple-Negative Breast Cancer

“…Tumor samples (about 0.1 g) were homogenized with lysis buffer containing a protease inhibitor (Roche, Basel, Switzerland) and a phosphatase inhibitor (Roche) using TissueLyser II (Qiagen, Chatsworth, CA, USA) and centrifuged at 12,000× g for 30 min at 4 • C. A BCA kit (T-Pro Biotechnology, New Taipei City, Taiwan) was used to measure the tissue and serum protein concentration. We used 10-15% SDS-polyacrylamide gel electrophoresis (PAGE) for protein separation and then transferred the samples onto Immobilon-P polyvinylidene fluoride (PVDF) membranes (0.22 µm) for 125 min at 95 V. Then, we used blocking buffer (5% BSA) for 1 h at room temperature and incubated the samples with primary antibodies against NAMPT (Proteintech, Rehovot, Israel), NANOG (Proteintech), OCT4 (Proteintech), SOX2 (Proteintech), SIRT1 (Cell signaling, Boston, MA, USA), VEGF (Santa Cruz Biotechnology, Santa Cruz, CA, USA), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Proteintech, Rehovot, Israel) overnight at 4 • C. After washing 3 times, the samples were incubated with anti-rabbit/mouse IgG coupled with alkaline phosphatase (1: 10,000) for 2 h. We used an ECL chemiluminescent kit to visualize the antibody-antigen interaction and detected the signal with an eBlot Touch Imager (eBlot Photoelectric Technology, Shanghai, China) [20]. The relative intensity was measured using ImageJ software (NIH, Bethesda, MD, USA).…”

Potential of the Novel Slot Blot Method with a PVDF Membrane for Protein Identification and Quantification in Kampo Medicines

“…and Mashinin (Cannabis Fructus). Some studies have analyzed lowmolecular compounds or polysaccharides that can be extracted into a low-polarity solvent in Yokuinin and investigated their effects in vitro and in vivo [63][64][65][66][67][68][69]. However, Li et al extracted the components in Yokuinin using high-polarity solvents, namely, 0.5 M sodium chloride aqueous, 70% ethanol aqueous, and 12.5 mM sodium borate buffer [18]; these four solvents contained albumin, globulin, prolamin, and glutelin, and their target was glutelin.…”