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Anti-Blood Cancer Activities of Bioactive Compounds from Rice (Oryza sativa var. Koshihikari) and an Invasive Weed (Andropogon virginicus)

Anh La Hoang 広島大学

2022.09.20

概要

1. Background
Blood cancer (1.2 million cases per year) has been a serious human disease during a long history with a high mortality rate (58%). Therefore, this study was conducted to investigate promising candidates from plants for inhibiting blood cancer. Among plant sources, Adropogon virginicus is an invasive weed that distributes globally, threatening agriculture and economics in many countries. Thus, research on pharmaceutical properties may lead to further strategies to take advantage of this serious invasive weed that was once thought to be of no use. On the other hand, rice (Oryza sativa) is an important food crop in the world, which has been recently being received increasing attention for the research on medicinal purposes. In rice, momilactones are high valuable diterpene lactones exhibiting multiple biological benefits. Based on these reasons, we aimed (1) to explore potential phytochemicals and pharmaceutical properties of the invasive weed A. virginicus with focus on antioxidant, anti-skin aging, anti-diabetic, and anti-blood cancer properties and (2) to determine the potentials of momilactones isolated from rice (O. sativa var. Koshihikari) for preventing blood cancer.

2. Structure of dissertation
Chapter 1. General introduction
Chapter 2. Pharmaceutical properties of an invasive weed Andropogon virginicus
Chapter 3. Cytotoxic activities of momilactones against leukemia, lymphoma, and multiple myeloma cell lines
Chapter 4. Cytotoxic mechanism of momilactones against acute promyelocytic leukemia and multiple myeloma cell lines
Chapter 5. Effects of in vitro digestion on anti-blood cancer properties of momilactones Chapter 6. General discussion

3. Materials and methods
Materials
A. virginicus’ aerial plant parts were extracted by Soxhlet extraction method, followed by liquid-liquid phase extraction to obtain total crude (T-Anvi), hexane (H-Anvi), ethyl acetate, (E-Anvi), butanol (B-Anvi), and water (H-Anvi) extracts.

Momilactones were isolated and purified from the laboratory of Plant Physiology and Biochemistry, Hiroshima University, Japan (Quan et al., 2019b).

Biological activity
Antioxidant activities of samples were determined via antiradical (DPPH and ABTS), reducing power, β-carotene bleaching assays (Quan et al., 2019).

Enzymatic assays including tyrosinase and α-amylase inhibitory effects of A. virginicus’ extracts were evaluated following the method presented by Quan et al. (2019).

Anticancer assays consisting of MTT, annexin V apoptosis, cell cycle, and western blotting were conducted to screen the anticancer properties of momilactones. The corresponding cell lines for blood cancer were included leukemia (HL60, Meg-01, and K562), lymphoma (Mino), and myeloma (KMS-11 and U266). Besides, a non-cancerous cell line namely MeT-5A was used in this study.

In vitro digestion model
The effects of in vitro digestion on the bioaccessibility and cytotoxic activities of momilactones against blood cancer cell lines were investigated following the method described by Un et al. (2022).

Phytochemical analysis
The chemical analyses were conducted applying spectrophotometer, gas chromatography-mass spectrometry (GC-MS), and liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) methods.

4. Results and discussion
In Chapter 2, we indicate that the invasive weed A. virginicus may be a promising source of antioxidants, anti-⍺-amylase, and anti-tyrosinase abilities, and cytotoxicity against chronic myeloid leukemia cell lines. Besides, A. virginicus aerial parts are rich in flavonoids, palmitic acid, phytol, and γ-sitosterol, which may play a vital role in the biological activities of the respective extracts.

From Chapter 3, we highlight that MB and MAB can inhibit various blood cancer cell lines including leukemia, lymphoma, and multiple myeloma by inducing cell apoptosis. Notably, the cytotoxicity of MB and MAB is close to the well-known drugs of Doxorubicin in inhibiting Meg-01 and K562. Meanwhile these compounds are stronger than the widely applied medicines of Doxorubicin and Ibrutinib in suppressing HL-60 and U266, respectively. Whereas these compounds display an insignificant effect on non-cancerous cells.

In Chapter 4, MB and MAB at 5 µM promote acute promyelocytic leukemia (HL-60) and multiple myeloma (U266) cell apoptosis by activating the phosphorylation of p-38 in the mitogen-activated protein kinase (MAPK) pathway and regulating the relevant proteins (BCL-2 and caspase-3) in the mitochondrial pathway. Besides, these compounds may induce G2 phase arrest in HL-60 cell cycle through the activation of p-38 and disruption of CDK1 and cyclin B1 complex. Meanwhile MB and MAB show insignificant effects on cell apoptosis and cell cycle of non-cancerous cell line MeT-5A.

For Chapter 5, the finding shows that the bioaccessibility and cytotoxicity of momilactones significantly reduce through the digestive stages including oral, gastric, and intestinal phases.

A general discussion is presented in Chapter 6. Particularly, this is the first study to highlight that A. virginicus may be a promising source of antioxidant, anti-diabetic, anti-skin aging, and anti-blood cancer agents. In addition, momilactones isolated from rice (O. sativa var. Koshihikari) husks can inhibit blood cancer cell lines by regulating the proteins related to apoptosis and cell cycle pathways. However, cytotoxic activities of momilactones may be significantly decreased during digestion. Therefore, forthcoming studies should be conducted to improve the bioaccessibility of momilactones, followed by in vivo and clinical tests to confirm their capacity to develop novel anti-blood cancer drugs. In another aspect, the combination of momilactones from rice and isolated components from A. virginicus may be a potential approach for developing blood cancer treatments, especially for patients complicated with other health problems such as oxidative stress and chronic disorders.

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