Abstract
Shiitake (Lentinula edodes) and reishi mushrooms (Ganoderma sichuanense) have been noticed for desirable chemoprevention, such as improving immunity, overcoming anticancer drug resistance, etc. Particularly, beta-glucans have been mostly mentioned as the active component of the mushrooms. However, the right preparation and effective usage of the mushrooms for cancers are unclear, yet. Thus, we screened various extractions of the two mushrooms and combinations of the extracts, using a high throughput screening of 3D cell culture system on 384-pillar plates with various cancer cells and Cell Titer-Glo 3D cell viability assay kit (Promega). We also quantified the beta-glucans in all extractions with the Megazyme Mushroom and Yeast Beta-Glucan kit (K-YBGL) by excluding the alpha-glucans content from the total-glucans. As results, most of the extracts showed cytotoxicity on the lung cancer cells, A549 cells, rather than on the other cells, such as HepG2 (N=9 for each treatment). The extracts of reishi mushroom in 60% of methanol (dose range, 0.21-2.3 mg/ml) showed the strongest cytotoxicity on the lung cells, IC50=0.90 +/- 0.01 mg/ml, among the extracts of different methanol contents (10-60%) or different temperatures (50-90 °C) of hot water. However, the 60 % of methanol reishi extracts included relatively low levels of beta-glucans, compared to other extracts (18.57 +/- 0.31 vs. 22.42 +/- 5.56 % w/w). In addition, the levels of beta-glucans in all of reishi mushroom extracts were lower than those in all of the shiitake extracts (17.70 +/- 1.89 % vs. 27.05 +/- 4.21 % w/w: p<0.01). Moreover, there was no association between the levels of beta-glucans and IC50 in A549 cells. Interestingly, the combination (1:1) of the above 60% of methanol extracts of reishi mushroom and the 30% methanol extracts of shiitake (dose range, 0.30-3.26 mg/ml) showed synergic effects, i.e., 2-7 fold stronger effects (average of IC50=0.40 mg/ml) than every single treatment (average of IC50=0.90-2.63 mg/ml). Taken together, the reishi mushroom may have other strong chemopreventive materials rather than beta-glucans, while the shiitake may have a potentiation effect, when combined with the reishi mushroom. In conclusion, our 3D cell culture system showed high throughput performance to find the right preparation and effective usage of the two mushrooms and provided useful information for the next clinical studies for lung cancer.
Citation Format: Mihi Yang, Hee-Kyung Jang, Huiwon Kang. How do shiitake and reishi mushrooms work on lung cancer?: A high throughput screening of 3D cell culture. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4553.
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