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Phyllanthus emblica Linn. fruit extract potentiates the anticancer efficacy of mitomycin C and cisplatin and reduces their genotoxicity to normal cells in vitro

余甘子提取物增强丝裂霉素C和顺铂的抗癌作用并降低其对正常细胞的遗传毒性

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Abstract

Objective

Fruit of Phyllanthus emblica Linn. (PE) is widely consumed as a functional food and used as a folk medicine due to its remarkable nutritional and pharmacological effects. Mitomycin C (MMC) and cisplatin (cDDP) are the most widely used forms of chemotherapeutic drug, but their clinical use is limited by their genotoxicity to normal cells. We aimed to determine whether PE has potential to reduce the genotoxicity, while improving the anticancer effect, of MMC and cDDP.

Methods

Cell proliferation was evaluated using the trypan blue exclusion assay and colony-forming assay. Genomic instability (GIN) was measured using the cytokinesis-block micronucleus assay.

Results

Co-treatment (72 h) with PE at 20–320 μg/ml significantly enhanced the efficacy of MMC (0.05 μg/ml) and cDDP (1 μg/ml) against Colo205 colorectal cancer cells (P<0.05), and at 80–320 μg/ml significantly decreased MMCand cDDP-induced GIN and multinucleation in normal colonic NCM460 cells (P<0.05). PE significantly decreased the mitotic index (P<0.01), blocked mitotic progression (P<0.05), and promoted apoptosis (P<0.01) in MMC- and cDDP-treated NCM460 cells, suggesting that PE-mediated inhibition of mitosis and induction of apoptosis may limit the division and survival of highly damaged cells. Also, PE was found to inhibit the clonal expansion of MMC- and cDDP-treated NCM460 cells (P<0.05) and decrease the heterogeneity of the surviving clones.

Conclusions

PE potentiates the anticancer efficacy of MMC and cDDP, while preventing their genotoxicity and inhibiting clonal expansions of unstable genomes in normal cells. These data suggest that PE has the potential to reduce the risk of secondary cancers induced by chemotherapeutics.

中文概要

目的

评估余甘子提取物(PE)对丝裂霉素C(MMC)和顺铂(cDDP)抗癌活性及其遗传毒性副作用的影响。

创新点

首次发现PE能减弱MMC和cDDP对人正常结肠上皮细胞基因组的损伤以及降低基因组受损细胞的克隆形成能力和克隆异质性。

方法

人结肠癌Colo205细胞和人正常结肠上皮NCM460细胞分别经PE、PE+MMC组合或PE+cDDP组合处理72 h。细胞增殖用台盼蓝拒染法和克隆形成法测定,遗传毒性用胞质分裂阻断微核分析法(CBMN)测定。

结论

结果显示,PE可以显著增强MMC和cDDP的抗Colo205细胞增殖能力(图1)。同时,PE显著降低MMC和cDDP诱导的NCM460细胞基因组不稳定现象,包括降低微核、核质桥和核芽(表1和图2)以及多核化细胞(图3)。此外,PE显著降低经MMC和cDDP处理的NCM460细胞克隆性扩增能力,并降低克隆的异质性(图4)。综上所述,PE不仅能增强MMC和cDDP的抗癌能力,还可能具有减弱它们诱发正常细胞恶性转变的潜力。

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Acknowledgements

This work was also supported by Yunnan Province New Academic Talent Award for Ph.D. 2016 (to Xi-han GUO).

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  1. The two authors contributed equally to this work

Authors and Affiliations

  1. School of Life Sciences, the Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Yunnan Normal University, Kunming, 650500, China

    Xi-han Guo, Juan Ni & Xu Wang

  2. School of Life Sciences, Yunnan University, Kunming, 650091, China

    Xi-han Guo & Xu Wang

  3. Institute of Genetics, Fudan University, Shanghai, 200433, China

    Jing-lun Xue

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  1. Xi-han Guo
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  2. Juan Ni
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Correspondence to Xu Wang.

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Project supported by the National Natural Science Foundation of China (Nos. 31260268 and 31560307)

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Guo, Xh., Ni, J., Xue, Jl. et al. Phyllanthus emblica Linn. fruit extract potentiates the anticancer efficacy of mitomycin C and cisplatin and reduces their genotoxicity to normal cells in vitro. J. Zhejiang Univ. Sci. B 18, 1031–1045 (2017). https://doi.org/10.1631/jzus.B1600542

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