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. 2017 Jun 20;37(6):827-832.
doi: 10.3969/j.issn.1673-4254.2017.06.20.

[Mechanism of Platycarya strobilacea Sieb. et Zucc extract-induced methuosis in human nasopharyngeal carcinoma CNE1 and CNE2 cells]

[Article in Chinese]
Jun-Yu Zhu  1 , Wei Tu, Chao Zeng, Heng-Xu Mao, Qing-Feng DU, Hong-Bing Cai
Affiliations

[Mechanism of Platycarya strobilacea Sieb. et Zucc extract-induced methuosis in human nasopharyngeal carcinoma CNE1 and CNE2 cells]

[Article in Chinese]
Jun-Yu Zhu et al. Nan Fang Yi Ke Da Xue Xue Bao. .

Abstract
in English, Chinese

Objective: To study the effect of Platycarya strobilacea Sieb. et Zucc (PSZ) extract on methuosis of human nasopharyngeal carcinoma CNE1 and CNE2 cells and explore the underlying mechanism.

Methods: CNE1 and CNE2 cells were treated with 1 mg/mL PSZ extract and the expressions of Rac1 mRNA and Rac1 protein were detected using RT-qPCR and Western blotting, respectively. Results CNE1 and CNE2 cells showed obvious morphological changes typical of methuosis following treatment with PSZ extract characterized by cell merging, accumulation of large cytoplasmic vacuoles, and membrane rupture without obvious changes in the nuclei. PSZ treatment resulted in up-regulated Rac1 mRNA and Rac1 protein expressions in the cells. Application of EHT 1864 obviously blocked the effect of PSZ extract in inducing methuosis in CNE1 and CNE2 cells.

Conclusion: PSZ extract can induce methuosis in CNE1 and CNE2 cells by inducing the overexpression of Rac1.

目的: 探讨化香树果序乙醇提取物（PSZ）诱导鼻咽癌细胞CNE1、CNE2发生methuosis死亡的机制。

方法: 用1.0 mg/mL PSZ处理CNE1、CNE2细胞，观察细胞形态学变化，RT-qPCR、Western blotting检测PSZ干预后Rac1基因和蛋白表达水平。EHT 1864抑制两组细胞Rac1表达，观察化香树果序醇提物对CNE1、CNE2细胞的作用。

结果: CNE1、CNE2细胞经PSZ处理后，细胞质出现大量空泡，空泡相互融合，不断增大，最后细胞膜破裂，细胞核没有发生明显的变化，这种形态学改变符合methuosis细胞死亡方式，Rac1在基因和蛋白水平表达均增加。EHT 1864抑制细胞Rac1表达后，PSZ引起的methuosis死亡现象消失。

结论: Rac1是PSZ诱导人鼻咽癌细胞CNE1、CNE2发生methuosis死亡的关键靶点。

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Figures

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PSZ的HPLC图 HPLC diagram of the ethanol extract of PSZ.
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CNE1、CNE2细胞形态学变化 Morphology of CNE1 and CNE2 cells under inverted microscope (Original magnification: × 400). A: CNE1 control group; B: CNE1+PSZ (1 mg/mL, 24 h); C: CNE2 control group; D: CNE2+ PSZ (1 mg/mL, 24 h).
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PSZ处理组电镜结果 Morphological change of CNE cells treated with 1.0 mg/mL PSZ for 24 h observed with electron microscopy (×8000).
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荧光定量PCR检测Rac1 mRNA表达情况 Rac1 mRNA expression detected by fluorescent real-time PCR in CNE1 cells. *P < 0.05 vs CNE1 control group, #P < 0.05 vs CNE2 control group.
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PSZ作用后CNE1、CNE2细胞Rac1蛋白表达水平变化 Expression Rac1 in CNE1 and CNE2 cells after PSZ treatment. A: The bands of Western blotting; B: Ratio of the expression of Rac1. *P < 0.05 vs CNE1 control group, #P < 0.05 vs CNE2 control group.
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EHT1864干预后CNE1、CNE2细胞形态学变化 Morphology of CNE1 and CNE2 cells under inverted microscope with PSZ (1 mg/mL) and/or EHT1864 (20 μm) treatment for 24 h (×400).
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广东省自然科学基金（2016A030313724）；广东省中医药局科研项目（20173028）

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