中华皮肤科杂志 ›› 2018, Vol. 51 ›› Issue (12): 892-896.doi: 10.3760/cma.j.issn.0412-4030.2018.12.009

• 研究报道 • 上一篇    下一篇

茶多酚对人乳头瘤病毒16亚基因永生化人宫颈上皮细胞生长的影响

莫子茵 陈荃 李华平 代歆悦 彭丽倩 尹忠浩 黄久遂 梁碧华 李润祥 李振洁 杨日东 朱慧兰   

  1. 510095 广州医科大学皮肤病研究所（莫子茵、尹忠浩）；广州市皮肤病防治所（陈荃、李华平、代歆悦、彭丽倩、黄久遂、梁碧华、李润祥、李振洁、杨日东、朱慧兰）
  • 收稿日期:2018-01-31 修回日期:2018-05-12 出版日期:2018-12-15 发布日期:2018-11-30
  • 通讯作者: 朱慧兰 E-mail:zhlhuilan@126.com
  • 基金资助:
    广东省自筹经费类科技计划项目（2017ZC0416）；广州市科技计划项目（201604020093、201804010058、201804010010）；广州市医药卫生科技项目（20171A011287）

Effect of tea polyphenols on the growth of human papillomavirus 16 subgenes-immortalized human cervical epithelial cells

Mo Ziyin, Chen Quan, Li Huaping, Dai Xinyue, Peng Liqian, Yin Zhonghao, Huang Jiusui, Liang Bihua, Li Runxiang, Li Zhenjie, Yang Ridong, Zhu Huilan   

  1. Institute of Dermatology, Guangzhou Medical University, Guangzhou 510095, China （Mo ZY, Yin ZH）; Guangzhou Institute of Dermatology, Guangzhou 510095, China （Chen Q, Li HP, Dai XY, Peng LQ, Huang JS, Liang BH, Li RX, Li ZJ, Yang RD, Zhu HL）
  • Received:2018-01-31 Revised:2018-05-12 Online:2018-12-15 Published:2018-11-30
  • Contact: Zhu Huilan E-mail:zhlhuilan@126.com
  • Supported by:
    Guangdong Self?financing Science and Technology Project （2017ZC0416）; Guangzhou Science and Technology Planning Project （201604020093, 201804010058, 201804010010）; Guangzhou Medical and Health Technology Project （20171A011287）

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摘要： 目的 探讨茶多酚对人乳头瘤病毒16（HPV16）亚基因永生化宫颈上皮细胞（H8细胞）生长的影响。方法 采用0（对照组）、6.25、12.5、25、50 mg/L茶多酚与H8细胞共孵育24、36、48 h后，CCK8法检测细胞增殖，孵育24 h后流式细胞仪检测细胞凋亡与细胞周期，荧光显微镜观察细胞凋亡形态。结果 不同浓度茶多酚孵育H8细胞24、36、48 h可抑制H8细胞的增殖，12.5 mg/L茶多酚可时间依赖性降低H8细胞的相对生长率。流式细胞仪检测结果示，0、6.25、12.5、25、50 mg/L茶多酚组细胞凋亡率分别为29.96% ± 0.70%、52.62% ± 0.62%、52.22% ± 0.72%、42.52% ± 0.90%、45.96% ± 2.11%，组间差异有统计学意义（F = 272.00，P < 0.05），各浓度茶多酚组细胞凋亡率均显著高于对照组（均P < 0.05）。荧光显微镜下可见茶多酚处理组H8细胞出现核固缩、核碎裂等典型凋亡形态学改变。各浓度组细胞G1、G2期细胞比例和细胞增殖指数差异均有统计学意义（P < 0.05）。与对照组相比，6.25、12.5、25 mg/L组G1期细胞比例增加（55.96% ± 0.72%、54.12% ± 3.20%、65.30% ± 1.51%，均P < 0.05），G2期细胞比例减少（3.17 ± 1.82%、4.94 ± 1.46%、4.65 ± 4.26%，均P < 0.05），增殖指数降低（0.44 ± 0.01、0.46 ± 0.02、0.36 ± 0.01，均P < 0.05）。结论 茶多酚可抑制H8细胞增殖，诱导细胞凋亡，阻滞细胞周期。

关键词: 儿茶素； 细胞系, 转化； 细胞增殖； 细胞凋亡； 细胞周期； 癌前状态； 茶多酚； 永生化人宫颈上皮细胞

Abstract: Mo Ziyin, Chen Quan, Li Huaping, Dai Xinyue, Peng Liqian, Yin Zhonghao, Huang Jiusui, Liang Bihua, Li Runxiang, Li Zhenjie, Yang Ridong, Zhu Huilan Institute of Dermatology, Guangzhou Medical University, Guangzhou 510095, China （Mo ZY, Yin ZH）; Guangzhou Institute of Dermatology, Guangzhou 510095, China （Chen Q, Li HP, Dai XY, Peng LQ, Huang JS, Liang BH, Li RX, Li ZJ, Yang RD, Zhu HL） Corresponding author: Zhu Huilan, Email: zhlhuilan@126.com 【Abstract】 Objective To explore the effect of tea polyphenols on the growth of human papillomavirus 16 （HPV16） subgenes-immortalized human cervical epithelial cells （H8 cells）. Methods Cultured H8 cells were divided into 5 groups to be treated with 0 （control group）, 6.25, 12.5, 25 and 50 mg/L tea polyphenols respectively for 24, 36, and 48 hours, and then cell counting kit-8（CCK8）assay was performed to detect cell proliferation. After 24 hours of incubation, flow cytometry was conducted to detect cell apoptosis and cell cycle, and fluorescence microscopy to observe the morphology of apoptotic cells. Results After incubation with tea polyphenols at different concentrations for 24, 36 and 48 hours, the proliferation of H8 cells was inhibited, and 12.5 mg/L tea polyphenols could inhibit the relative growth rate of H8 cells in a time-dependent manner. Flow cytometry showed that there was a significant difference in cell apoptosis rate among the 6.25-, 12.5-, 25-, 50-mg/L tea polyphenols groups and the control group（52.62% ± 0.62%, 52.22% ± 0.72%, 42.52% ± 0.90%, 45.96% ± 2.11%, 29.96% ± 0.70% respectively, F = 272.0, P < 0.05）. Moreover, all the tea polyphenol groups showed significantly increased cell apoptosis rate compared with the control group（all P < 0.05）. Fluorescence microscopy showed karyopyknosis, nuclear fragmentation and other typical apoptotic morphological changes in H8 cells in tea polyphenols groups. There were significant differences in the percentage of cells in G1, G2 phase and cell proliferation index among the 5 groups （all P < 0.05）. Compared with the control group, the 6.25-, 12.5-, 25-mg/L tea polyphenols groups showed significantly increased percentage of cells in G1 phase （55.96% ± 0.72%, 54.12% ± 3.20%, 65.30% ± 1.51% respectively, all P < 0.05）, but significantly decreased percentage of cells in G2 phase （3.17 ± 1.82%, 4.94 ± 1.46%, 4.65 ± 4.26% respectively, all P < 0.05） and lower cell proliferation index（0.44 ± 0.01, 0.46 ± 0.02, 0.36 ± 0.01 respectively, all P < 0.05）. Conclusion Tea polyphenols can inhibit the proliferation of H8 cells, induce cell apoptosis, and block cell cycle progression.

Key words: Catechin, Cell line, transformed, Cell proliferation, Apoptosis, Cell cycle, Precancerous conditions, Tea polyphenols, Immortalized human cervical epithelial cells