外源性S100A8在体外对Hela细胞具有抑制性作用

基础医学与临床 ›› 2011, Vol. 31 ›› Issue (10): 1082-1087.

外源性S100A8在体外对Hela细胞具有抑制性作用

郭元元¹,游莉²,²,徐兰兰²,²,邹正渝²,²,黎玉叶²,²,孙双双²,²,罗进勇²,²,周兰³

1. 重庆医科大学临床检验诊断学教育部重点实验室
2.
3. 重庆医科大学

收稿日期:2010-11-11 修回日期:2011-03-04 出版日期:2011-10-05 发布日期:2011-10-08
通讯作者: 周兰 E-mail:zhoulan0111@gmail.com
基金资助:
国家重点基础研究发展计划

The inhibitive effects of exogenous S100A8 on Hela cells in vitro

Received:2010-11-11 Revised:2011-03-04 Online:2011-10-05 Published:2011-10-08

摘要/Abstract

摘要： 目的：探讨外源性S100A8对宫颈癌细胞系Hela的增殖、凋亡、克隆形成及迁移和侵袭的影响。方法：MTT法检测细胞增殖活性；Hoechst染色检测细胞凋亡；流式细胞技术检测细胞周期变化；平板克隆形成实验检测细胞克隆形成能力；划痕(Wound healing)实验检测细胞的迁移能力；Transwell侵袭实验检测细胞的侵袭能力。结果:1）抑制Hela细胞增殖：MTT显示细胞培养3d时, 浓度为100 mg/L、300 mg/L和1000 mg/L的GST-hS100A8组的OD值较GST组减少13.64％、19.29％和25.06％(P<0.05)；在浓度为100 mg/L时，GST-hS100A8组OD值的减少也呈一定时间依赖性，从第3d起差异具有统计学意义（P<0.05）；2）促进Hela细胞凋亡：Hoechst染色显示GST-hS100A8组在第3d时细胞凋亡率较GST组增加5.18倍（P<0.05），同时，流式细胞仪分析也发现其在第3d出现峰值为19.9%的凋亡峰，而对照组没有凋亡峰；3）对Hela细胞的周期没有明显影响；4）抑制Hela细胞的克隆形成：GST-hS100A8组的克隆形成率较GST组减少30.2%（P＜0.05）；5）抑制Hela细胞迁移：划痕实验显示在24h时GST-hS100A8组的划痕愈合率较GST组降低30.1%（P＜0.05）；6）抑制Hela细胞侵袭能力：Transwell显示GST-hS100A8组在24h时穿膜细胞数较GST组减少48.9%（P＜0.05）。结论: 外源性S100A8能够抑制Hela细胞的增殖、克隆形成、迁移和侵袭能力并诱导凋亡，提示S100A8可能具有抑制宫颈癌的作用。

关键词: S100A8, Hela, 抑制

Abstract: Objective To study the effect of exogenous S100A8 on cell proliferation，apoptosis, clone formation, migration and invasion of human cervical cancer cell line Hela. Methods MTT was used to detect the cell proliferation; Hoechst staining was used to detect the cell apoptosis; Flow Cytometry was used to detect the variation of cell cycle; colony-forming assay was used to detect the ability of cell cloning formation; Wound healing assay and Transwell chamber experiment were used to detect the cell migration and invasion ability. Results 1)S100A8 inhibited cell proliferation: MTT indicated that after treatment of GST-hS100A8 for 3d, the OD value of 100 mg/L, 300 mg/L and 1000 mg/L group of GST-hS100A8 decreased by 13.64%, 19.29% and 25.06% compared with GST group, respectively (P＜0.05) .In the group of 100 mg/L, the OD value of GST-hS100A8 decreased in a time-dependent manner, especially from 72h(P＜0.05). 2)S100A8 promoted cell apoptosis: The result of Hoechst staining showed that cell apoptosis rate in GST-hS100A8 group increased by 5.18 times compared with GST group at 72h(P＜0.05). Meanwhile, the result of Flow Cytometry displayed that the peak of apoptosis of GST-hS100A8 was 19.9% at 72h, while the groups of GST and blank were no apoptotic peak. 3) We also found that the effect of GST-hS100A8 on cell cycle was not significant. 4) Colony-forming assay showed that the rate of cloning formation of GST-hS100A8 decreased by 30.2% (P＜0.05). 5) The result of wound healing assay indicated that the healing rate of GST-hS100A8 group reduced by 30.1% compared with GST group (P＜0.05) at 24h. 6) The result of Transwell indicated that the trans-membrane cell number of GST-hS100A8 group reduced by 48.9%(P＜0.05) compared with GST group at 24h. Conclusion Exogenous S100A8 could inhibit cell proliferation, cloning formation, migration, and invasion and induce cell apoptosis on human cervical cancer cell line Hela, which indicated that S100A8 could have the inhibitive effects on cervical cancer.

Key words: S100A8, Hela, Inhibitive effect

中图分类号:

R73-3

郭元元游莉徐兰兰邹正渝黎玉叶孙双双罗进勇周兰. 外源性S100A8在体外对Hela细胞具有抑制性作用[J]. 基础医学与临床, 2011, 31(10): 1082-1087.

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