Effect of exogenous S100A6 on human osteosarcoma cell line U2OS proliferation,apoptosis and β-catenin expression

CHEN Ying-hua; et al

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Effect of exogenous S100A6 on human osteosarcoma cell line U2OS proliferation,apoptosis and β-catenin expression

更新时间：2024-01-17

- Effect of exogenous S100A6 on human osteosarcoma cell line U2OS proliferation,apoptosis and β-catenin expression
- Journal of Chongqing Medical University Vol. 34, Issue 7, Pages: 817-821(2009)
- 作者机构：
  
  1. 重庆医科大学生物医学工程系、重庆市生物医学工程学重点实验室、重庆市超声医学工程重点实验室—省部共建国家重点实验室培育基地
  2. 美国芝加哥大学医学中心分子肿瘤研究室
- 作者简介：
- 基金信息：
- DOI：
  CLC： R738.1
- Published：2009，
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[1]陈英华,卫佳,李星星,吴丽美,马闻,张彦,何通川,周兰.外源性S100A6对人骨肉瘤细胞株U2OS的增殖、凋亡及β-catenin表达的影响[J].重庆医科大学学报,2009,34(07):817-821.

CHEN Ying-hua, et al. Effect of exogenous S100A6 on human osteosarcoma cell line U2OS proliferation,apoptosis and β-catenin expression. [J]. Journal of Chongqing Medical University 34(7):817-821(2009)
[1]陈英华,卫佳,李星星,吴丽美,马闻,张彦,何通川,周兰.外源性S100A6对人骨肉瘤细胞株U2OS的增殖、凋亡及β-catenin表达的影响[J].重庆医科大学学报,2009,34(07):817-821. DOI：

CHEN Ying-hua, et al. Effect of exogenous S100A6 on human osteosarcoma cell line U2OS proliferation,apoptosis and β-catenin expression. [J]. Journal of Chongqing Medical University 34(7):817-821(2009) DOI：

摘要

目的:探讨外源性S100A6对骨肉瘤细胞株U2OS的生物学作用及其可能机制。方法:MTT法检测S100A6对U2OS细胞增殖的影响及浓度依赖性

台盼蓝拒染法计数活细胞检测S100A6对U2OS增殖作用的时间依赖性

Hoechst染色检测细胞凋亡

Westernblot法及免疫细胞化学法检测细胞内β-catenin的表达变化。结果:S100A6作用3d时

浓度为30、100、300和1000μg/ml的重组S100A6（GST-S100A6）组的OD值分别比相应浓度的GST对照组的OD值减少20.5%、23.7%、32.2%和36.8%

P<0.05;蛋白浓度为100μg/ml时

GST-S100A6组的细胞数在前2d与GST组相似

第3、4、5d的细胞数分别降低到GST组的59%、46%、48%

P<0.05;GST-S100A6干预3d后细胞凋亡率增高3.16倍

P<0.01;免疫细胞化学法测得经GST-S100A6处理3d后β-catenin的平均光密度值比GST组增加22.9%

P<0.01;经携带S100A6基因的重组腺病毒（Ad-S100A6）处理3d后

Westernblot法检测细胞中β-catenin的校正灰度值为1.4204±0.1999

比对照组（Ad-GFP组

0.9996±0.1000）显著增高

P<0.01。结论:外源性S100A6能够抑制U2OS细胞的增殖、促进其凋亡和增加U2OS细胞中β-catenin水平。

Abstract

Objective: we investigated the biological effects of exogenous S100A6 on osteosarcoma cell line U2OS and explored its possible mechanism. Methods:MTT assay was used to determine S100A6 influence on U2OS cells proliferation and their conce ntration-dependent relationship. The trypan blue exclusion assay was used to observe S100A6 inhibitory effect on U2OS cells proliferation in a time-dependent manner;Hoechst staining was used to determine cell apoptosis;Western blot and immunocytochemical methods to determine expressions of cyto-β-catenin. Results:We found that

on the first three days

OD values in cells subjected to the recombinant protein S100A6(GST-S100A6)with the concentrations of 30

100

300 and 1 000μg/ml respectively were obviously lower than those in the GST group

by 20.5%

23.7%

32.2% and 36.8% accordingly(P <0.05);when protein concentration was 100 μg/ml

cell counting in GST-S100A6 group was similar with GST group in the first two days

while decreased significantly to 59%

46%

48% on the third

fourth

fifth day compared with the GST group (P <0.05);cell apoptosis rates increased by 3.16 fold three days after subjected to GST-S100A6(P <0.01). Average optical density ofβ-catenin in GST-S100A6 group increased by 22.9% compared with the GST group (P <0.01)72h after exposure

using immunocytochemical assay;72 h after treatment with the recombinant adenovirus carrying S100A6 gene (Ad-S100A6)

corrected gray value of intracellular β-catenin was 1.420 4±0.199 9

obviously higher than those in the control group 0.999 6±0.100 0 (P<0.01).Conclusion:Exogenous protein S100A6 could inhibite U2OS cells proliferation

promote apoptosis and then increase intracellularβ-catenin concentration.

关键词

S100A6骨肉瘤β-catenin

Keywords

S100A6osteosarcomaβ-catenin

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