三氧化二砷对高迁移率族蛋白诱导的大鼠滑膜细胞增殖的抑制作用

doi:河北省科技厅自然科学基金资助课题（C 2007

三氧化二砷对高迁移率族蛋白诱导的大鼠滑膜细胞增殖的抑制作用

郭惠芳^1,2 ，刘淑霞¹，张玉军¹，左连富³，郭建文³，张欣³，张景坤¹，张悦¹

1.河北医科大学基础医学院病理学教研室，河北石家庄 050017；2.河北医科大学第二医院免疫风湿科，河北石家庄 050017；3.河北省肿瘤研究所流式分析室，河北石家庄 050011

收稿日期:2007-10-17 修回日期:1900-01-01 出版日期:2008-01-28 发布日期:2008-01-28
通讯作者: 左连富

Inhibitory effect of arsenic trioxide on proliferation of rat synoviocyte stimulated by HMGB1

GUO Hui-fang^1,2, LIU Shu-xia¹, ZHANG Yu-jun¹, ZUO Lian-fu², GUO Jian-wen³, ZHANG Xin³, ZHANG Jing-kun¹, ZHANG Yue¹

1.Department of Pathology, School of Basic Medical Sciences,Hebei Medical University, Shijiazhuang 050017，China；2.Department of Immunology and Rheumatology, Second Hospital, Hebei Medical University, Shijiazhuang 050017，China；3.Department of FCM Analysis，Hebei Provincial Tumor Institute，Shijiazhuang 050011，China

Received:2007-10-17 Revised:1900-01-01 Online:2008-01-28 Published:2008-01-28
Contact: ZUO Lian-fu

摘要/Abstract

摘要： 目的：探讨三氧化二砷（AS₂O₃）对高迁移率族蛋白1（HMGB1）诱导的大鼠滑膜细胞(RSC-364细胞)增殖的抑制作用及其机制。方法：将常规培养的RSC-364细胞分为正常对照组、HMGB1组及HMGB1加0、5、10、20、40和80 μmol•L^-1 AS₂O₃实验组； MTT法检测不同浓度AS₂O₃对RSC-364细胞生长的影响，RT-PCR检测STAT1 mRNA的表达变化，免疫细胞化学和流式细胞术检测STAT1、cyclin D1和增殖细胞核抗原（PCNA）蛋白表达的变化。结果：① MTT结果显示，AS₂O₃可抑制HMGB1诱导的RSC-364细胞的增殖，80　μmol•L^-1 AS₂O₃作用24　h抑制率可达88.18%，其抑制作用呈时间和浓度依赖性（P<0.05或P<0.01）；② 与对照组比较，HMGB1组STAT1 mRNA及STAT1、PCNA 、cyclin D1蛋白表达量增强，AS₂O₃组STAT1 mRNA及STAT1、PCNA、cyclin D1蛋白降低，并呈剂量依赖性(P<0.01）；③ STAT1与cyclin D1蛋白表达呈正相关关系（r=0.842，P=0.001）。结论：AS₂O₃能够抑制HMGB1诱导的滑膜细胞的增殖，其机制可能与抑制STAT1的活性从而下调细胞周期调节蛋白cyclin D1的表达有关。

关键词: 砷剂, 高迁移率族蛋白类, 信号转导和转录激活子1, 细胞周期蛋白D1

Abstract: Abstract:Objective To investigate the inhibitory effect of arsenic trioxid（AS₂O₃）on the proliferation of RSC-364 synoviocyte lines stimulated by HMGB1 and its possible mechanism. Methods RSC-364 synoviocytes were cultivated with standard medium as control group or with medium supplemented with HMGB1(HMGB1 group) or HMGB1 and 0, 5,10, 20, 40，80 μmol•L^-1 AS₂O₃(experimental groups),respectively. MTT assay was carried out to study the inhibitory effects of AS₂O₃ with different concentrations on the growth of RSC-364. RT-PCR was used to detect the mRNA expression of STAT1. Proteins of phospho-STAT1, cyclin D1 and proliferation cell nuclear antigen (PCNA) were detected by immunocytochemistry and FCM.Results ① MTT data indicated AS₂O₃ could inhibit the growth of RSC-364 stimulated by HMGB1 in a time-dependent and dose-dependent manner. ②Compared with control group, HMGB1 up-regulated the expression of phospho-STAT1 protein and mRNA as well as PCNA protein. Both mRNA and protein levels of phospho-STAT1 were decreased in the RSC-364 cells exposed to AS₂O₃ in a dose-dependent manner; AS₂O₃ also down-regulated the expression of PCNA protein. ③ HMGB1 up-regulated the expression of cyclin D1, while AS₂O₃ decreased the expression of cyclin D1 .④ There was positive correlation between phospho-STAT1 and cyclin D1 (r=0.842,P=0.001); Conclusion Arsenic trioxide could inhibit the proliferation of RSC-364 synoviocytes, partly by down-regulating the expression of phospho-STAT1/cyclin D1 .

Key words: arsenicals, high mobility group box chromosomal proteins, signal transducers and activators of transcription 1, cyclin D1

中图分类号:

R361.3

郭惠芳，刘淑霞，张玉军，左连富，郭建文，张欣，张景坤，张悦. 三氧化二砷对高迁移率族蛋白诱导的大鼠滑膜细胞增殖的抑制作用[J]. J4, 2008, 34(1): 108-111.

GUO Hui-fang, LIU Shu-xia, ZHANG Yu-jun, ZUO Lian-fu, GUO Jian-wen, ZHANG Xin, ZHANG Jing-kun, ZHANG Yue. Inhibitory effect of arsenic trioxide on proliferation of rat synoviocyte stimulated by HMGB1[J]. J4, 2008, 34(1): 108-111.

[1]	杨倩倩, 崔久嵬, 李薇, 陈耐飞, 王冠军. 三氧化二砷联合全反式维甲酸诱导治疗初治急性早幼粒细胞白血病患者疗效的影响因素分析[J]. J4, 2010, 36(3): 558-562.
[2]	郭亚雄, 刘艳波, 李德龙, 韩向北, 许多, 程宏, 赵丽晶, 赵丽娟, 董妍. 20(s)-原人参二醇对前列腺癌RM-1细胞的生长抑制作用及其机制[J]. J4, 2010, 36(2): 349-353.
[3]	于春艳, 康劲松, 李洪岩, 钟加滕, 王伟伟, 孙连坤. HIP1在维生素K3诱导Hela细胞损伤中的表达及意义[J]. J4, 2009, 35(5): 790-793.
[4]	刘玉和，于春艳，王皓，孙连坤. Vk3对宫颈癌Hela细胞mTOR信号转导蛋白基因表达的影响[J]. J4, 2009, 35(2): 267-270.
[5]	徐丹，杨幼林，王彦君. 三氧化二砷对人大肠癌细胞端粒酶活性及细胞凋亡的影响[J]. J4, 2008, 34(2): 195-198.
[6]	贾国瑜，王冠军，李薇. 三氧化二砷治疗急性早幼粒细胞白血病患者脑脊液砷浓度的测定及临床意义[J]. J4, 2007, 33(6): 1086-1087.