摘要
目的研究高迁移率蛋白N2(HMGN2)对人膀胱癌细胞T24及其裸鼠移植瘤的抑制作用。方法在体外细胞水平上,用噻唑蓝(MTT)法检测加入0,3,5,7,9μg·m L^(-1)HMGN2的细胞活性,用流式细胞仪检测0,1,3,5μg·m L^(-1)HMGN2对T24细胞的抑制作用;用Western blot法检测细胞中凋亡蛋白Bcl-2、胱天蛋白酶(caspase)-3、p53及Bax的表达。体内动物水平上,建立裸鼠移植瘤模型,分为空白组、对照组和实验组,分别注射磷酸盐缓冲液(PBS)、顺铂(DDP)和HMGN2于瘤体周边。20 d后处死裸鼠,取出瘤块,用苏木精-伊红(HE)染色及原位末端标记法(TUNEL)检测各组瘤组织的坏死及调亡。结果MTT结果表明,3,5,7,9μg·m L^(-1)HMGN2抑制了T24细胞的生长,且随着HMGN2浓度的增加及孵育时间的延长抑制率明显增加。流式细胞仪检测结果表明,1,3,5μg·m L^(-1)HMGN2对T24细胞总凋亡率分别为(13. 18±0. 83)%,(25. 79±1. 22)%,(31. 48±1. 82)%,与0μg·m L^(-1)HMGN2相比,差异有统计学意义(P <0. 01)。裸鼠移植瘤实验中,瘤组织的HE染色及TUNEL检测结果表明,与空白组相比,实验组和对照组瘤组织出现明显的坏死及凋亡。结论HMGN2可能通过促进瘤组织的坏死及诱导凋亡的作用抑制了T24细胞的增殖和裸鼠移植瘤的生长。
Objective The inhibitory effect of high mosomal protein N2 （HMGN2） on growth of human cells and xenograft tumor in nude mice. Methods mobility group chro-bladder cancer I＂24 In vitro, using thiazole blue method （MTY） to detect the effect of 0, 3, 5, 7, 9 p,g · mL^-1 HMGN2 on the activity of T24 cells. Flow cytometry was used to detect the inhibitory effect of 0, 1, 3, 5 μg· mL^-1 HMGN2 on T24 cells. The expressions of apoptotic proteins Bcl - 2, caspase - 3, p53 and Bax in ceils were detected by Western blot. In vivo, xenograft tumor in nude mice was constructed and divided into blank group, control group and ex- perimental group. Phosphate buffer solution （PBS）, cis - dichlorodia- mineplatinum （DDP） and HMGN2 were respectively injected around the tumor tissue. All nude mice were sacrificed on post- transplantation 20 d, tumor tissue was removed. The effects of necrosis and apoptosis was analyzed by Hematoxylin and eosin （HE） staining and TUNEL. Restilts MTY results indicated that the growth of T24 cells was inhibited by 3, 5, 7, 9μg ·mL^-1 HMGN2, and the inhibition rate increased significantly with the increase of HMGN2 concentration and the incubation time. Flow cytometry results showed that the total apoptosis rate of 1, 3, 5 μg · mL^- 1HMGN2 in T24 cells were （ 13.18± 0. 83 ） %, （25.79 ± 1.22） %, （ 31.48±1.82 ） %, had significant difference with 0 μg · mL-^1 HMGN2 （P 〈0. 01 ）. HE staining and TUNEL detection results of tumor tissues in nude mouser showed that compared with blank group, tumor tissues in experimental group and control group showed obvious necrosis and apoptosis. Conclusion HMGN2 may inhibit the proliferation of T24 ceils and the growth of xenograft tumor in nude mice by promoting the necrosis of tumor tissue and inducing apoptosis.
作者
蒋萍
岳进巧
李静
董震宇
杨旭
雷冬玉
吴桂霞
JIANG Ping;YUE Jin-qiao;LI Jing;DONG Zhen-yu;YANG X;LEI Dong-yu;WU Gui-xia(Department of Physiology,School of Basic Medical Sciences;Clinical Medicine,Xirtjiang Medical University,Urumqi 830011,China;Department of Medical Examination,Changii Vocational and Technical College,Changji 831100,Xinjiang Province,China;Department of Gynaecology and Obstetrics,the First Affiliated Hospital,Xinfiang Medical University,Urumqi 830011,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2018年第20期2402-2405,共4页
The Chinese Journal of Clinical Pharmacology
基金
国家自然科学基金资助项目(81660480)
新疆维吾尔自治区高校科研计划基金资助项目(XJEDU2014I019)
新疆昌吉职业技术学院科研计划基金资助项目(CJZY2017027)
关键词
高迁移率蛋白N2
T24细胞
移植瘤
膀胱癌
high mobility group chromosomal protein N2
T24 cell
xenograft tumor
carcinoma of urinary bladder
