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Year : 2014  |  Volume : 10  |  Issue : 2  |  Page : 299-304

HMGN2 protein inhibits the growth of infected T24 cells in vitro

State Key laboratory of Oral Diseases, Sichuan University, Chengdu, China

Correspondence Address:
Yun Feng
State Key laboratory of Oral Diseases, Sichuan University, No 14, Section 3 South Renming Road, Chengdu, 610041
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0973-1482.136577

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Aims of Study: Natural killer (NK) cells and cytolytic T lymphocytes (CTL) have been implicated as important effectors of antitumor defense. High mobility group nucleosomal-binding domain 2 (HMGN2) may be one of the effector molecules of CTL and NK cells. The antitumor effect and mechanism of HMGN2 was investigated in this study. Materials and Methods: HMGN2 was isolated and purified from the human monocyte cell line THP-1 and then characterized by Tricine-SDS-PAGE, western blot, and mass spectrum determination. Confluent T24 cells were incubated with Klebsiella pneumoniae for 2 h, after which the extracellular bacteria were killed by the addition of gentamicin. The cells then were treated with a variety of concentrations of HMGN2. The effect of HMGN2 on the proliferation of T24 cells was analyzed with MTT, Hoechst and flow cytometry assays. Results: Cell growth assay results demonstrated that HMGN2 significantly inhibited the growth of T24 bladder cancer cell lines infected by K. pneumoniae. Furthermore, results of the Hoechst and flow cytometry assays indicated that HMGN2 may promote apoptosis in this experimental model. These results suggest HMGN2 could inhibit the growth of the infected human bladder cancer cells in vitro. Conclusion: HMGN2 protein could inhibit the growth of infected T24 cells in vitro, and the anti-tumor action of HMGN2 was due to induce apoptosis.

Abstract in Chinese

HMGN2蛋白抑制体外培养的T24细胞增殖 摘要 目的:自然杀伤(NK)细胞和细胞毒性T淋巴细胞(CTL)在肿瘤免疫方面有起重要作用。高迁移率族核小体结合结构域2(HMGN2)可能是CTL和NK细胞的效应分子,本实验主要研究HMGN2的抗肿瘤作用及机制。 材料与方法:从人单核细胞株THP 1分离和纯化HMGN2,并通过聚丙烯酰胺凝胶电泳(Tricine-SDS-PAGE),免疫印迹法,和质谱进行鉴定。融合T24细胞在肺炎克雷伯菌孵育2h后,其胞外菌再用庆大霉素杀死。不同浓度的HMGN2处理细胞。MTT,Hoechst染色和流式细胞仪检测HMGN2对T24cells增殖的影响。 结果:细胞生长测定结果表明,HMGN2有明显抑制受肺炎克雷伯菌感染膀胱癌T24细胞株的作用。此外, Hoechst染色和流式细胞仪检测结果表明,在本实验模型中HMGN2可以促进细胞凋亡。这些结果提示HMGN2可抑制体外感染人膀胱癌细胞的生长。 结论:HMGN2蛋白能够抑制感染T24细胞体外生长,HMGN2的抗肿瘤作用是由于诱导细胞凋亡。 关键词:抗肿瘤活性,细胞凋亡,高迁移率族核小体结合的结构域2,感染t24细胞

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