摘要
目的以人口腔鳞状细胞癌细胞株Tca8113为实验模型,初步探讨高迁移率族蛋白N2(HMGN2)抗口腔鳞状细胞癌的作用。方法大量培养重组人HMGN2表达载体大肠杆菌BL21,用异丙基-1-硫代-β-呋喃半乳糖苷(IPTG)诱导HMGN2的表达,产物用B-PER GST Fusion Protein Purification Kit进行纯化。在细胞培养基中加入不同质量浓度的HMGN2,通过MTT、Hoechst 33342荧光染色、流式细胞术和Western-blot法检测其凋亡效果及抗凋亡的分子机制。结果经MTT检测证明HMGN2能显著抑制Tca8113细胞的生长,通过Hoechst 33342荧光染色、流式细胞术和Western-blot检测证明HMGN2能使Tca8113的细胞形态发生改变,并且能使Tca8113细胞阻滞于细胞周期的S期,促进Tca8113细胞凋亡。结论 HMGN2可以促进人口腔鳞状细胞癌细胞的凋亡。
Objective Take human oral squamous cell carcinoma Tca8113 as experimental model, and study the anti oral squamous cell carcinoma activity of high mobility group chromosomal protein N2 （HMGN2） molecule. Methods Train a large number of recombinant human HMGN2 expression vector Escherichia coli BL21. HMGN2 was expressed under isopropyl-l-thio-β-galactopyranoside（IPTG） induction and purified by B-PER GST Fusion Protein Purification Kit. A variety of concentrations HMGN2 were added to cell culture medium, cells were tested by MrlT, Hoeehst 33342 fluorescence staining, flow cytometry assay and Western-blot. Results MTT results proved that HMGN2 could signifi- cantly inhibit human oral squamous cell carcinoma Tca8113 growth. Hoechst 33342 fluorescence staining, flow cytome- try assay test and Western-blot proved HMGN2 could make Tca8113 cells morphological change, make Tea8113 ceils block in S period of cell cycle and strongly promote Tca8113 cells to apoptosis. Conclusion HMGN2 can promote apoptosis of oral squamous cell carcinoma cells.
出处
《华西口腔医学杂志》
CAS
CSCD
北大核心
2013年第1期91-95,共5页
West China Journal of Stomatology
基金
国家自然科学基金资助项目(30972764)
关键词
高迁移率族蛋白N2
人口腔鳞状细胞癌细胞株
凋亡
high mobility group chromosomal protein N2~ human oral squamous cell carcinoma cell line
apop- tosis
