Jia Zhi-Fan, Pu Pei-Yu, Kang Chun-Sheng, Wang Guang-Xiu, Zhang Zhi-Yong, Qiu Ming-Zhe, Huang Qiang
Department of Neurosurgery, Tianjin Medical University General Hospital, Tianjin Neurological Institute and Laboratory of Neuro-Oncology, Tianjin, China.
Zhonghua Wai Ke Za Zhi. 2007 Oct 15;45(20):1420-3.
To investigate the influence of SEPT7 on biological characters of gliomas cells TJ905.
Recombinant SEPT7 constructs was transfected to human glioblastoma cell line TJ905 in which SEPT7 expression is absent. The positive clones were identified by RT-PCR and Western blot analysis. The cell proliferation was determined by MTT assay and flow cytometry, cell apoptosis was detected with Annexin V staining and cell invasion was evaluated by motility in three-dimensional culture. Moreover, the molecules regulating the cell cycle progression were examined by immunofluorescence staining and Western blot analysis.
When SEPT7 was successfully transfected to TJ905 cells, the cell proliferation activity of TJ905 cell was inhibited, the cell cycle was arrested in G0/G1 phase and S phase fraction (SPF) was lowered, the positive regulatory molecules for cell cycle progression including cyclin D1, CDk4, cyclin E and CDk2 were downregulated while the negative modulators including p16 and p21 were upregulated, apoptotic cells were increased and cell invasive ability was attenuated.
Transfection of SEPT7 construct into the glioma cells TJ905 is able to inhibit the proliferation activity and invasive ability of TJ905 cell and to induce cell apoptosis. These results revealed that SEPT7 exerted the suppressive effect on the glioma cell growth and invasion, and induced apoptosis, and suggested that SEPT7 as a gene of glioma suppressor.
研究SEPT7对胶质瘤细胞TJ905生物学特性的影响。
将重组SEPT7构建体转染至SEPT7表达缺失的人胶质母细胞瘤细胞系TJ905。通过RT-PCR和蛋白质免疫印迹分析鉴定阳性克隆。采用MTT法和流式细胞术检测细胞增殖,用膜联蛋白V染色检测细胞凋亡,通过三维培养中的运动性评估细胞侵袭。此外,通过免疫荧光染色和蛋白质免疫印迹分析检测调节细胞周期进程的分子。
当SEPT7成功转染至TJ905细胞时,TJ905细胞的增殖活性受到抑制,细胞周期停滞在G0/G1期,S期细胞分数(SPF)降低,细胞周期进程的正调控分子包括细胞周期蛋白D1、细胞周期蛋白依赖性激酶4、细胞周期蛋白E和细胞周期蛋白依赖性激酶2下调,而负调控分子包括p16和p21上调,凋亡细胞增加,细胞侵袭能力减弱。
将SEPT7构建体转染至胶质瘤细胞TJ905能够抑制TJ905细胞的增殖活性和侵袭能力,并诱导细胞凋亡。这些结果表明SEPT7对胶质瘤细胞的生长和侵袭具有抑制作用,并诱导凋亡,提示SEPT7作为一种胶质瘤抑制基因。