Jin Hua, Zhang Xuexin, Su Jun, Teng Yueqiu, Ren Huan, Yang Lizhuang
Department of Immunology, Harbin Medical University, Harbin, Heilongjiang 150081, P.R. China.
Department of Neurosurgery, The Third Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150040, P.R. China.
Mol Med Rep. 2015 Nov;12(5):6617-25. doi: 10.3892/mmr.2015.4280. Epub 2015 Sep 2.
Translationally controlled tumor protein (TCTP) is a highly conserved, growth‑associated and small molecule protein, which is highly expressed in various types of tumor cell. TCTP can promote the growth and suppress apoptosis of tumor cels. However, few studies have reported the effects of TCTP in gliomas. In the present study, a glioma cell line was established, which was stably transfected with TCTP short hairpin ribonucleic acid (shRNA), to investigate the impact of downregulated expression of TCTP on the proliferation, apoptosis and invasion of glioma cells. Western blot and reverse transcription-quantitative polymerase chain reaction analyses demonstrated that TCTP shRNA effectively reduced the expression of TCTP in the U251 glioma cell line. MTT and colony formation assays revealed that downregulated expression of TCTP significantly inhibited glioma cell proliferation. Cell cycle analysis using flow cytometry revealed that the cells in the pRNA‑H1.1‑TCTP group were arrested in the G0/G1 phase of the cell cycle. Western blot analysis detected downregulated expression levels of cyclins, including Cyclin D1, Cyclin E and Cyclin B. Annexin V‑fluorescein isothiocyanate/propidium iodide and Hoechst staining demonstrated that the apoptotic rate of the cells in the pRNA‑H1.1‑TCTP group was significantly higher than that of the cells in the pRNA‑H1.1‑control group, with upregulated expression levels of B-cell-associated X protein and cleaved‑caspase‑3 and downregulated expression of B-cell lmyphoma-2 in the apoptotic process. Wound healing and Transwell assays revealed that downregulated expression of TCTP significantly inhibited the migration and invasiveness of the glioma cells; and the expression levels and activities of matrix metalloproteinase (MMP)‑2 and MMP‑9 were also significantly affected. In conclusion, the present study demonstrated that downregulated expression of TCTP significantly inhibited proliferation and invasion, and induced apoptosis in the glioma cells. These results suggested that TCTP may be important in glioma development and metastasis. Therefore, TCTP is expected to become an effective target for glioma gene therapy.
翻译调控肿瘤蛋白（TCTP）是一种高度保守、与生长相关的小分子蛋白，在各种类型的肿瘤细胞中高表达。TCTP可促进肿瘤细胞生长并抑制其凋亡。然而，关于TCTP在胶质瘤中的作用鲜有研究报道。在本研究中，建立了稳定转染TCTP短发夹核糖核酸（shRNA）的胶质瘤细胞系，以研究TCTP表达下调对胶质瘤细胞增殖、凋亡和侵袭的影响。蛋白质免疫印迹法（Western blot）和逆转录-定量聚合酶链反应分析表明，TCTP shRNA有效降低了U251胶质瘤细胞系中TCTP的表达。MTT法和集落形成试验显示，TCTP表达下调显著抑制了胶质瘤细胞的增殖。流式细胞术细胞周期分析显示，pRNA-H1.1-TCTP组细胞停滞于细胞周期的G0/G1期。蛋白质免疫印迹分析检测到细胞周期蛋白（包括细胞周期蛋白D1、细胞周期蛋白E和细胞周期蛋白B）的表达水平下调。膜联蛋白V-异硫氰酸荧光素/碘化丙啶及Hoechst染色表明，pRNA-H1.1-TCTP组细胞的凋亡率显著高于pRNA-H1.1-对照组，在凋亡过程中，B细胞相关X蛋白和裂解的半胱天冬酶-3表达上调，而B细胞淋巴瘤-2表达下调。划痕试验和Transwell试验显示，TCTP表达下调显著抑制了胶质瘤细胞的迁移和侵袭能力；基质金属蛋白酶（MMP）-2和MMP-9的表达水平及活性也受到显著影响。总之，本研究表明，TCTP表达下调显著抑制了胶质瘤细胞的增殖和侵袭，并诱导其凋亡。这些结果提示，TCTP可能在胶质瘤的发生发展和转移中起重要作用。因此，TCTP有望成为胶质瘤基因治疗的有效靶点。
