Gu Zhenpeng, Ding Guofeng, Liang Kuixiang, Zhang Hongtao, Guo Guanghong, Zhang Lili, Cui Jinxiu
Department of Obstetrics and Gynecology, Affiliated Hospital of Binzhou Medical College, Binzhou, China (mainland).
Department of Infectious Diseases, Affiliated Hospital of Binzhou Medical College, Binzhou, China (mainland).
Med Sci Monit. 2014 Jun 14;20:980-7. doi: 10.12659/MSM.890544.
The TESTIN gene was demonstrated to be a tumor suppressor in prostate and breast cancer through inhibiting tumor growth and invasion. Herein, we aimed to investigate the detailed functions of TESTIN in the highly sexual hormone (estrogen)-dependent malignancy, endometrial carcinoma.
TESTIN mRNA and protein expression were measured by qRT-PCR, Western blot and immunohistochemistry. Upregulation of TESTIN was achieved by transfecting the pcDNA3.1-TESTIN plasmids into AN3CA cells. Knockdown of TESTIN was achieved by transfecting the shRNA-TESTIN into Ishikawa cells. MTT assay, colony formation assay, and Transwell assay were used to investigate the effects of TESTIN on cellular proliferation and invasion. The apoptotic status and cell cycle were analyzed using flow cytometry. MMP2 secretion was determined by ELISA assay. The xenograft assay was used to investigate the functions of TESTIN in nude mice.
Compared to the non-malignant adjacent endometrium, 54% of tumor samples presented downregulation of TESTIN (P<0.001). Loss of TESTIN protein was correlated with advanced tumor stage (P=0.047), high grade (P=0.034), and lymphatic vascular space invasion (P=0.036). In vitro, overexpression of TESTIN suppressed cell proliferation, induced dramatic G1 arrest, and inhibited tumor invasion through blocking the secretion of MMP2. Loss of TESTIN accelerated cellular proliferation, promoted cell cycle progression, and enhanced tumor invasion by increasing the secretion of MMP2. Consistently, TESTIN could significantly delay the growth of xenografts in nude mice.
TESTIN was commonly downregulated in human endometrial carcinoma and was associated with poor prognostic markers. Moreover, TESTIN significantly inhibited tumor growth and invasion via arresting cell cycle in in vitro and in vivo experiments. Therefore, we propose that TESTIN might be a prognostic marker and therapeutic target for endometrial carcinoma.
TESTIN基因已被证明通过抑制肿瘤生长和侵袭，在前列腺癌和乳腺癌中发挥肿瘤抑制作用。在此，我们旨在研究TESTIN在高度依赖性激素（雌激素）的恶性肿瘤——子宫内膜癌中的详细功能。
采用qRT-PCR、蛋白质免疫印迹法和免疫组织化学法检测TESTIN mRNA和蛋白表达。通过将pcDNA3.1-TESTIN质粒转染至AN3CA细胞来上调TESTIN表达。通过将shRNA-TESTIN转染至Ishikawa细胞来敲低TESTIN表达。采用MTT法、集落形成试验和Transwell试验研究TESTIN对细胞增殖和侵袭的影响。使用流式细胞术分析细胞凋亡状态和细胞周期。通过ELISA法测定MMP2分泌。采用异种移植试验研究TESTIN在裸鼠中的功能。
与非恶性的相邻子宫内膜相比，54%的肿瘤样本中TESTIN表达下调（P<0.001）。TESTIN蛋白缺失与肿瘤晚期（P=0.047）、高级别（P=0.034）以及淋巴管间隙浸润（P=0.036）相关。在体外，TESTIN过表达抑制细胞增殖，诱导显著的G1期阻滞，并通过阻断MMP2分泌抑制肿瘤侵袭。TESTIN缺失加速细胞增殖，促进细胞周期进程，并通过增加MMP2分泌增强肿瘤侵袭。同样，TESTIN可显著延缓裸鼠体内异种移植瘤的生长。
TESTIN在人类子宫内膜癌中普遍下调，并与不良预后标志物相关。此外，在体外和体内实验中，TESTIN通过阻滞细胞周期显著抑制肿瘤生长和侵袭。因此，我们认为TESTIN可能是子宫内膜癌的预后标志物和治疗靶点。
