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14 June 2014 : Original article

TESTIN suppresses tumor growth and invasion via manipulating cell cycle progression in endometrial carcinoma

Zhenpeng GuC, Guofeng DingCD, Kuixiang LiangAEF, Hongtao ZhangBD, Guanghong GuoDE, Lili ZhangBC, Jinxiu CuiCDE

DOI: 10.12659/MSM.890544

Med Sci Monit 2014; 20:980-987

Abstract

BACKGROUND: 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.

MATERIAL AND METHODS: 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.

RESULTS: 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.

CONCLUSIONS: 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.

Keywords: Animals, Cytoskeletal Proteins - metabolism, Endometrial Neoplasms - pathology, Endometrium - pathology, Gene Knockdown Techniques, LIM Domain Proteins - metabolism, Mice, Inbred BALB C, Mice, Nude, Xenograft Model Antitumor Assays

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750