REIC/Dkk-3 stable transfection reduces the malignant phenotype of mouse prostate cancer RM9 cells
REIC/Dkk-3 stable transfection reduces the malignant phenotype of mouse prostate cancer RM9 cells
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DOI：
10.3892/ijmm_00000293
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发表时间：
2009-12-01
影响因子：
5.4
通讯作者：
Kumon, Hiromi
中科院分区：
文献类型：
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作者：
Chen, Jie;Watanabe, Masami;Kumon, Hiromi
在永生化细胞（REIC）/Dickkopf（Dkk）-3中表达减少。Dkk基因家族的成员，是多种癌症中的肿瘤抑制因子。建立了REIC/Dkk-3转染的小鼠前列腺癌RM 9细胞的稳定克隆（RM 9-REIC）和空载体转染的对照克隆细胞（RM 9-EV）。克隆用于评估抗癌效果，并进行REIC/Dkk-3连续表达的蛋白质组学分析。RM 9-REIC细胞表现出虚弱的外观，并且细胞膜显示出称为泡的不规则芽。与对照相比，RM 9-REIC克隆的体外细胞增殖被显著抑制。在标准培养条件下进行细胞凋亡测定，RM 9-REIC显示出较高的细胞凋亡发生率。将RM 9-EV和RM 9-REIC细胞原位植入C57 BL/6小鼠前列腺中。2周后，与对照相比，RM 9-REIC细胞中的肿瘤生长被显著抑制。双向凝胶电泳检测基因转染对蛋白表达的影响。质谱分析表明，过氧化物酶-1，GST-P1。RM 9-REIC细胞中transgelin-2、MRP-L12、ARD、GRP 78和Sorcin蛋白表达增加，eEF 1A-1和亲环素-40蛋白表达减少。因此，REIC/Dkk-3稳定转染子在体外和体内显示出小鼠前列腺癌RM 9细胞中恶性程度的降低。蛋白质组学分析结果可能为阐明REIC/Dkk-3基因转移的抗癌分子机制提供重要线索。
The reduced expression in immortalized cells (REIC)/Dickkopf (Dkk)-3. a member of the Dkk gene family, is a tumor suppressor in a broad range of cancers. REIC/Dkk-3 transfected stable clones of mouse prostate cancer RM9 cells (RM9-REIC) and the empty vector-transfected control clone cells (RM9-EV) were established. Clones were used to evaluate the anti-cancer effects and a proteomics analysis of REIC/Dkk-3 continuous expression was performed. The RM9-REIC cells show a feeble appearance and the cell membrane shows irregular buds known as blebs. In vitro cell proliferation was significantly suppressed in RM9-REIC clones in comparison to the control. The apoptosis assay was done under standard culture conditions and RM9-REIC showed a higher incidence of apoptosis. The RM9-EV and RM9-REIC cells were orthotopically implanted into a C57BL/6 mouse prostate. After 2 weeks, the tumor growth was significantly inhibited in RM9-REIC cells in comparison to the control. Two-dimensional gel electrophoresis was used to examine the modification of protein expression by the gene transfection. The analysis with mass spectrometry disclosed that expression of peroxiredoxin-l, GST-P1. transgelin-2, MRP-L12, ARD, GRP78 and Sorcin were increased and eEF1A-1 and cyclophilin-40 protein were decreased in RM9-REIC cells. Therefore, REIC/Dkk-3 stable transfectants show a reduction of malignancy in mouse prostate cancer RM9 cells in vitro and in vivo. The result of the proteomics analysis might provide important clues to clarify the anti-cancer molecular mechanism of REIC/Dkk-3 gene transfer.