Malignant phenotype of renal cell carcinoma cells is switched by Ukrain administration in vitro
Malignant phenotype of renal cell carcinoma cells is switched by Ukrain administration in vitro
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DOI：
10.1097/cad.0b013e328346c7f7
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发表时间：
2011-09
影响因子：
2.3
通讯作者：
N. Gagliano;L. Pettinari;M. Aureli;C. Martinelli;E. Colombo;F. Costa;Roberta Carminati;Tatiana Volpari;G. Colombo;A. Milzani;I. Dalle-Donne;M. Gioia
中科院分区：
文献类型：
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作者：
N. Gagliano;L. Pettinari;M. Aureli;C. Martinelli;E. Colombo;F. Costa;Roberta Carminati;Tatiana Volpari;G. Colombo;A. Milzani;I. Dalle-Donne;M. Gioia
我们研究了Ukrain是否调节透明细胞肾细胞癌（ccRCC）细胞Caki-1、Caki-2和ACHN在4种剂量（5、10、20和40 mol/l）作用24和48小时后的恶性表型。通过免疫荧光以及肌动蛋白和微管蛋白分析了上皮到间质转化标记E-cadherin、-catenin和vimentin；采用sds酶谱分析基质金属蛋白酶-2和基质金属蛋白酶-9活性，western blot分析细胞内和分泌的SPARC水平，流式细胞术分析细胞周期。Ukrain虽然能影响Caki-2和ACHN中肌动蛋白皮层的表达，但不能在细胞-细胞边界诱导E-cadherin/-catenin的免疫反应性，也不影响vimentin的组织；然而，在一些Caki-1和ACHN细胞中，vimentin的核周浓度与其下调一致。给药20 mol/l后48 h，基质金属蛋白酶-2和基质金属蛋白酶-9活性显著下调。在这个时间点上，在Caki-2和Caki-1和ACHN细胞中，在20 mol/l剂量下，乌克兰显著减少了细胞迁移和侵袭，并下调了细胞上清中SPARC的水平。同时，SPARC在所有ccRCC细胞中都上调，这表明乌克兰也可能通过细胞周期抑制来影响细胞增殖，正如细胞周期分析所支持的那样，因为SPARC也可以作为细胞周期抑制剂。我们的研究结果表明，Ukrain可能会改变ccRCC细胞上皮向间质转化相关的表型，并针对与RCC进展有关的两个主要方面，如肿瘤侵袭/微环境重塑和细胞增殖。
We investigated whether Ukrain modulates the malignant phenotype of clear cell renal cell carcinoma (ccRCC) cells Caki-1, Caki-2, and ACHN treated with four doses (5, 10, 20, and 40 mol/l) for 24 and 48 h. The epithelial-to-mesenchymal transition markers E-cadherin, -catenin, and vimentin were analyzed by immunofluorescence as well as actin and tubulin; matrix metalloproteinase-2 and matrix metalloproteinase-9 activity was analyzed by SDS-zymography, intracellular and secreted SPARC levels by western blot, and cell cycle by flow cytometry. Ukrain did not induce E-cadherin/-catenin immunoreactivity at the cell–cell boundary, although it determined the actin cortical expression in Caki-2 and ACHN, and did not affect vimentin organization; however, in some Caki-1 and ACHN cells the perinuclear concentration of vimentin was consistent with its downregulation. Matrix metalloproteinase-2 and matrix metalloproteinase-9 activity was significantly downregulated 48 h after 20 mol/l Ukrain administration. At this time point, Ukrain significantly decreased migration and invasion, and downregulated SPARC levels in cell supernatants at all doses in Caki-2, and at 20 mol/l in Caki-1 and ACHN cells. Concomitantly, SPARC was upregulated in all ccRCC cells, suggesting that Ukrain could also affect cell proliferation by cell cycle inhibition, as supported by the cell cycle analysis, as SPARC also acts as a cell cycle inhibitor. Our results suggest that Ukrain may switch the epithelial-to-mesenchymal transition-related phenotype of ccRCC cells, and targets the two major aspects involved in RCC progression, such as tumor invasion/microenvironment remodeling and cell proliferation.