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首页> 外文期刊>Cancer biology & therapy >Identification of genes downregulated in tumor cells expressing antisense glutaminase mRNA by differential display.
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Identification of genes downregulated in tumor cells expressing antisense glutaminase mRNA by differential display.

机译：通过差异显示鉴定在表达反义谷氨酰胺酶mRNA的肿瘤细胞中下调的基因。

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摘要

Ehrlich ascites tumor cells (EATC) is a highly proliferative malignant cell line derived from mouse mammary epithelia, whereas their derivative, 0.28AS-2 cells, expressing antisense glutaminase mRNA, show a less transformed phenotype and loss of their tumorigenic capacity in vivo correlated with an inhibition of glutaminase expression. The mRNA differential display technique was applied to these two cell lines for the identification and isolation of genes whose transcription was altered. Side-by-side comparisons of cDNA patterns among relevant RNA samples revealed four genes significantly downregulated in 0.28AS-2 cells: high-mobility group Hmga2 protein, Fmnl3 or formin-like protein 3, Nedd-4 ubiquitin-protein ligase, and ubiquitin carboxyl-terminal hydrolase Usp-15. These positives were confirmed by Northern analysis. The four targeted genes have relevant functions in cell growth and proliferation. Our results show the validity of mRNA differential display technique to get insights into the molecularmechanisms underlying the acquisition of a more differentiated phenotype by tumor cells after inhibition of glutaminase expression.
机译：Ehrlich腹水肿瘤细胞（EATC）是源自小鼠乳腺上皮细胞的高度增殖性恶性细胞系，而其表达反义谷氨酰胺酶mRNA的衍生物0.28AS-2细胞表现出较少的转化表型，并且在体内的致瘤能力丧失与抑制谷氨酰胺酶的表达。将mRNA差异显示技术应用于这两个细胞系，以鉴定和分离转录被改变的基因。对相关RNA样品中cDNA模式进行的并排比较显示，在0.28AS-2细胞中四个基因被显着下调：高迁移率族Hmga2蛋白，Fmnl3或FORMIN样蛋白3，Nedd-4泛素蛋白连接酶和泛素羧基末端水解酶Usp-15。这些阳性通过Northern分析得到证实。这四个靶向基因在细胞生长和增殖中具有相关功能。我们的结果表明，mRNA差异显示技术的有效性可用于深入了解谷氨酰胺酶表达抑制后肿瘤细胞获得更高分化表型的分子机制。

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