Expression and transcriptional profiling of the LKB1 tumor suppressor in cervical cancer cells.
Expression and transcriptional profiling of the LKB1 tumor suppressor in cervical cancer cells.
复制标题
LKB1 肿瘤抑制因子在宫颈癌细胞中的表达和转录谱。
DOI：
10.1016/j.ygyno.2014.04.050
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发表时间：
2014-08
影响因子：
4.7
通讯作者：
Chen JJ
中科院分区：
文献类型：
--
作者：
Zhang X;Chen H;Wang X;Zhao W;Chen JJ
为了研究LKB1的生物学活性，检测LKB1蛋白的表达，寻找可能作为宫颈癌治疗靶点的LKB1调控基因。检测表达LKB1的宫颈癌HeLa细胞的增殖情况。采用免疫组织化学方法检测LKB1在正常宫颈组织和宫颈癌组织中的表达。用基因芯片技术分析稳定表达LKB1的宫颈癌HeLa细胞的基因表达谱。使用基因本体论(GO)术语和京都基因和基因组百科全书(KEGG)途径数据库分析差异表达的基因。采用定量RT-PCR方法对芯片数据进行验证。Western blotting检测到脂质磷酸酶肌醇多磷酸4-磷酸酶(INPP4B)的表达。LKB1的表达抑制了HeLa细胞的增殖，激活了AMPK，在超过50%的宫颈癌中缺失。在有无LKB1的HeLa细胞中，有200多个基因有差异表达。GO注释的生物信息学分析表明，LKB1在接受各种刺激并将其转化为分子信号方面发挥了作用。KEGG通路分析表明，有8条通路受到显著调控。这些代谢包括精氨酸和脯氨酸代谢以及肌醇磷酸代谢。随机选择的7个基因进行定量RT-PCR，证实了差异表达。此外，在LKB1过表达的细胞中，INPP4B蛋白的稳态水平上调。这项研究确立了LKB1是宫颈癌中一个重要的肿瘤抑制因子，并揭示了LKB1调控的一条新的信号通路。
To characterize the biological activities of LKB1, examine LKB1 protein expression and identify LKB1-regulated genes that may serve as therapeutic targets in cervical cancer. Proliferation of cervical cancer HeLa cells expressing LKB1 was examined. LKB1 expression in normal cervical tissues and cervical cancers was assessed by immunohistochemistry. Gene expression profiles of cervical cancer HeLa cells stably expressing LKB1 were analyzed by microarray. Differentially expressed genes were analyzed using Gene Ontology (GO) terms and the Kyoto Encyclopedia of Genes and Genomes (KEGG) PATHWAY database. Quantitative RT-PCR was used to validate the microarray data. The expression of lipid phosphatase inositol polyphosphate 4-phosphatase type II (INPP4B) was confirmed by western blotting. Expression of LKB1 inhibited HeLa cell proliferation and activated AMPK and was lost in more than 50% of cervical carcinomas. More than 200 genes were differentially expressed between HeLa cells with and without LKB1. Bioinformatics analysis with GO annotation indicated that LKB1 plays a role in receiving diverse stimuli and converting them into molecular signals. KEGG PATHWAY analysis showed that 8 pathways were significantly regulated. These include arginine and proline metabolism and inositol phosphate metabolism. The differential expression of 7 randomly selected genes was confirmed by quantitative RT-PCR. Furthermore, the steady-state level of INPP4B protein was up-regulated in LKB1-overexpressing cells. This study establishes LKB1 as an important tumor suppressor in cervical cancer and sheds light on a novel signaling pathway regulated by LKB1.