Search for the genes regulated by Histone deaetylase inhibitor and the Rb gene

HDAC抑制剂，如FK 228和阿司他丁A（TSA），诱导多种人类癌细胞的生长停滞和凋亡。我们在20种人肺癌细胞系（包括15种非小细胞肺癌（NSCLC）和5种小细胞肺癌（SCLC）细胞系）中暴露于FK 228或TSA 72 h后，通过MTS测定和FACS分析评价细胞毒性。为了研究肺癌细胞对FK 228、TSA和9种化疗药物（顺铂、CBDCA、VP-16、SN-38、紫杉醇、多西他赛、长春瑞滨、吉西他滨、SM 5887）敏感性的基因表达谱，我们应用基因芯片技术分析了20个肺癌细胞对FK 228、TSA和9种化疗药物敏感性的基因。应用于表达数据的聚类算法显示HDAC抑制剂和9种化疗剂之间的区分亚组。此外，我们将表达数据与细胞系的IC 50数据进行了比较。我们发现感觉和情绪之间 ...更多信息 HDAC抑制剂的活性和包括CST 3在内的几个基因的表达水平，但未能通过实时PCR方法证实它们的显著关联。视网膜母细胞瘤肿瘤抑制蛋白（RB）在大多数人类肿瘤（包括NSCLC和SCLC）中被靶向灭活。RB通过抑制细胞周期进程所必需的基因的转录来抑制增殖，并且已经显示Rb基因恢复到SCLC细胞中可以抑制体外致瘤性。为了抑制转录，RB通过直接结合E2 F并组装含有染色质修饰酶（包括组蛋白脱乙酰酶（HDAC））的多蛋白复合物来功能性拮抗E2 F活性。然而，HDAC参与转录抑制的程度以及RB介导的抑制所需的程度尚不清楚。由于HDAC抑制剂可诱导非小细胞肺癌和小细胞肺癌细胞凋亡。我们研究了Rb基因重建肺癌细胞是否阻断HDAC诱导的凋亡途径。HDAC抑制剂与作为对照的腺病毒-Rb或腺病毒-LacZ的组合处理通过组合指数法类似地显示出加和至轻度协同效应，表明Rb的重建没有阻断HDAC抑制剂诱导的细胞凋亡。少

HDAC inhibitors, such as FK228 and trichostatin A(TSA), induce growth arrest and apoptosis in a variety of human cancer cells. We evaluated cytotoxicity by MTS assay and FACS analysis after 72 h of exposure to FK228 or TSA in 20 human lung cancer cell lines including 15 non-small cell lung cancer(NSCLC) and 5 small cell lung cancr(SCLC) cell lines. We found both FK228 and TSA steeply inhibited cell growth and induced apoptosis in 20 cell lines with nanomolar concentrations.To investigate genes to sensitivity of lung cancers cells to FK228, TSA and 9 chemotherapeutic agents including CDDP, CBDCA, VP-16, SN-38 Paclitaxel, Docetaxel, Vinorelbine, Gemcitabine, SM5887, we performed cDNA microarray analysis of 20 lung cancer cells. A clustering algorithm applied to the expression data showed distinguished subgroups between HDAC inhibitors and 9 chemotherapeutic agents. In addition, we compared the expression data with IC50 data of cell lines. We found significant associations between sensiti … More vity to HDAC inhibitors and expression levels of several genes including CST3, but failed to confirm their significant association by real-time PCR method.The retinoblastoma tumor suppressor protein(RB) is targeted for inactivation in the majority of human tumors, including NSCLC and SCLC. RB inhibits proliferation by repressing the transcription of genes that are essential for cell cycle progression and restoration of the Rb gene to SCLC cells has been shown to suppress the tumorigenecity in vitro. To repress transcription, RB functionally antagonize E2F acitivity through binding E2Fs directly and assembling multiprotein complexes containing chromatin-modifying enzymes, including histone deacetylases(HDACs). However, the extent to which HDACs participate in transcriptional repression and are required for RB-mediated repression is unclear. Since the HDAC inhibitors induced apoptosis in NSCLC and SCLC cells. We examined whether reconstitution of the Rb gene to lung cancer cells blocks the HDAC-induced apoptotic pathway. Combined treatment of HDAC inhibitors with Adenovirus-Rb or Adenovirus-LacZ as a control similarly showed additive to mild synergistic effects by combination index method, indicating reconstitution of Rb did not block the HDAC inhibitor-induced apoptosis. Less