GLI2 promotes cell proliferation and migration through transcriptional activation of ARHGEF16 in human glioma cells.
GLI2 promotes cell proliferation and migration through transcriptional activation of ARHGEF16 in human glioma cells.
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GLI2 通过转录激活人神经胶质瘤细胞中的 ARHGEF16 促进细胞增殖和迁移
DOI：
10.1186/s13046-018-0917-x
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发表时间：
2018-10-11
期刊：
影响因子：
--
通讯作者：
Luo S
中科院分区：
文献类型：
--
作者：
Huang D;Wang Y;Xu L;Chen L;Cheng M;Shi W;Xiong H;Zalli D;Luo S
Hedgehog（Hh）信号通路在调节胚胎发生和维持组织稳态中起着关键作用，其中胶质瘤相关癌基因（GLI）转录因子是主要介导者。该通路的异常激活与包括胶质母细胞瘤在内的各种人类恶性肿瘤相关，尽管其机制细节尚不清楚。我们进行了基因微阵列分析，这些基因在过表达GLI 2A（GLI 2的活性形式）的胶质母细胞瘤U87细胞中相对于对照细胞差异表达。染色质免疫沉淀和双荧光素酶测定用于确定Rho鸟嘌呤核苷酸交换因子16（ARHGEF 16）是否是GLI 2的下游靶标。然后，采用transwell迁移、EdU和软琼脂集落形成试验来测试ARHGEF 16对胶质瘤癌细胞迁移和增殖的影响，并在体内检查GLI 2/ARHGEF 16信号传导对肿瘤生长的影响。最后，我们进行了酵母双杂交试验，Co-IP和GST-下拉，以确定介导ARHGEF 16效应的因素。我们发现，ARHGEF 16 mRNA水平上调U87细胞过度表达GLI 2A相对于对照细胞。GLI 2与ARHGEF 16启动子结合并激活基因转录。与对照细胞相比，过表达ARHGEF 16的胶质瘤细胞U87和U118显示出增强的迁移和增殖，而与对照H4细胞相比，H4细胞中ARHGEF 16的敲低导致细胞增殖降低。与GLI 2A过表达对胶质瘤异种移植物生长的促进作用相反，GLI 2抑制和ARHGEF 16敲低均延缓肿瘤生长。细胞凋亡相关蛋白5（CKAP 5）是ARHGEF 16的相互作用蛋白，在ARHGEF 16促进胶质瘤细胞迁移和增殖中起重要作用。这些结果表明，靶向GLI 2/ARHGEF 16/CKAP 5信号传导轴的治疗策略可以抑制胶质瘤的进展和复发。本文的在线版本（10.1186/s13046-018-0917-x）包含补充材料，可供授权用户使用。
The Hedgehog (Hh) signaling pathway plays critical roles in modulating embryogenesis and maintaining tissue homeostasis, with glioma-associated oncogene (GLI) transcription factors being the main mediators. Aberrant activation of this pathway is associated with various human malignancies including glioblastoma, although the mechanistic details are not well understood. We performed a microarray analysis of genes that are differentially expressed in glioblastoma U87 cells overexpressing GLI2A, the active form of GLI2, relative to the control cells. Chromatin immunoprecipitation and dual-luciferase assays were used to determine whether Rho guanine nucleotide exchange factor 16 (ARHGEF16) is a downstream target of GLI2. Then, transwell migration, EdU and soft-agar colony formation assays were employed to test effects of ARHGEF16 on glioma cancer cell migration and proliferation, and the effects of GLI2/ARHGEF16 signaling on tumor growth were examined in vivo. Finally, we performed yeast two-hybrid assay, Co-IP and GST-pull down to identify factors that mediate effects of ARHGEF16. We found that ARHGEF16 mRNA level was upregulated in U87 cells overexpressing GLI2A relative to control cells. GLI2 binds to the ARHGEF16 promoter and activates gene transcription. Glioma cells U87 and U118 overexpressing ARHGEF16 showed enhanced migration and proliferation relative to the control cells, while knockdown of ARHGEF16 in H4 cells led to decreased cell proliferation compared to the control H4 cells. In contrast to the promoting effect of GLI2A overexpression on glioma xenograft growth, both GLI2 inhibition and ARHGEF16 knockdown retarded tumor growth. Cytoskeleton-associated protein 5 (CKAP5) was identified as an interaction protein of ARHGEF16, which is important for the stimulatory effects of ARHGEF16 on glioma cell migration and proliferation. These results suggest that therapeutic strategies targeting the GLI2/ARHGEF16/CKAP5 signaling axis could inhibit glioma progression and recurrence. The online version of this article (10.1186/s13046-018-0917-x) contains supplementary material, which is available to authorized users.
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影响因子：
10.5
作者：
Gergely, F;Draviam, VM;Raff, JW
通讯作者：
Raff, JW
影响因子：
3.9
作者：
Johnston, Jennifer J.;Sapp, Julie C.;Turner, Joyce T.;Amor, David;Aftimos, Salim;Aleck, Kyrieckos A.;Bocian, Maureen;Bodurtha, Joann N.;Cox, Gerald F.;Curry, Cynthia J.;Day, Ruth;Donnai, Dian;Field, Michael;Fujiwara, Ikuma;Gabbett, Michael;Gal, Moran;Graham, John M., Jr.;Hedera, Peter;Hennekam, Raoul C. M.;Hersh, Joseph H.;Hopkin, Robert J.;Kayserili, Hulya;Kidd, Alexa M. J.;Kimonis, Virginia;Lin, Angela E.;Lynch, Sally Ann;Maisenbacher, Melissa;Mansour, Sahar;McGaughran, Julie;Mehta, Lakshmi;Murphy, Helen;Raygada, Margarita;Robin, Nathaniel H.;Rope, Alan F.;Rosenbaum, Kenneth N.;Schaefer, G. Bradley;Shealy, Amy;Smith, Wendy;Soller, Maria;Sommer, Annmarie;Stalker, Heather J.;Steiner, Bernhard;Stephan, Mark J.;Tilstra, David;Tomkins, Susan;Trapane, Pamela;Tsai, Anne Chun-Hui;Van Allen, Margot I.;Vasudevan, Pradeep C.;Zabel, Bernhard;Zunich, Janice;Black, Graeme C. M.;Biesecker, Leslie G.
通讯作者：
Biesecker, Leslie G.
影响因子：
15.9
作者：
Dolecek, Therese A.;Propp, Jennifer M.;Kruchko, Carol
通讯作者：
Kruchko, Carol
影响因子：
56.9
作者：
Berman, DM;Karhadkar, SS;Beachy, PA
通讯作者：
Beachy, PA
DOI：
10.1158/1078-0432.ccr-13-2195
发表时间：
2014-03-15
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Huang L;Walter V;Hayes DN;Onaitis M
通讯作者：
Onaitis M