Hypoxia-induced TUFT1 promotes the growth and metastasis of hepatocellular carcinoma by activating the Ca2+/PI3K/AKT pathway
Hypoxia-induced TUFT1 promotes the growth and metastasis of hepatocellular carcinoma by activating the Ca2+/PI3K/AKT pathway
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缺氧诱导的TUFT1通过激活Ca(2)/PI3K/AKT通路促进肝细胞癌的生长和转移。
DOI：
10.1038/s41388-018-0505-8
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发表时间：
2019-02-21
期刊：
影响因子：
8
通讯作者：
Tu, Kangsheng
中科院分区：
文献类型：
--
作者：
Dou, Changwei;Zhou, Zhenyu;Tu, Kangsheng
Tuftelin1 （TUFT1）是牙齿组织发育和矿化的酸性蛋白成分，受缺氧和Hedgehog信号通路的调节。我们研究了TUFT1在肝细胞癌（HCC）中的作用。采用qRT-PCR、免疫组化和western blot检测肝癌组织中TUFT1水平。采用MTT、BrdU、3D培养和Transwell试验评估细胞活力、增殖、体外生长、迁移和侵袭。建立皮下和尾静脉注射模型，观察其体内生长和转移情况。染色质免疫沉淀法评估缺氧诱导因子1 α (HIF-1 α)与TUFT1启动子的结合。使用microRNA阵列来鉴定与缺氧相关的microRNA。TUFT1在HCC中升高，与不良的临床病理特征和较差的生存率相关。在体外和体内，TUFT1通过激活Ca2+/PI3K/AKT通路促进HCC细胞生长、转移和上皮-间质转化。缺氧诱导TUFT1以HIF-1 α依赖的方式表达，HCC组织中TUFT1的表达与HIF-1a水平呈正相关。低氧通过下调miR-671-5p而不是直接促进HIF-1 α与TUFT1启动子的结合来增强TUFT1的表达。MiR-671-5p与TUFT1 mRNA的3'-UTR相互作用，随后抑制TUFT1的表达。因此，敲低TUFT1阻断了缺氧促进HCC进展的作用。TUFT1通过激活Ca2+/PI3K/AKT通路促进HCC细胞的生长、转移和EMT。低氧微环境通过下调miR-671-5p增加TUFT1的表达。TUFT1可能作为干预和治疗HCC的潜在治疗靶点。
Tuftelin1 (TUFT1), an acidic protein constituent of developing and mineralizing tooth tissues, is regulated by hypoxia and the Hedgehog signaling pathway. We investigated the role of TUFT1 in hepatocellular carcinoma (HCC). qRT-PCR, immunohistochemistry and western blot were employed to evaluate TUFT1 level in HCC. MTT, BrdU, 3D culture and Transwell assays were used to assess cell viability, proliferation, in vitro growth, migration, and invasion. Subcutaneous and tail vein injection models were established to investigate in vivo growth and metastasis. Chromatin immunoprecipitation was performed to assess binding of hypoxia-inducible factor 1 alpha (HIF-1 alpha) to TUFT1 promoter. A microRNA array was used to identify hypoxia-related microRNAs. TUFT1 was elevated in HCC, and correlated with unfavorable clinicopathologic characteristics and poor survival. TUFT1 promoted HCC cell growth, metastasis and epithelial-mesenchymal transition in vitro and in vivo via activation of Ca2+/PI3K/AKT pathway. Hypoxia induced TUFT1 expression in an HIF-1 alpha dependent manner, and TUFT1 expression was positively correlated with HIF-1a level in HCC tissues. Hypoxiaenhanced TUFT1 expression by downregulating miR-671-5p rather than by directly promoting the binding of HIF-1 alpha to TUFT1 promoter. MiR-671-5p interacted with the 3'-UTR of TUFT1 mRNA and subsequently inhibited TUFT1 expression. Consequently, knockdown of TUFT1 blocked the effects of hypoxia in promoting HCC progression. TUFT1 promoted the growth, metastasis and EMT of HCC cells through activating Ca2+/PI3K/AKT pathway. The hypoxic microenvironment increased the expression of TUFT1 via downregulation of miR-671-5p. TUFT1 may function as a potential therapeutic target for the intervention and treatment of HCC.