ARHGEF10L contributes to liver tumorigenesis through RhoA-ROCK1 signaling and the epithelial-mesenchymal transition
ARHGEF10L contributes to liver tumorigenesis through RhoA-ROCK1 signaling and the epithelial-mesenchymal transition
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ARHGEF10L 通过 RhoA-ROCK1 信号传导和上皮间质转化促进肝脏肿瘤发生
DOI：
10.1016/j.yexcr.2018.11.007
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发表时间：
2019-01-01
影响因子：
3.7
通讯作者：
Chang, Xiaotian
中科院分区：
文献类型：
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作者：
Tang, Junyi;Liu, Chunyan;Chang, Xiaotian
Rho小G蛋白及其调节因子的异常活性在肿瘤的发生发展中起着重要作用。Rho鸟嘌呤核苷酸交换因子10L(ARHGEF10L)是促进Rho GTP酶活性结合状态的Rhogef家族成员之一。本研究使用Illumina GoldenGate微量分析、Sequenom Massarray和TaqMan分析ARHGEF10L基因座的Tag单核苷酸多态(Tag SNPs)与各种肿瘤风险的可能相关性。基因分型分析显示rs2244444和rs12732894与肝癌有很强的相关性。免疫组织化学和Western blotting也显示ARHGEF10L在肝细胞癌组织中的表达增加。此外，转染ARHGEF10L表达载体后，肝癌细胞株Bel-7402和HepG2细胞的增殖、迁移和RhoA活性增加，Rho相关螺旋卷曲蛋白-1(ROCK1)、磷酸化Ezrin/Radiin/Moesin(ERM)、波形蛋白、N-钙粘蛋白和slug的表达增加，E-钙粘蛋白的表达降低。反义ArHGEF10L siRNA在两种细胞系中的表达结果相反。用包装短发夹状ARHGEF10L RNA的慢病毒载体转染Bel-7402细胞，建立荷瘤小鼠。ARHGEF10L表达受抑的裸鼠移植瘤生长减少，Vimentin、N-钙粘附素和Slug的表达减少。此外，在Bel-7402和HepG2细胞中，反义ROCK1 siRNA表达下调，而在肝细胞癌组织中ROCK1表达增强。E-钙粘蛋白、波形蛋白、N-钙粘蛋白和Slug是上皮向间充质转化(EMT)的标志。ROCK1、磷酸化ERM和EMT已被报道促进肿瘤细胞的增殖、转移和血管生成。我们的研究表明，ARHGEF10L的高表达通过激活RhoA-ROCK1-磷酸化ERM途径和EMT来刺激肝细胞肿瘤的发生。
Aberrant activity of Rho small G-proteins and their regulators plays an important role in tumorigenesis. Rho guanine nucleotide exchange factor 10-Like (ARHGEF10L) is a member of the RhoGEF family that promotes the active GTP-bound state of Rho GTPases. This study used the Illumina GoldenGate microassay, Sequenom MassARRAY and TaqMan to analyze possible correlations between tag single nucleotide polymorphisms (tag SNPs) in the ARHGEF10L locus and various tumor risks. The genotyping analyses demonstrated a strong association of rs2244444 and rs12732894 with liver cancer. Western blotting and immunohistochemistry also revealed increased expression of ARHGEF10L in hepatocellular carcinoma tissues. Furthermore, increased cell proliferation, cell migration and RhoA activity; increased expression of Rho-associated coiled-coil kinase-1 (ROCK1), phospho- Ezrin/Radixin/Moesin (ERM), vimentin, N-cadherin and Slug, and decreased E-cadherin expression were detected in hepatocellular carcinoma cell Bel-7402 and HepG2 cells with transfection of ARHGEF10L-expressing plasmids. Opposite results were obtained in the two cell lines with transfection of anti-ARHGEF10L siRNA. Tumor-bearing mice were generated with Bel-7402 cells transfected with lentivirus vectors packaging short hairpin ARHGEF10L RNA. The xenograft tumors with the inhibited ARHGEF10L expression showed decreased tumor growth and expression of vimentin, N-cadherin and Slug. Additionally, decreased phospho-ERM expression was detected in Bel-7402 and HepG2 cells with transfection of anti-ROCK1 siRNA and increased expression of ROCK1 was detected in hepatocellular carcinoma tissues. E-cadherin, vimentin, N-cadherin and Slug are markers of the epithelial-to-mesenchymal transition (EMT). ROCK1, phospho-ERM and EMT have been reported to promote tumor cell proliferation, metastasis and angiogenesis. Our study suggests that increased expression of ARHGEF10L stimulates hepatocellular tumorigenesis by activating the RhoA-ROCK1- phospho ERM pathway and EMT.