Department of Neurosurgery, The Affiliated Huai'an Hospital of Xuzhou Medical University, The Second People's Hospital of Huai'an, Huai'an 223002, China.
Department of Neurosurgery, The Second Affiliated Hospital of Xuzhou Medical University, Xuzhou 221006, China.
Int J Biol Macromol. 2021 Apr 15;176:145-156. doi: 10.1016/j.ijbiomac.2021.02.043. Epub 2021 Feb 8.
Karyopherins mediate the macromolecular transport between the cytoplasm and the nucleus and participate in cancer progression. However, the role and mechanism of importin-11 (IPO11), a member of the karyopherin family, in glioma progression remain undefined. Effects of IPO11 on glioma progression were detected using CCK-8, colony formation assay, flow cytometry analysis, caspase-3 activity assay, and Transwell invasion assay. Western blot analysis was used to detect the expression of active caspase-3, active caspase-7, active caspase-9, N-cadherin, Vimentin, E-cadherin, β-catenin, and c-Myc. The activity of Wnt/β-catenin pathway was evaluated by the T-cell factor/lymphoid enhancer factor (TCF/LEF) transcription factor reporter assay. Results showed that IPO11 knockdown inhibited proliferation and reduced colony number in glioma cells. IPO11 silencing promoted the apoptotic rate, increased expression levels of active caspase-3, caspase-7, and caspase-9, and enhanced caspase-3 activity. Moreover, IPO11 silencing inhibited glioma cell invasion by suppressing epithelial-to-mesenchymal transition (EMT). Mechanistically, IPO11 knockdown inactivated the Wnt/β-catenin pathway. β-Catenin overexpression abolished the effects of IPO11 silencing on the proliferation, apoptosis, and invasion in glioma cells. Furthermore, IPO11 silencing blocked the malignant phenotypes and repressed the Wnt/β-catenin pathway in vivo. In conclusion, IPO11 knockdown suppressed the malignant phenotypes of glioma cells by inactivating the Wnt/β-catenin pathway.
载体蛋白将细胞质和细胞核之间的大分子运输，并参与癌症的进展。然而，载体蛋白家族的一员 Importin-11（IPO11）在神经胶质瘤进展中的作用和机制仍未确定。使用 CCK-8、集落形成实验、流式细胞术分析、半胱天冬酶-3 活性测定和 Transwell 侵袭实验检测 IPO11 对神经胶质瘤进展的影响。Western blot 分析用于检测活性半胱天冬酶-3、活性半胱天冬酶-7、活性半胱天冬酶-9、N-钙黏蛋白、波形蛋白、E-钙黏蛋白、β-连环蛋白和 c-Myc 的表达。通过 T 细胞因子/淋巴增强因子（TCF/LEF）转录因子报告实验评估 Wnt/β-连环蛋白通路的活性。结果表明，IPO11 敲低抑制神经胶质瘤细胞的增殖并减少集落数量。IPO11 沉默促进了细胞凋亡率，增加了活性半胱天冬酶-3、半胱天冬酶-7 和半胱天冬酶-9 的表达水平，并增强了半胱天冬酶-3 的活性。此外，IPO11 沉默通过抑制上皮间质转化（EMT）抑制神经胶质瘤细胞的侵袭。机制上，IPO11 敲低使 Wnt/β-连环蛋白通路失活。β-连环蛋白过表达消除了 IPO11 沉默对神经胶质瘤细胞增殖、凋亡和侵袭的影响。此外，IPO11 沉默在体内阻断了恶性表型并抑制了 Wnt/β-连环蛋白通路。总之，IPO11 敲低通过使 Wnt/β-连环蛋白通路失活抑制了神经胶质瘤细胞的恶性表型。
