Department of Neurosurgery, Huazhong University of Science and Technology Union Shenzhen Hospital, The 6th Affiliated Hospital of Shenzhen University Health Science Center (Shenzhen Nanshan People's Hospital), Shenzhen 518056, Guangdong, China.
Department of Neurosurgery, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen, Guangdong, China.
J Mol Neurosci. 2021 Aug;71(8):1636-1647. doi: 10.1007/s12031-021-01832-8. Epub 2021 Apr 19.
The present study aimed to detect the effect of tenascin C (TNC) on cell function and chemosensitivity to paclitaxel and phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) signaling in glioma cells.Human glioma cells U87, LN-229, T98G and U251 and normal human astrocytes were obtained, in which TNC expression was detected. The U87 cells and U251 cells were chosen and infected with lentivirus of control overexpression, TNC overexpression, control knockdown, and TNC knockdown for functional experiments. Rescue experiments were then performed to evaluate the effect of PI3K/AKT activator 740 Y-P on cell function and chemosensitivity to paclitaxel in TNC knockdown U251 cells. TNC mRNA and protein expression was elevated in glioma cells, including U87, LN-229, U251 and T98G cells, compared to normal human astrocytes. In U87 and U251 cells, TNC promoted proliferation while inhibiting apoptosis. In addition, TNC upregulated PI3K and p-AKT protein expression in U87 and U251 cells. As for chemosensitivity, TNC increased relative viability in U251 cells treated with 400 ng/mL and 800 ng/mL paclitaxel. In terms of stemness, TNC increased the sphere number per 1000 cells, CD44CD133 cell percentage and 1/stem cell frequency (assessed by extreme limiting dilution analysis) in U251 cells. In rescue experiments, 740 Y-P reduced the effect of TNC on proliferation, apoptosis, chemosensitivity to paclitaxel, and stemness in U251 cells. TNC acts as an oncogenic factor by promoting cancer cell proliferation and stemness while inhibiting apoptosis and chemosensitivity to paclitaxel in glioma via modulation of PI3K/AKT signaling.
本研究旨在检测 tenascin C（TNC）对胶质瘤细胞的细胞功能和紫杉醇及磷脂酰肌醇 3-激酶/蛋白激酶 B（PI3K/AKT）信号通路的化学敏感性的影响。获得了人胶质瘤细胞 U87、LN-229、T98G 和 U251 以及正常人星形胶质细胞，并检测了 TNC 的表达。选择 U87 细胞和 U251 细胞，用对照过表达、TNC 过表达、对照敲低和 TNC 敲低的慢病毒进行功能实验。然后进行挽救实验，以评估 PI3K/AKT 激活剂 740 Y-P 对 TNC 敲低 U251 细胞的细胞功能和紫杉醇化学敏感性的影响。与正常人星形胶质细胞相比，TNC 在胶质瘤细胞（包括 U87、LN-229、U251 和 T98G 细胞）中的 mRNA 和蛋白表达水平升高。在 U87 和 U251 细胞中，TNC 促进增殖，同时抑制凋亡。此外，TNC 上调了 U87 和 U251 细胞中 PI3K 和 p-AKT 蛋白的表达。至于化学敏感性，TNC 增加了 U251 细胞中用 400ng/mL 和 800ng/mL 紫杉醇处理后的相对存活率。在干性方面，TNC 增加了 U251 细胞中每 1000 个细胞的球体数、CD44CD133 细胞百分比和 1/干细胞频率（通过极限稀释分析评估）。在挽救实验中，740 Y-P 降低了 TNC 对 U251 细胞增殖、凋亡、紫杉醇化学敏感性和干性的影响。TNC 通过调节 PI3K/AKT 信号通路在胶质瘤中促进癌细胞增殖和干性，同时抑制凋亡和紫杉醇化学敏感性，从而发挥致癌因子的作用。
