Li Guo-Wei, Jin Yan-Ping, Sheng Min-Feng
School of Rehabilitation Science, Nanjing Normal University of Special Education, Nanjing, Jiangsu, China.
School of Nursing, Jiangsu Health Vocational College, Nanjing, Jiangsu, China.
J Cell Mol Med. 2025 Sep;29(17):e70842. doi: 10.1111/jcmm.70842.
To clarify the regulatory effects of miR-192 on the malignant phenotypes of glioma cells. We used PCR, WB and immunofluorescence to detect regulatory factors in glioma samples. Then, we chose lentiviral plasmid transfection to construct cell models. We used CCK-8 and colony formation to evaluate the proliferation ability of these cells and used Transwell/scratch tests to evaluate their invasion ability. CD133-expressing GSCs were observed under a microscope, and their stemness properties were evaluated. We constructed a tumour-bearing model via subcutaneous inoculation. Tumour growth curves and tumour weights were determined subsequently. The proteins involved in the miR-192-EGR1/HOXB9 loop were evaluated via IHC staining. MiR-192 was significantly reduced in glioma samples, and this factor downregulated EGR1 and HOXB9 via targeted binding, thus forming a semi-open loop. Moreover, the proliferation, invasion and migration of glioma cells overexpressing miR-192 were significantly decreased. These malignant phenotypes were abrogated completely with EGR1 or HOXB9 overexpression. Similarly, these changes were essentially consistent with MT marker expression and the stem-like properties in glioma cells. Meanwhile, miR-192 inhibits the tumorigenesis of glioma cells via the EGR1-HOXB9 loop. MiR-192 could inhibit MT in glioma cells through the EGR1-HOXB9 loop. Thus, it reduces their stemness and abrogates their malignant phenotypes.
为阐明miR-192对胶质瘤细胞恶性表型的调控作用。我们采用聚合酶链反应（PCR）、蛋白质免疫印迹法（WB）和免疫荧光法检测胶质瘤样本中的调控因子。然后，我们选择慢病毒质粒转染来构建细胞模型。我们使用细胞计数试剂盒-8（CCK-8）和集落形成实验来评估这些细胞的增殖能力，并使用Transwell小室实验/划痕实验来评估它们的侵袭能力。在显微镜下观察表达CD133的胶质瘤干细胞（GSCs），并评估其干性特征。我们通过皮下接种构建荷瘤模型。随后测定肿瘤生长曲线和肿瘤重量。通过免疫组化染色评估参与miR-192-早期生长反应因子1（EGR1）/同源盒基因B9（HOXB9）环的蛋白质。miR-192在胶质瘤样本中显著降低，该因子通过靶向结合下调EGR1和HOXB9，从而形成一个半开放环。此外，过表达miR-192的胶质瘤细胞的增殖、侵袭和迁移能力显著降低。随着EGR1或HOXB9的过表达，这些恶性表型完全消除。同样，这些变化与胶质瘤细胞中的微管（MT）标记物表达和干细胞样特征基本一致。同时，miR-192通过EGR1-HOXB9环抑制胶质瘤细胞的肿瘤发生。miR-192可通过EGR1-HOXB9环抑制胶质瘤细胞中的MT。因此，它降低了它们的干性并消除了它们的恶性表型。
