RESUMO
The abnormal activation of hedgehog (HH) signaling pathway plays an important role in the development and progression of glioblastoma (GBM). As a transcription factor at the end of the HH pathway, the final effector of glioma-associated oncogene homoglog-1 (GLI1) is an important target in the treatment of GBM. The study was designed to evaluate the anti-tumor activities and mechanisms of a novel GLI1 inhibitor FL18 in GBM. MTT and colony formation assay were performed to determine anti-proliferation activity of FL18 in vitro. The effect of FL18 on cell apoptosis was measured by flow cytometry (FCM) analysis. Transwell experiment was used to explore the inhibitory activity of FL18 in cell invasion. In vivo experiments, the subcutaneously transplanted and orthotopic U-87 MG GBM xenograft model were used to study the activity of FL18 on tumor growth. The optimized dual report gene screening model was used to detect the effect of FL18 on the transcriptional activity of GLI1. Western blot assay was used to study the mechanisms of action of FL18. The results showed that the IC50 of FL18 in glioblastoma was in the nanomole level in vitro. It was observed that 22.5 and 45 mg·kg-1 FL18 reduced the tumor volume with the rate of 55.4% and 89.8% in xenograft model in mice in situ. The IC50 of FL18 on the inhibition of GLI1 transcriptional activity was 3.32×10-11 mol·L-1 analyzed by the optimized dual report gene screening model. By the Western blot experiments, it was proved that FL18 inhibited expression of GLI1 without influencing the upstream canonical HH/SMO signaling and cross-talk oncogenic pathway, such as ERK and AKT signaling. The results also demonstrated that FL18 significantly downregulated GLI1 target genes such as Bcl-2, MMP2 and MMP9 and increased the expression of c-caspase3, c-PARP and Bax. These data suggest that FL18 may generate the anti-glioma activity by inhibition of GLI1.
RESUMO
<p><b>OBJECTIVE</b>To study the indication and character of the lateral-cervical approach for treating dumble-shape neurogenic tumors in cervical spine.</p><p><b>METHODS</b>Retrospectively review the clinical data of 14 dumble-shape neurogenic tumors in cervical spine, from October 2005 to October 2011. Among them 8 were males and 6 were females, range from 11 to 60 years old. The maximum tumor diameter range from 3.0 to 8.0 cm, with an average of 4.8 cm; the intraspinal tumor diameter range from 1.3 to 3.8 cm, with an average of 2.1 cm. According to Asazuma classification, 9 cases were type IIc, 2 cases were type IIIb, 2 cases were type IV, 1 case was type VI. Involving the neck segment C(1)-C(2) in 1 case, C(2)-C(3) in 1 case, C(3)-C(4) in 2 cases, C(4)-C(5) in 2 cases, C(5)-C(6) in 3 cases, C(6)-C(7) in 4 cases and C(2)-C(4) in 1 case. All cases performed surgery with general anethesia. The head and neck surgeon performed surgery with lateral cervical approach, in the space between the anterior and the medius scalenus, exposed the transverse process and the intervertebral foramen as the anatomy marker, resected the extraspinal tumor part. The neurosurgery expanded the intervertebral foramen, and resected the intraspinal tumor with microscope, and repaired the dura. Then head and neck surgeon closed the wounds.</p><p><b>RESULTS</b>Pathology proved 3 neurolimmoas and 11 Schwannomas, 12 cases received gross total resection, 2 cases received subtotal resection, the average blood loss during operation was 292 ml, the average operation time was 129 minutes, the average stay in hospital days was 7.1 days. The vertebral artery were exposed in 2 cases, and no vertebral artery injury occurred, there were 3 cases dissect the cervical nerve roots. No cerebrospinal fluid leakage, hematoma, newly branchial plexus injury, sympathic nerve injury or tracheal edema occurred. In 3 to 24 months, with an average of 13.5 months follow-up period, 2 cases with subtotal resection had no tumor progression, and 12 cases with gross total resection had no tumor recurrence.</p><p><b>CONCLUSIONS</b>Lateral-cervical approach is minimal invasive, easily to perform and recovery fine. It can be adopt for Asazuma type IIc, IIIb and IV tumors which not grow over the midline in spine and expand to deep layer of the deep cervical fascia out spine.</p>