ABSTRACT
OBJECTIVE: To evaluate the antitumor capability and to investigate the underlying molecular mechanism of paclitaxel. METHODS: First, cck-8 and apoptosis assays were used to determine survival and apoptotic effects of HS 737.T cells under treatment of paclitaxel. Next, RNA-seq and bioinformatics were used to determine the differentially expressed genes and to analyze the pathway involved. Quantitative real-time polymerase chain reaction was used to verify the accuracy of some differentially expressed genes (DEG). ClueGO was used to decode and visualize functionally grouped GO terms of differentially expressed genes, and to map the DEG protein-protein interactions (PPI) network. Western blotting was used to check the expression of target genes, the cleavage of Caspase-3 and PARP1, and the phosphorylation level of p53. Finally, transcriptomics, bioinformatics, and RNAi were used to estimate the antitumor capability and to identify the underlying mechanisms of paclitaxel in GCTB. RESULTS: Our data revealed that paclitaxel had significant time-dependent effects on the viability and induced apoptosis of HS 737.T cells. RNA-seq and bioinformatics analysis showed that apoptosis, death receptor signaling pathway, TNF signaling pathway, and TP53 regulated transcription of cell death genes pathway were closely associated with paclitaxel in the treatment of GCTB. Western bolt results revealed that paclitaxel induced cleavage of Caspase-3 and PARP1, and increased the phosphorylation level of p53 in HS 737.T cells. RNAi results showed that the expression level of TP53INP1 was significantly decreased in HS737.T cells (the decrease was more than 70%). In addition, we found that the inhibitory ratios of paclitaxel on HS737.T cells deficient in TP53INP1 were less than in HS737.T cells with empty vector (19.88 and 40.60%, respectively). Hence, our data revealed that TP53INP1 regulated paclitaxel-driven apoptosis in HS737.T cells. CONCLUSION: Paclitaxel can significantly repress cell proliferation and induce apoptosis of HS 737.T cells through activating Caspase-3, PARP1, p53, and TP53INP1. Paclitaxel may be an effective drug in the management of GCTB.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Apoptosis/drug effects , Bone Neoplasms/pathology , Carrier Proteins/physiology , Giant Cell Tumor of Bone/pathology , Heat-Shock Proteins/physiology , Paclitaxel/pharmacology , Bone Neoplasms/metabolism , Caspase 3/metabolism , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Survival/drug effects , Computational Biology/methods , Drug Evaluation, Preclinical/methods , Gene Expression Regulation, Neoplastic/drug effects , Giant Cell Tumor of Bone/metabolism , Humans , Poly (ADP-Ribose) Polymerase-1/metabolism , Signal Transduction/drug effects , Tumor Cells, Cultured , Tumor Suppressor Protein p53/metabolismABSTRACT
STUDY DESIGN: A retrospective study of clinical cases. PURPOSE: To evaluate the efficacy of continuous irrigation and drainage for early postoperative deep wound infection after posterior instrumented spinal fusion. SUMMARY OF BACKGROUND DATA: Aggressive debridement and irrigation has been recommended to treat postoperative wound infections after instrumented spinal fusion. However, this method of management, indicating repeating visits to the operating room until the wound is clean enough for closure, often results in prolonged hospitalization, increased cost, and sometimes compromise of the desired outcome. We hypothesize that repeat visits to the operating room for debridements can be avoided by aggressive debridements and primary closure with continuous irrigation and drainage for postoperative wound infections. METHODS: From 2004 to 2009, 23 patients with early postoperative deep wound infections after spinal fusion with instrumentation were surgically treated with thorough debridement and primary closure with continuous irrigation and drainage. All patients were followed up for 30.6 months (range, 24-54 mo). RESULTS: The mean duration of irrigation was 12.0 days (range, 7-16 d). In 21 patients (91.3%), the wound healed after continuous irrigation. The removal of the instrumentation or cages was not required in any case. Spinal fusion was achieved in all cases, except 1, where the patient developed a pseudoarthrosis at the L4-L5 level after L4-S1 fusion. The mean ODI for these 23 patients improved significantly from 53.4±18.7 preoperatively to 18.3±11.2 at the final follow-up visit (P<0.001). The mean JOA scores increased significantly from 15.5±4.1 preoperatively to 24.3±3.8 at the final follow-up (P<0.001). CONCLUSIONS: Continuous irrigation and drainage is an effective and safe method for the treatment of early postoperative deep wound infection after posterior instrumented spinal fusion.
