| Objective: Paclitaxel is an effective anti-tumor medicine,which has been widely used in various kind of cancers.However,the anti-proliferation and apoptosis mechanism of paclitaxel on giant cell tumors of bone(GCTB)still remains unclear.Hence,the aim of the study is 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 HS737.T cells under treatment of paclitaxel.Next,RNA-seq and bioinformatics were used to determine the differentially expressed genes(DEGs)and to analyze the pathway involved.Real-time Quantitative polymerase chain reaction(RTqPCR)was used to verify the accuracy of some DEGs.ClueGO was used to decode and visualize functionally grouped GO terms of DEGs,and to map the DEGs protein–protein interactions(PPI)network.Western blotting(WB)was used to check the DEGs,the cleavage of Caspase-3 and PARP1,and the phosphorylation level of p53.Finally,transcriptomics,bioinformatics,and RNA interference(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 HS737.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.WB results revealed that paclitaxel induced cleavage of Caspase-3 and PARP1,and increased the phosphorylation level of p53 in HS737.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 HS737.T cells through activating TP53INP1 and p53,improving expression of Caspase10,TRAIL.Paclitaxel may be an effective drug in the management of GCTB. |
