Paclitaxel Induce Apoptosis of Giant Cells Tumor of Bone via TP53INP1 Signaling
Paclitaxel Induce Apoptosis of Giant Cells Tumor of Bone via TP53INP1 Signaling
复制标题
DOI：
10.1111/os.12414
复制
发表时间：
2019-02-01
影响因子：
2.1
通讯作者：
Lao, Li-Feng
中科院分区：
文献类型：
--
作者：
Xiao, Wei-Yuan;Zong, Zhen;Lao, Li-Feng
目的：为评价紫杉醇的抗肿瘤作用并探讨其分子机制，首先采用CCK-8和凋亡检测法检测紫杉醇对HS 737.T细胞的存活和凋亡作用。接下来，使用RNA-seq和生物信息学来确定差异表达的基因并分析相关途径。实时定量聚合酶链反应用于验证部分差异表达基因（DEG）的准确性。使用CGRGO对差异表达基因的功能分组GO术语进行解码和可视化，并绘制DEG蛋白质-蛋白质相互作用（PPI）网络。Western blotting检测靶基因的表达、Caspase-3和PARP 1的裂解以及p53的磷酸化水平。最后，利用转录组学、生物信息学和RNA干扰技术研究紫杉醇对GCTB细胞的抗肿瘤作用，并探讨其作用机制。结果：紫杉醇对HS 737. T细胞的存活率和诱导凋亡作用具有明显的时间依赖性。RNA-seq和生物信息学分析表明，紫杉醇治疗GCTB与细胞凋亡、死亡受体信号通路、TNF信号通路、TP 53调控的细胞死亡基因转录通路密切相关。Western blot结果显示紫杉醇诱导HS 737.T细胞Caspase-3和PARP 1的裂解，并增加p53的磷酸化水平。RNAi结果显示TP 53 INP 1在HS 737. T细胞中的表达水平明显降低（降低幅度超过70%）。此外，我们发现紫杉醇对TP 53 INP 1缺陷的HS737.T细胞的抑制率低于空载体的HS737.T细胞（分别为19.88%和40.60%）。结论：紫杉醇可通过激活Caspase-3、PARP 1、p53和TP 53 INP 1，抑制HS 737.T细胞增殖，诱导其凋亡。紫杉醇可能是治疗GCTB的有效药物。
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.