FIS1通过P53通路抑制骨肉瘤对顺铂化疗的耐药
| 作者: |
1谭瑞,
1段伟,
2王子君,
3,4袁鹏,
1常乐,
1甘璐,
5权鹏鹤,
6史喜德,
1李沫,
1叶正旭
1 空军军医大学第一附属医院骨科，西安 710032 2 空军军医大学基础医学院学员一大队一队，西安 710032 3 空军军医大学第二附属医院疼痛科，西安 710038 4 空军军医大学基础医学院生理与病理生理教研室，西安 710032 5 河北省军区石家庄第二离职干部休养所，石家庄 050000 6 中国人民解放军32356部队卫生队，青海 格尔木 816000 |
| 通讯: |
李沫
Email: limo0729@hotmail.com 叶正旭 Email: yzengx@fmmu.edu.cn |
| DOI: | 10.3978/j.issn.2095-6959.2021.10.002 |
| 基金: | 国家重点研发计划(2017YFB1104104)；陕西省自然科学基金(2019SF-054)。 |
摘要
目的：探讨线粒体分裂蛋白1(mitochondrial fission protein 1，FIS1)在顺铂(cisplatin，CDDP)耐药的骨肉瘤(osteosarcoma，OS)组织中的表达及对CDDP耐药OS细胞凋亡的作用。方法：采用免疫组织化学法(immunohistochemistry，IHC)和蛋白质印迹法检测CDDP耐药的OS组织(CDDP-resistance组)和CDDP敏感的OS组织(CDDP-sensity组)中FIS1的表达水平，生物信息学方法分析FIS1在OS中的相关作用通路，流式细胞术检测CDDP耐药细胞凋亡及过表达FIS1对细胞活性氧(reactive oxygen species，ROS)的作用，蛋白质印迹法检测FIS1对P53信号通路的作用，OS公共数据分析FIS1与P53的相关性。结果：FIS1在18例CDDP-sensity组中表达明显比其在15例CDDP-resistance组中表达增高；FIS1在OS耐CDDP细胞系MG63/Cis-R细胞中的蛋白质表达低于普通MG63组；人OS组织中与FIS1相关的前10位KEGG (Kyoto Encyclopedia of Genes and Genomes)通路，凋亡通路排在首位。在OS耐CDDP细胞系MG63/Cis-R细胞中，FIS1高表达组细胞与对照组相比，细胞凋亡比例增高(P<0.05)，ROS水平降低(P<0.05)，P53、BAX和cleaved Caspase3蛋白质表达上调，BCL-2分子表达降低；在OS组织中同样发现FIS1与P53、BAX、cleaved Caspase3表达正相关，与BCL-2表达负相关。结论：FIS1通过激活P53通路进而调控CDDP耐药OS细胞的凋亡，而FIS1可能在OS CDDP耐药中发挥重要作用，有望成为OS化疗耐药的新靶点。
关键词：
骨肉瘤；FIS1；顺铂耐药；细胞凋亡；P53
Mitochondrial fission protein 1 inhibits cisplatin resistance in osteosarcoma through P53 pathway
CorrespondingAuthor: LI Mo Email: limo0729@hotmail.com
DOI: 10.3978/j.issn.2095-6959.2021.10.002
Foundation: This work was supported by the National Key Research and Development Program (2017YFB1104104), and Natural Science Foundation of Shaanxi Province (2019SF-054), China.
Abstract
Objective: To explore the expression of mitochondrial fission protein 1 (FIS1) in the tissue of cisplatin (CDDP)-resistant osteosarcoma (OS) and its effect on apoptosis of CDDP-resistant OS cells. Methods: Immunohistochemistry (IHC) and Western blot were used to detect FIS1 expression in CDDP resistant and sensitive OS tissues. Bioinformatics analyse the relative pathways of FIS1 in OS. The apoptosis and reactive oxygen species (ROS) of CDDP resistant cells are detected by flow cytometry. The effect of FIS1 on the P53 signaling pathway was detected by Western blot. The correlation between FIS1 and P53 was analyzed by OS public data. Results: FIS1 expression in 18 CDDP-sensitive OS tissues was significantly higher than that in 15 CDDP-resistant OS tissues. FIS1 protein expression in OS CDDP resistant cell line MG63/CIS-R was lower than that in normal MG63 group. Among the top 10 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways associated with FIS1 in human OS, apoptotic pathways were at the top 1. In OS CDDP resistant cell line MG63/Cis-R, FIS1 high expression group showed an increased apoptosis rate and a decreased ROS compared with the control group (P<0.05); the expression of P53, BAX and cleaved Caspase3 proteins were up-regulated, while the expression of BCL-2 molecules was decreased. There was a significant positive correlation between FIS1 and P53, BAX, cleaved Caspase3 expression in OS tissues, but negatively correlated with BCL-2 expression. Conclusion: FIS1 regulates apoptosis of CDDP-resistant OS cells by activating P53 pathway, and FIS1 may play an important role in CDDP-resistant OS and is expected to be a new target for chemotherapy resistance of OS.
Keywords：
osteosarcoma; FIS1; cisplatin resistance; apoptosis; P53