华蟾酥毒基抑制人骨肉瘤细胞迁移和侵袭的机制研究
Mechanism of cinobufagin inhibiting migration and invasion in human osteosarcoma cells
- 上海中医药杂志 2021年55卷第12期 页码：88-93
- 作者机构：
1.新乡医学院第一附属医院肿瘤科（河南 新乡 453100）
2.平顶山市第一人民医院骨科（河南 平顶山 467000）
3.上海中医药大学附属岳阳中西医结合医院肿瘤科（上海 200437）
- 作者简介：
王颖，女，硕士，主治医师，主要从事骨肉瘤侵袭转移机制研究工作
曹飞，主任医师，硕士研究生导师；E-mail：cf771225@163.com
- 基金信息：国家自然科学基金项目(82074268;81503414;81874392;82074231);河南省医学科技攻关项目(SB201903014;LHGJ20191263)
DOI：10.16305/j.1007-1334.2021.2104029
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王颖, 唐晓男, 康小红, 等. 华蟾酥毒基抑制人骨肉瘤细胞迁移和侵袭的机制研究[J]. 上海中医药杂志, 2021,55(12):88-93.
Ying WANG, Xiaonan TANG, Xiaohong KANG, et al. Mechanism of cinobufagin inhibiting migration and invasion in human osteosarcoma cells[J]. Shanghai Journal of Traditional Chinese Medicine, 2021,55(12):88-93.
王颖, 唐晓男, 康小红, 等. 华蟾酥毒基抑制人骨肉瘤细胞迁移和侵袭的机制研究[J]. 上海中医药杂志, 2021,55(12):88-93. DOI： 10.16305/j.1007-1334.2021.2104029.
Ying WANG, Xiaonan TANG, Xiaohong KANG, et al. Mechanism of cinobufagin inhibiting migration and invasion in human osteosarcoma cells[J]. Shanghai Journal of Traditional Chinese Medicine, 2021,55(12):88-93. DOI： 10.16305/j.1007-1334.2021.2104029.
摘要
目的,2,观察华蟾酥毒基对人骨肉瘤（HOS）细胞迁移与侵袭的影响，并探讨其可能的作用机制。,方法,2,应用慢病毒载体LV-vector、LV-over/PLOD2转染HOS细胞并建立稳定转染株HOS/NC、HOS/PLOD2；划痕和细胞侵袭实验检测华蟾酥毒基对肿瘤细胞迁移和侵袭能力的影响；实时定量聚合酶链式反应（qRT-PCR）法检测华蟾酥毒基对,PLOD,2 mRNA表达的影响；Western blot法检测华蟾酥毒基对PLOD2及FAK-JAK/STAT3信号通路相关蛋白（PLOD2、FAK、p-FAK、JAK、p-JAK、STAT3、p-STAT3）表达的影响。,结果,2,划痕和细胞侵袭实验结果显示，华蟾酥毒基可降低HOS细胞的迁移和侵袭能力，与对照组相比，差异有统计学意义（,P,<,0.01），但过表达PLOD2后，华蟾酥毒基抑制HOS/PLOD2细胞迁移和侵袭的能力减弱，与HOS/NC细胞相比，差异有统计学意义（,P,<,0.01）；qRT-PCR结果显示，不同浓度的华蟾酥毒基均可下调HOS细胞,PLOD,2 mRNA表达水平；Western blot结果显示，华蟾酥毒基可下调HOS/NC细胞中PLOD2及FAK-JAK/STAT3通路相关蛋白p-FAK、p-JAK、p-STAT3等的表达，但对HOS/PLOD2细胞中PLOD2及FAK-JAK/STAT3信号通路相关蛋白的作用明显减弱。,结论,2,华蟾酥毒基抑制骨肉瘤迁移与侵袭的机制可能与下调PLOD2，进而阻断FAK-JAK/STAT3信号通路有关。
Abstract
Objective,2,To investigate the effects of cinobufagin on human osteosarcoma (HOS) cells migration and invasion, and explore its potential mechanism.,Methods,2,HOS cells were transfected with LV-vector and LV-over/PLOD2 to establish stable transfected cell lines HOS/NC and HOS/PLOD2. Scratch test and Transwell invasion assays were applied to detect the effects of cinobufagin on the migration and invasion ability of osteosarcoma cells. Quantitative real-time polymerase chain reactions(qRT-PCR) was adopted to detect the effects of cinobufagin on ,PLOD,2 mRNA expression. Western blot assays were used to detect the effect of cinobufagin on the expressions of PLOD2 and FAK-JAK/STAT3 signal pathway related proteins(PLOD2, FAK, p-FAK, JAK, p-JAK, STAT3, p-STAT3).,Results,2,The results of scratch test and Transwell invasion assays showed that cinobufagin could reduce the migration and invasion ability of HOS cells, and compared with the control group the difference was statistically significant(,P,<,0.01). However, overexpression of PLOD2 significantly attenuated cinobufagin-induced suppression of migration and invasion in HOS cells, and the difference was statistically significant compared with HOS/NC cells(,P,<,0.01). The result of qRT-PCR showed that the expressions of ,PLOD,2 mRNA were significantly down-regulated in HOS cells which were treated by different doses cinobufagin. The Western blot analysis showed that cinobufagin significantly down-regulated the expressions of PLOD2 and FAK-JAK/STAT3 signal pathway related proteins p-FAK, p-JAK and p-STAT3 in HOS/NC cells, but the effect on those proteins in HOS/PLOD2 cells was significantly weakened.,Conclusion,2,The mechanism of cinobufagin inhibiting the invasion and migration of osteosarcoma may be related to the down-regulation of PLOD2, which in turn blocks the FAK-JAK/STAT3 signal pathway.
关键词
骨肉瘤华蟾酥毒基侵袭迁移赖氨酸羟化酶2中药研究
Keywords
osteosarcomacinobufagininvasionmigrationPLOD2traditional Chinese herbal medicine research
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