实用医学杂志 ›› 2022, Vol. 38 ›› Issue (9): 1076-1081.doi: 10.3969/j.issn.1006⁃5725.2022.09.007

• 基础研究 • 上一篇    下一篇

桦木酸在酸性微环境中对PC⁃3细胞生物学行为及破骨细胞分化的作用

黄帅1 黄国樑1 令狐熙涛1 谢尚延1 瓦庆德2 郭远清3 黄彦1   

  1. 1 广州医科大学附属第二医院骨科(广州 510260);2 遵义医科大学第二附属医院骨科(贵州遵义 563000); 3 中山大学附属第五医院脊柱外科(广州 528406)

  • 出版日期:2022-05-10 发布日期:2022-05-10
  • 通讯作者: 黄彦 E⁃mail:drhy229@163.com
  • 基金资助:
    国家自然科学基金(编号:81872172,81860322);广东省自然科学基金面上项目(编号:2019A1515010690);广州市科技计划项目(编号:202002030014,202102010071);珠海市科技计划项目(编号:20191210E030075);贵州省科技计划项目(编号:黔
    科合基础⁃ZK[2021]重点 007)

Study on the effect of betulinic acid on the biological behavior and osteoclast differentiation of PC⁃3 cells in acidic microenvironment

HUANG Shuai*,HUANG Guoliang,LINGHU Xitao,XIE Shangyan,WA Qingde, GUO Yuanqing,HUANG Yan.   

  1. Department of Orthopedics,the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China 

  • Online:2022-05-10 Published:2022-05-10

摘要:

目的 研究桦木酸在酸性微环境中对 PC⁃3 细胞增殖、迁移、黏附、成球、干细胞特性及破骨 细胞分化的影响。方法 使用 pH 7.4 pH 6.5 的培养基培养 PC⁃3 细胞,然后加入浓度为 0、1、5、10、15 20、30、40 μmol/L 的桦木酸继续培养,24 h 后采用 CCK⁃8 法和 Transwell 检测桦木酸对 PC⁃3 细胞增殖与迁 移的影响,通过黏附试验、成球试验检测 BA PC⁃3 细胞干细胞特性的变化,通过破骨细胞分化实验检 测桦木酸对破骨细胞分化的影响,利用 qPCR Western blot 检测 PC⁃3 细胞中 CD31 CD44 蛋白的表达。 结果 CCK⁃8 结果表明与中性对照组(pH 7.4)相比,酸性微环境(pH 6.5)能促进 PC⁃3 细胞增殖,而桦木酸 呈浓度依赖性抑制 PC⁃3 细胞在酸性微环境中的增殖(P < 0.05)。此外,酸性微环境促进 PC⁃3 细胞的迁 移、黏附、瘤体形成及 CD31 CD44 表达,在此基础上加入桦木酸后,PC⁃3 细胞的迁移能力、黏附、瘤体形成、干细胞特征和破骨细胞分化明显下调(P < 0.05)。结论 桦木酸可抑制酸性微环境中 PC⁃3 细胞的增 殖、迁移、干细胞特性及破骨细胞分化。

关键词:

桦木酸, 前列腺癌骨转移, 酸性微环境, 肿瘤干细胞, 破骨细胞生成

Abstract:

Objective To investigate the effects of Betulinic acid(BA)on proliferation,migration adherence properties,tumorsphere formation,stem cell property and osteoclast differentiation in human prostate can⁃ cer PC⁃3 cells in acidic microenvironment. Methods Culturing PC⁃3 cells in pH 7.4 and pH 6.5 medium. Effects of BA on proliferation and migration of PC⁃3 cells were assessed with the CCK⁃8 assay and Transwell assays,respectively. Effects of BA on stem cell characteristics of PC⁃3 cells were evaluated by cell⁃matrix adhesion assay. The osteoclasto⁃ genesis activity of BA in RAW264.7 cells was examined by osteoclast differentiation assay. The express levels of CD31 and CD44 in PC⁃3 cells were measured by qPCR and Western blot. Results The result of CCK⁃8 test showed that acidic medium significantly enhanced cell viability of PC⁃3 cells than neutral medium,whereas BA suppressed the proliferation of PC⁃3 cells in a concentration⁃dependent under acidic microenvironment(P < 0.05). In addition the acidic medium also significantly enhanced cell migration,adherence,tumor sphere formation and expressed CD31,CD44 of PC ⁃3 cells than neutral medium. After that,BA was added and the migration,adherence,tumor sphere formation,stem cell property and osteoclast differentiation of PC ⁃ 3 cells were significantly inhibited(P < 0.05). Conclusion Betulinic acid significantly inhibited the proliferation,migration,adherence,tumor sphere formation,stem cell property and osteoclast differentiation of PC⁃3 cells under acidic microenvironment.

Key words:

betulinic acid, prostate cancer bone metastasis, tumor acidic microenvironment, tumor stem cells, osteoclastogenesis