肉桂醛对PC⁃3细胞增殖、EMT及干细胞特性的影响

吴俊檄 唐欲博 令狐熙涛 包广龙 施浩然 文振宇 黄帅 瓦庆德

doi:10.3969/j.issn.1006⁃5725.2023.03.006

实用医学杂志 >

2023 , Vol. 39 >Issue 3: 298 - 308

DOI: https://doi.org/10.3969/j.issn.1006⁃5725.2023.03.006

基础研究

肉桂醛对PC⁃3细胞增殖、EMT及干细胞特性的影响

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1 遵义医科大学第二附属医院骨外科（贵州遵义 563000）；2 中山大学附属第一医院药学部（广州 510260）； 3 广州医科大学附属第二医院骨外科（广州 510260）

网络出版日期: 2023-02-10

基金资助

国家自然科学基金项目（编号：82160577）；广州市科技计划项目（编号：202102010071）；贵州省科技计划项目（编号：黔科

合基础⁃ZK［2021］重点 007）；贵州省优秀青年科技人才培养项目（编号：黔科合平台人才［2021］5613 号）

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Effects of cinnamaldehyde on proliferation，EMT and stem cell characteristics of PC⁃3 cells

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Depart⁃ ment of Orthopedics，the Second Affiliated Hospital of Zunyi Medical University，Zunyi 563000，China

Online published: 2023-02-10

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摘要

目的探索肉桂醛（cinnamaldehyde，CA）对 PC⁃3 细胞增殖、上皮⁃间质转化（epithelial⁃mesen⁃ chymal transition，EMT）及干细胞特性的作用及机制，为 CA 作为潜在抗前列腺癌骨转移药物提供前期实验参考。方法通过 CCK⁃8 法、Transwell 实验、基质黏附实验及成球实验检测 CA 对 PC⁃3 细胞增殖、迁移、侵袭、黏附及成球的影响，qRT⁃PCR 分析 CA 处理 PC⁃3 细胞后 miR⁃143 及 AGO2 mRNA 表达情况，Transwell 实验和基质黏附实验检测单独加入 CA、转染 miR⁃143mimic、AGO2⁃siRNA 同时处理 CA 及 AGO2、同时转染 miR ⁃143mimic 及 AGO2 后 PC ⁃3 细胞的侵袭、迁移及黏附情况。Western blot 实验检测 CA 或转染 miR ⁃ 143mimic 后PC⁃3 细胞AGO2、EMT 及干细胞特性相关蛋白的表达情况。结果 CA 呈浓度依赖性抑制PC⁃3 细胞的增殖、迁移、侵袭、黏附及成球能力，与对照组相比差异均有统计学意义（P < 0.01）；qRT⁃PCR 分析发现 CA 可上调 PC⁃3 细胞中 miR⁃143 表达、抑制 AGO2 mRNA 表达；过表达 miR⁃143 及沉默 AGO2 均能抑制 PC⁃3 细胞的迁移、侵袭及黏附，而同时上调 AGO2 及 CA、同时上调 miR⁃143 及 AGO2 后 PC⁃3 细胞迁移、侵袭及黏附的抑制作用并不明显。此外，过表达 miR⁃143 及 CA 均能抑制 PC⁃3 细胞中 AGO2、ZEB1、Vimentin、 N⁃cadherin、fibronectin 及 CD44、Oct⁃4、c⁃Myc 蛋白的表达。结论 CA 可在体外抑制 PC⁃3 细胞的增殖、迁移、侵袭及黏附能力，此外，CA 可抑制 PC⁃3 细胞的 EMT 及干细胞特性，其机制可能与其能上调 miR⁃143/AGO2 轴有关。

关键词：

肉桂醛

; PC?3; miR?143; 上皮?间质转化; 干细胞特性

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吴俊檄唐欲博令狐熙涛包广龙施浩然文振宇黄帅瓦庆德 .

肉桂醛对PC⁃3细胞增殖、EMT及干细胞特性的影响

[J]. 实用医学杂志, 2023 , 39(3) : 298 -308 . DOI: 10.3969/j.issn.1006⁃5725.2023.03.006

Abstract

Objective To explore the effects of cinnamaldehyde（CA）on the proliferation，epithelial ⁃ mesenchymal transition（EMT）and stem cell characteristics of PC⁃3 cells and their mechanisms，so as to provide reference for preliminary experiment of CA as a potential drug preventing bone metastasis in prostate cancer. Methods The effects of CA on the proliferation，migration，invasion，adhesion and spheres formation of PC⁃3 cells were detected using CCK8，Transwell，matrix adhesion and spheres formation assays. The expressions of miR⁃143 and AGO2 mRNA in PC⁃3 cells after CA treatment were determined by qRT⁃PCR，while the effects on the invasion， migration and adhesion of PC⁃3 cells in the single CA，single transfection of miR⁃143mimic or AGO2⁃siRNA， simultaneous treatment of CA and AGO2，or miR ⁃143 mimic and AGO2 group were detected by Transwell and matrix adhesion assays. Furthermore，the protein expressions of CA or transfection of miR⁃143 mimic on EMT and stem cell characteristics of PC⁃3 cells were determined using western blot. Results CA inhibited the proliferation， migration，invasion，adhesion and spheroid of PC⁃3 cells in a concentration⁃dependent manner，showing statistically significant differences while compared with that in the control group（P < 0.01）. qRT⁃PCR showed that CA up⁃reg⁃ ulated the expression of miR⁃143 and down⁃regulate the expression of AGO2 mRNA in PC⁃3 cells. Overexpression of miR⁃143 or down⁃regulation of AGO2 inhibited the migration，invasion and adhesion of PC⁃3 cells. Also，the inhibition on the invasion，migration and adhesion of PC⁃3 cells was not obvious after the simultaneous up⁃regulation of AGO2 and CA or miR⁃143 and AGO2. In addition，CA and overexpression of miR⁃143 inhibited the expressions of AGO2，ZEB1，vimentin，N⁃cadherin，fibronectin，CD44，Oct⁃4 and c⁃Myc proteins in PC⁃3 cells. Conclusion CA inhibits the proliferation，migration，invasion and adhesion of PC⁃3 cells in vitro；it also inhibits the EMT and stem cell characteristics of PC⁃3 cells，and its mechanism may be related to its ability to up⁃regulate the miR⁃143/ AGO2 axis.

Key words：

cinnamaldehyde

; PC?3; miR?143; EMT; stem cell characteristics ;

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