桑黄酮G通过调控PI3K/AKT/mTOR通路抑制胃癌细胞的生长、迁移和侵袭
Kuwanon G inhibits growth,migration and invasion of gastric cancer cells by regulating the PI3K/AKT/mTOR pathway
耿志军 1杨晶晶 2牛民主 3刘馨悦 4施金冉 4刘亦珂 4姚新宇 4张雨路 4张小凤 1胡建国5
作者信息
- 1. 蚌埠医科大学第一附属医院中心实验室,安徽 蚌埠 233000;炎症相关性疾病基础与转化研究安徽省重点实验室,安徽 蚌埠 233004
- 2. 蚌埠医科大学第一附属医院胃肠外科,安徽 蚌埠 233000
- 3. 蚌埠医科大学第一附属医院蚌埠医科大学检验医学院免疫学教研室,安徽 蚌埠 233000
- 4. 炎症相关性疾病基础与转化研究安徽省重点实验室,安徽 蚌埠 233004
- 5. 炎症相关性疾病基础与转化研究安徽省重点实验室,安徽 蚌埠 233004;蚌埠医科大学第一附属医院检验科,安徽 蚌埠 233004
- 折叠
摘要
目的 探讨桑黄酮G(KG)对胃癌细胞的增殖、凋亡、迁移和侵袭的作用和分子机制.方法 通过CCK-8实验、细胞克隆形成实验、裸鼠背部成瘤和Ki-67免疫组织化学染色评估不同浓度KG对胃癌细胞增殖与生长的影响;应用Annexin V-FITC/PI细胞凋亡检测试剂盒、Western blotting分析凋亡相关蛋白表达和Tunel染色评估KG对胃癌细胞凋亡的影响;使用Transwell迁移和侵袭实验与Western blotting分析基质金属蛋白酶表达检测KG对胃癌细胞迁移和侵袭的作用;利用免疫印迹技术和rescue实验验证PI3K/AKT/mTOR通路在KG调控胃癌细胞的增殖、迁移和侵袭的作用.结果 KG以浓度依赖性抑制胃癌细胞的增殖和减少细胞形成克隆数(P<0.05);KG上调凋亡细胞比例,促进cleaved caspase-3、Bcl2-Bax表达并下调Bcl2水平(P<0.05);KG干扰胃癌细胞的迁移和侵袭,并且抑制基质金属蛋白酶2(MMP2)和基质金属蛋白酶9(MMP9)的表达(P<0.05).机制验证显示,KG抑制PI3K/AKT/mTOR通路的激活,且PI3K/AKT/mTOR通路的激活剂IGF-1逆转了KG对胃癌细胞增殖、迁移和侵袭的影响(P<0.05).结论 KG至少部分是通过抑制PI3K/AKT/mTOR通路的激活影响胃癌细胞的增殖、凋亡、迁移和侵袭.
Abstract
Objective To investigate the effects of kuwanon G(KG)on proliferation,apoptosis,migration and invasion of gastric cancer cells and the molecular mechanisms.Methods The effects of KG on proliferation and growth of gastric cancer cells were assessed with CCK-8 assay and cell clone formation assay,by observing tumor formation on the back of nude mice and using immunohistochemical analysis of Ki-67.The effect of KG on cell apoptosis was analyzed using Annexin V-FITC/PI apoptosis detection kit,Western blotting and TUNEL staining.The effects of KG on cell migration and invasion were detected using Transwell migration and invasion assay and Western blotting for matrix metalloproteinase(MMP).The role of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway in KG-mediated regulation of gastric cancer cell proliferation,migration,and invasion was verified by Western blotting and rescue assay.Results KG significantly inhibited proliferation and reduced clone formation ability of gastric cancer cells in a concentration-dependent manner(P<0.05).KG treatment also increased apoptosis,enhanced the expressions of cleaved caspase-3 and Bax,down-regulated Bcl-2,lowered migration and invasion capacities and inhibited the expression of MMP2 and MMP9 in gastric cancer cells(P<0.05).Mechanistic validation showed that KG inhibited the activation of the PI3K/AKT/mTOR pathway,and IGF-1,an activator of the PI3K/AKT/mTOR pathway,reversed the effects of KG on proliferation,migration and invasion of gastric cancer cells(P<0.05).Conclusion KG inhibits proliferation,migration and invasion and promotes apoptosis of gastric cancer cells at least in part by inhibiting the activation of the PI3K/AKT/mTOR pathway.
关键词
胃癌/桑黄酮G/恶性行为/PI3K/AKT/mTOR通路Key words
gastric cancer/kuwanon G/malignant behaviors/PI3K/AKT/mTOR pathway引用本文复制引用
基金项目
安徽省高校协同创新项目(GXXT-2020-020)
安徽省高等学校自然科研究项目(KJ2020A0563)
安徽省卫生健康科研项目(AHWJ2022b088)
出版年
2024
国家科技期刊平台
NSTL
万方数据