首页|桔梗皂苷D对人骨肉瘤细胞活力、迁移、侵袭、凋亡和细胞周期的影响

桔梗皂苷D对人骨肉瘤细胞活力、迁移、侵袭、凋亡和细胞周期的影响

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目的:明确桔梗皂苷D(platycodin D,PD)对骨肉瘤细胞活力、迁移、侵袭、凋亡和细胞周期的影响并探讨可能机制.方法:MG63和U2OS细胞分别分为对照组和PD(6.25、12.5、25、50 和100 µmol/L)处理组,CCK-8法检测细胞活力并筛选处理浓度.MG63分为对照(0 µmol/L)组和PD(15 µmol/L)处理组,U2OS分为对照(0 µmol/L)组和PD(25 µmol/L)处理组,划痕和Transwell实验检测细胞迁移和侵袭能力;Hoechst 33342染色法观察细胞核形态;流式细胞术分析细胞周期和凋亡率;Western blot实验检测cleaved caspase-3、cleaved PARP、c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)、p-JNK、B细胞淋巴瘤蛋白2(B-cell lymphoma-2,Bcl-2)、Bcl-2关联X蛋白(Bcl-2-ssociated X protein,BAX)、基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)、MMP-9、细胞周期蛋白依赖性激酶4(cyclin-dependent kinase 4,CDK4)、细胞周期蛋白D1(cyclin D1)、CDK1、cyclin B1、细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)和p-ERK的蛋白表达水平;选取MG63进行蛋白质组测序分析.结果:与对照组比较,PD处理后,MG63和U2OS细胞的存活率、划痕愈合率和侵袭细胞数显著下降(P<0.05),细胞凋亡率显著升高(P<0.05),且细胞出现核浓染、碎裂等现象;MG63和U2OS的细胞周期分布分别在G2/M期和G0/G1期增多;PD处理组BAX、cleaved caspase-3、cleaved PARP和p-JNK/JNK蛋白表达较对照组升高,Bcl-2、MMP-2、MMP-9、CDK4、cyclin D1、CDK1、cyclin B1和p-ERK/ERK蛋白表达较对照组降低(P<0.05).蛋白质组测序分析显示PD作用与细胞分裂、细胞周期、局部黏附、凋亡及丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)信号通路等有关.结论:PD抑制骨肉瘤细胞体外的活力、迁移、侵袭,并促进凋亡和周期阻滞,其机制可能与调控MAPK信号通路有关.
Effects of platycodin D on osteosarcoma cells in vitro
AIM:To investigate the impact of platycodin D(PD)on the viability,migration,invasion,apop-tosis and cell cycle of osteosarcoma cells in vitro,along with its underlying mechanisms.METHODS:Human osteosarco-ma cells MG63 and U2OS were divided into control group(0 µmol/L)and PD treatment group(6.25,12.5,25,50 and 100 µmol/L,respectively).Human osteosarcoma cells MG63 and U2OS were categorized into control groups(0 µmol/L PD)and PD treatment groups(6.25,12.5,25,50 and 100 µmol/L).The CCK-8 assay determined cell viability and identified effective treatment concentrations.MG63(15 µmol/L PD)and U2OS(25 µmol/L PD)were specifically ana-lyzed.Cell scratch and Transwell assays assessed migration and invasion.Hoechst 33342 staining examined nuclear mor-phological changes.Flow cytometry analyzed cell cycle distribution and apoptosis rate.Western blot measured protein ex-pression levels:cleaved caspase-3,cleaved PARP,c-Jun N-terminal kinase(JNK),p-JNK,B-cell lymphoma-2(Bcl-2),Bcl-2-ssociated X protein(BAX),matrix metalloproteinase 2(MMP-2),MMP-9,cyclin-dependent kinase 4(CDK4),cyclin D1,CDK1,cyclin B1,extracellular signal-regulated kinase(ERK)and p-ERK.Proteome sequencing of MG63 cells was performed.RESULTS:PD treatment significantly decreased cell survival,scratch healing rate,and invasive cell numbers,while increasing apoptosis rates(P<0.05).Morphological changes such as nuclear hyperchroma-tism and fragmentation were observed in PD-treated cells.PD induced G2/M phase arrest in MG63 and G0/G1 phase arrest in U2OS cells.PD treatment upregulated BAX,cleaved caspase-3,cleaved PARP,and p-JNK/JNK,while downregulat-ing Bcl-2,MMP-2,MMP-9,CDK4,cyclin D1,CDK1,cyclin B1,and p-ERK/ERK(P<0.05).Proteome sequencing re-vealed PD's involvement in cell division,cell cycle regulation,focal adhesion,apoptosis,and the MAPK signaling path-way.CONCLUSION:PD inhibits cell viability,migration,and invasion of osteosarcoma cells in vitro,while promoting apoptosis and inducing cell cycle arrest.These effects are likely mediated through modulation of the MAPK signaling path-way.

platycodin Dosteosarcomacell viabilitymigrationinvasionapoptosis

祝欣萍、杨佳璐、高志鹏、王梦肖、常世君、贾迪、赵炜明

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黑龙江中医药大学,黑龙江 哈尔滨 150040

齐齐哈尔医学院,黑龙江 齐齐哈尔 161006

黑龙江中医药大学附属第二医院,黑龙江 哈尔滨 150001

桔梗皂苷D 骨肉瘤 细胞活力 细胞迁移 细胞侵袭 细胞凋亡

黑龙江省博士后面上资助项目

LBH-Z23036

2024

中国病理生理杂志
中国病理生理学会

中国病理生理杂志

CSTPCD北大核心
影响因子:1.065
ISSN:1000-4718
年,卷(期):2024.40(10)