摘要
目的探讨微小RNA(miRNA)-21抑制剂联合基于石墨烯量子点负载顺铂纳米药物系统(GPt)对黑色素瘤细胞凋亡、迁移和侵袭的影响及其机制。方法培养黑色素瘤B16F10细胞,分为对照组、纳米药24 h组、纳米药48 h组、纳米药72 h组,利用四甲基偶氮唑蓝(MTT)筛选出合适的给药剂量和用药时间;然后再分为miRNA-21抑制剂组、纳米药组、miRNA-21抑制剂+纳米药组。采用Transwell实验检测细胞侵袭能力,划痕实验检测细胞迁移能力,流式细胞仪检测细胞周期和凋亡情况,聚合酶链反应(PCR)和蛋白质印迹法(Western blot)检测miRNA-21、B细胞淋巴瘤/白血病-2(Bcl-2)、p53和Bcl-2相关X蛋白(BAX)的表达情况。再通过皮下注射B16F10细胞制备裸鼠黑色素瘤模型,检测各组肿瘤重量。结果筛选出用药作用时间与剂量分别为72 h和5.0μl,miRNA-21抑制剂具有明显的干扰效果,可用于后期实验。与对照组相比,纳米药组和miRNA-21抑制剂组细胞的侵袭能力、迁移能力均下降,G1期细胞比例升高,S期细胞比例下降,凋亡增多,两者联用后效果更明显。miRNA-21抑制剂组和纳米药组的Bcl-2蛋白和mRNA相对表达量均下降,两者联用后下降更加明显,p53、BAX蛋白和mRNA相对表达量均升高,两者联用后升高更加明显。与对照组相比,其他各组小鼠肿瘤重量均下降,并且miRNA-21抑制剂+纳米药组下降最明显。对照组的肿瘤组织中血管分布丰富,肿瘤细胞排列紧密,肿瘤细胞生长活跃,大小相对一致,其他各组肿瘤细胞排列疏松,细胞生长不活跃。结论GPt联合miRNA-21抑制剂能够诱导黑色素瘤细胞发生凋亡,并能抑制细胞的迁移和侵袭能力。
Objective To investigate the effect of microRNA-21(miRNA-21)inhibitor combined with graphene quantum dots loaded cisplatin nano-drug system(GPt)on the apoptosis,migration and invasion of melanoma cells and its mechanism.Method B16F10 melanoma cells were cultured and divided into control group,nano-drug 24 h group,nanodrug 48 h group and nano-drug 72 h group.Methyl thiazolyl tetrazolium(MTT)assay was used to screen the appropriate dose and administration time of nano-drug;then they were divided into miRNA-21 inhibitor group,nano-drug group,miRNA-21 inhibitor+nano-drug group.Transwell assay was used to detect cell invasion ability,scratch test was used to detect cell migration ability,flow cytometry was used to detect cell cycle and apoptosis.Polymerase chain reaction(PCR)and Western blot were used to detect the expression of miRNA-21,B cell lymphoma/leukemia-2(Bcl-2),p53 and Bcl-2 associated X protein(BAX).And the B16F10 cells were subcutaneously injected into nude mice to establish a melanoma model,and the tumor weight of each group were detected.Result The screened time and dose of nano-drug were 72 h and 5.0μl,respectively,miRNA-21 inhibitor had obvious interference effect,which could be used for later experiments.Compared with the control group,the invasion and migration ability of the cells in the nano-drug group and the miRNA-21 inhibitor group were decreased,the proportion of cells in G1 phase was increased,the proportion of cells in S phase was decreased,and the proportion of apoptosis cells was increased.The effect of the combination of the two groups was more obvious.The relative expression of Bcl-2 protein and mRNA in the miRNA-21 inhibitor group and the nano-drug group decreased,and the decrease was more obvious after the combination of the two groups,the expression of p53 and BAX protein and mRNA increased,and the increase was more obvious after the combination of the two groups.Compared with the control group,the tumor weight of the other groups decreased,and the miRNA-21 inhibitor+nano-drug group d
作者
游小龙
周正宇
陈雯
魏小勇
傅爱荣
袁分钱
李敏
凌琦
YOU Xiaolong;ZHOU Zhengyu;CHEN Wen;WEI Xiaoyong;FU Airong;YUAN Fenqian;LI Min;LING Qi(Department of Head and Neck Oncology Surgery,Jiangxi Provincial Cancer Hospital,Nanchang 330029,Jiangxi,China)
出处
《癌症进展》
2024年第5期501-506,共6页
Oncology Progress
基金
江西省卫生健康委科技计划项目(20203545)。
