摘要
目的探究miR-101-3p对口腔鳞状细胞癌(OSCC)细胞体外增殖、凋亡、迁移和侵袭以及体内肿瘤生长的影响,并分析其机制。方法CAL27细胞转染mimics NC、miR-101-3p mimics、inhibitor NC和miR-101-3p inhibitor,RT-qPCR验证转染效率。CCK-8检测细胞增殖能力;流式细胞术检测细胞凋亡水平;Transwell检测细胞迁移和侵袭能力;Western blot检测细胞中APP/MAPK和BMP2/PI3K/AKT信号通路表达。裸鼠皮下注射成瘤后,分为agomiR-NC组、agomiR-101-3p组、antagomiR-NC组和antagomiR-101-3p组,检测移植瘤体积和质量,免疫组化检测移植瘤中Ki-67表达,TUNEL实验检测移植瘤细胞凋亡水平。双荧光素酶报告基因实验验证miR-101-3p与APP和BMP2结合。结果过表达miR-101-3p抑制CAL27细胞增殖、迁移、侵袭和体内肿瘤生长并促进细胞凋亡,降低CAL27细胞中APP、p-ERK1/2、p-JNK3、p-p38、BMP2和p-AKT蛋白表达。抑制miR-101-3p促进CAL27细胞增殖、迁移、侵袭和体内肿瘤生长并抑制细胞凋亡,增加细胞中APP、p-ERK1/2、p-JNK3、p-p38、BMP2和p-AKT蛋白表达。miR-101-3p靶向结合APP和BMP2 mRNA。结论miR-101-3p抑制CAL27细胞增殖、迁移、侵袭和体内肿瘤生长并促进细胞凋亡,其机制与靶向抑制APP介导的MAPK信号通路以及BMP2介导的PI3K/AKT信号通路有关。
Objective To explore the effects of miR-101-3p on the proliferation,apoptosis,migration and invasion of oral squamous cell carcinoma(OSCC)cells in vitro and the tumor growth in vivo,and analyze its mechanism.Methods CAL27 cells were transfected with mimics NC,miR-101-3p mimics,inhibitor NC and miR-101-3p inhibitor,and the transfection efficiency was verified by RT-qPCR.CCK-8 was used to detect cell proliferation.Flow cytometry was used to detect apoptosis of cells.Transwell was used to detect migration and invasion ability of cells.Western blot was used to detect the expressions of APP/MAPK and BMP2/PI3K/AKT signal pathway in cells.After subcutaneous injection,nude mice were divided into agomiR-NC group,agomiR-101-3p group,antagomiR NC group and antagonir 1013P group.The volume and quality of transplanted tumor were detected,the expression of Ki 67 in transplanted tumor was detected by immunohistochemistry,and the apoptosis level of transplanted tumor cells was detected by TUNEL experiment Double luciferase reporter gene experiment verified that miR-101-3p combined with APP and BMP2.Results Overexpression of miR-101-3p inhibited the proliferation,migration,invasion and tumor growth in vivo of CAL27 cells,promoted apoptosis of cells,reduced the expressions of APP,p-ERK1/2,p-JNK3,p-p38,BMP22-2 and p-AKT proteins in CAL27 cells.Inhibiting miR-101-3p promoted the proliferation,migration,invasion of CAL27 cells and tumor growth in vivo,inhibited cell apoptosis,and increased the expressions of APP,p-ERK1/2,p JNK3,p-p38,BMP2 and p-AKT in cells.miR-101-3p targets and binds to APP and BMP2 mRNA.Conclusion MiR-101-3p inhibits the proliferation,migration,invasion and tumor growth of CAL27 cells in vivo,and promotes apoptosis.The mechanism is related to the targeted inhibition of MAPK signaling pathway mediated by APP and PI3K/AKT signaling pathway mediated by BMP2.
作者
刘钟月
杨坤
朱向宇
范新昊
LIU Zhong-yue;YANG Kun;ZHU Xiang-yu;FAN Xin-hao(Department of Stomatology,Kailuan General Hospital,Tangshan O63001;Department of Stomatology,Affiliated Hospital of North China University of Science and Technology,Tangshan O63099,China)
出处
《解剖科学进展》
2025年第5期670-674,共5页
Progress of Anatomical Sciences
基金
河北省医学科学研究重点课题计划项目(20231858)。
