MiR-650通过靶向结合磷酸烯醇丙酮酸羧激酶1促进口腔鳞状细胞癌细胞增殖和糖酵解  

MiR-650 promotes proliferation and glycolysis of oral squamous cell carcinoma cells by targeting phosphoenolpyruvate carboxykinase 1

作　　者：郭谊 韩炎 周丽芝[3] 王俊华 马跃 鲁大鹏[5] Guo Yi;Han Yan;Zhou Lizhi;Wang Junhua;Ma Yue;Lu Dapeng(Department of Stomatology,Tangshan Hospital of Traditional Chinese Medicine,Tangshan 063011;Department of Stomatology,Wen’an Hospital,Langfang 063000;Department of Stomatology,Tangshan Eighth Hospital,Tangshan 063000;Department of Cardiology,Wen’an Hospital,Langfang 063000;Integrated Emergency Treatment Center,Beijing Stomatological Hospital,Capital Medical University,Beijing 100050,China)

机构地区：[1]唐山市中医医院口腔科,唐山063011 [2]文安县医院口腔科,廊坊063000 [3]唐山市第八医院口腔科,唐山063000 [4]文安县医院医院心内科,廊坊063000 [5]首都医科大学附属北京口腔医院急诊综合治疗中心,北京100050

出　　处：《解剖学杂志》2024年第3期217-224,248,共9页Chinese Journal of Anatomy

基　　金：廊坊市科技支撑计划项目(2018013150)。

摘　　要：目的:分析miR-650是否靶向磷酸烯醇丙酮酸羧激酶1(PCK1)促进口腔鳞状细胞癌(OSCC)细胞增殖和糖酵解。方法:生物信息学筛选出差异表达且与细胞增殖、糖酵解有关的基因。构建稳定过表达PCK1的CAL-27和Tca8113细胞系,分成Vector组和PCK1组。构建稳定过表达miR-650的OSCC细胞,分成miR-NC组、miR-650组和miR-650+PCK1组。CCK-8和克隆形成检测各组细胞增殖能力。裸鼠成瘤实验检测瘤体质量、体积。免疫组织化学检测肿瘤组织中PCK1表达水平。免疫印迹测定各组细胞PCK1蛋白表达水平。葡萄糖和乳酸试剂盒检测各组细胞内葡萄糖和乳酸变化。数据库预测PCK1靶标,双荧光酶素报告实验进行验证。RT-qPCR测定转染后OSCC细胞中miR-650的表达。结果:PCK1在OSCC中表达下调,且与细胞增殖和糖酵解相关。与Vector组相比,PCK1组中PCK1蛋白水平升高;过表达PCK1抑制细胞增殖;过表达PCK1抑制裸鼠移植瘤体积和质量增长;过表达PCK1促进细胞葡萄糖产生和抑制其消耗,同时抑制乳酸产生。与癌旁组织相比,miR-650在OSCC中表达上调。数据库预测及双荧光酶素报告实验证实miR-650与PCK1的靶向关系。与miR-NC组相比,miR-650组中PCK1 mRNA和蛋白表达水平明显降低。miR-650通过调节PCK1表达,促进OSCC细胞增殖,调控细胞糖酵解。结论:miR-650可以通过靶向结合PCK1促进OSCC细胞增殖和糖酵解。Objective:To analyze the mechanism of mi R-650 on promoting proliferation and glycolysis of oral squamous cell carcinoma(OSCC) cells by targeting phosphoenolpyruvate carboxykinase 1(PCK1).Methods:The genes with differential expression and those related to cells proliferation and glycolysis were screened out by bioinformatics.CAL-27 and Tca8113 cell lines with stable overexpression PCK1 were constructed,and they were divided into Vector group and PCK1 group.OSCC cells with stable overexpression mi R-650 were constructed,and they were divided into mi R-NC group,mi R-650 group and mi R-650+PCK1 group.The cells proliferation ability in each group was detected by cell counting kit 8(CCK-8) and colony formation.The weight and volume of tumors were measured by tumor formation assay in nude mice.The expression level of PCK1 in tumor tissues was detected by immunohistochemistry.The expression level of PCK1 protein in each group was detected by Western blotting.The changes of glucose and lactic acid in cells were detected by kits of glucose and lactic acid.The database predicted PCK1 as the target and the dual-luciferase reporter experiment was performed to validate.The mi R-650 in OSCC cells after transfection was detected by real-time fluorescence quantitative polymerase chain reaction(RT-q PCR).Results:PCK1 was down-regulated in OSCC and was related to cells proliferation and glycolysis.Compared with the Vector group,the level of PCK1 protein was significantly increased in the PCK1 group.Overexpression of PCK1 could inhibit cells proliferation,inhibit growth of tumor volume and weight in nude mice.Overexpression of PCK1 could promote the production of glucose and inhibit its consumption,as well as inhibit the production of lactic acid.Mi R-650 was upregulated in OSCC compared with paracancerous tissues.Database prediction and dual-luciferase reporter assays confirmed the targeting relationship between mi R-650 and PCK1.Compared with the mi R-NC group,the expression levels of PCK1 m RNA and protein were significantly dec

关 键 词：口腔鳞状细胞癌 磷酸烯醇丙酮酸羧激酶1 增殖 糖酵解 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]