摘要
目的观察microRNA-125b(miR-125b)和信号传导转录激活因子3(STAT3)在皮肤鳞状细胞癌(CSCC)中的表达,并探讨miR-125b靶向STAT3进而调控CSCC发生发展的分子机制。方法采用qRT-PCR法和Western Blot检测32例CSCC组织及其周围正常皮肤组织中以及CSCC细胞系(A431、SCC13和SCL-1)和正常皮肤细胞系HaCaT中miR-125b和STAT3的表达。荧光素酶报告基因实验被用于验证miR-125b对STAT3的靶向作用。采用MTT法和流式细胞术检测miR-125b mimic或STAT3 shRNA对CSCC细胞系(A431、SCC13和SCL-1)细胞增殖、细胞周期和细胞凋亡的影响。采用克隆形成实验、细胞转移和侵袭实验观察miR-125b mimic对CSCC细胞系A431细胞的增殖、转移和侵袭能力的影响。结果与正常皮肤组织和细胞相比,CSCC组织和细胞中miR-125b表达显著下降,而STAT3的表达则显著上调。miR-125b靶向STAT3的3'-UTR发挥对其表达的调控作用。miR-125b mimic或STAT3 shRNA转染后,A431、SCC13和SCL-1细胞的增殖明显受抑制,G0/G1期细胞比例明显增加,并且促进了细胞凋亡。miR-125b mimic能明显抑制A431细胞的克隆形成、细胞转移和侵袭的能力。结论 miR-125b/STAT3的表达异常通过影响细胞的增殖、凋亡和侵袭参与了CSCC的发生发展。二者可成为CSCC新的诊断和治疗靶点。
Objective To investigate the expression of microRNA-125 b(miR-125 b) and signal transduction activator 3(STAT3) in cutaneous squamous cell carcinoma(CSCC),and to explore the molecular mechanism of miR-125 b targeting STAT3 to regulate the occurrence and development of CSCC.Methods qRT-PCR and Western blot were used to detect the expression of microRNA-125 b and STAT3 in 32 cases of CSCC tissues and adjacent normal skin tissues,as well as in CSCC cell lines(A431,SCC13 and SCL-1) and normal skin cell lines HaCaT.Luciferase reporter gene experiment was used to verify that miR-125 b targets STAT3.tetrazolium-based colorimetric assay(MTT) and flow cytometry were used to detect the effects of microRNA-125 b mimic or STAT3 shRNA on cell proliferation,cell cycle and apoptosis of CSCC cell lines(A431,SCC13 and SCL-1).The effects of microRNA-125 b mimic on the proliferation,metastasis and invasion of CSCC cell line A431 were observed by colony formation and Transwell assay.Results Compared with normal skin tissues and cells,the expression of miR-125 b in CSCC tissues and cells decreased significantly,while the expression of STAT3 increased significantly.miR-125 b targets 3 ’-UTR of STAT3 to regulate its expression.After miR-125 b mimic or STAT3 shRNA transfection,the proliferation and cell cycle of A431,SCC13 and SCL-1 cells was significantly inhibited,while the proportion of cells in G0/G1 phase was significantly increased,and cell apoptosis was promoted.Moreover,miR-125 b mimic could significantly inhibit the ability of colony formation,cell migration and invasion in A431 cells.Conclusion The abnormal expression of miR-125 b/STAT3 contributes to the development and progression of CSCC by influencing cell proliferation,apoptosis and invasion.Both of them can be used as new diagnostic and therapeutic targets for CSCC.
作者
付乐凡
田珂
苗国英
冯爱平
FU Lefan;TIAN Ke;MIAO Guoying;FENG Aiping(Union Hospital Affiliated to Huazhong University of science and Technology,Wuhan,Hubei 430022,China;Hebei University of Engineering Affiliated Hospital,Handan,Hebei 056000,China)
出处
《中国热带医学》
CAS
2020年第11期1082-1087,共6页
China Tropical Medicine
