摘要
目的探究miR-769-5p在皮肤黑素瘤(malignant melanoma,MM)中的表达,以及其对人黑素瘤A375、G361细胞增殖的影响,以期为疾病的临床治疗提供新思路。方法收集本院保存的62例MM患者肿瘤组织以及对应癌旁组织进行研究,实时定量PCR(qRT-PCR)检测组织中miR-769-5p的表达,分析miR-769-5p表达与患者临床病理参数的关系,体外培养A375、G361细胞,转染miR-769-5p inhibitor分为空白对照组、阴性转染组、miR-769-5p转染组,MTT法检测细胞增殖情况;平板细胞克隆实验检测菌落形成情况;流式细胞仪检测细胞凋亡以及周期分布情况;裸鼠模型肿瘤形成实验评估体内肿瘤生长情况。结果 MM组患者miR-769-5p mRNA表达明显高于癌旁组织(P<0.05)。miR-769-5p表达与性别、年龄、病灶溃疡情况无关,与淋巴结转移、临床分期相关(P<0.05)。与HEM-L细胞系相比,A375、G361细胞系miR-769-5p mRNA表达均升高(P<0.05)。与空白对照组、阴性转染组相比,miR-769-5p转染组细胞中miR-769-5p表达、菌落数量、小鼠肿瘤体积均降低,细胞抑制率、G1期DNA含量均升高(P<0.05)。结论 miR-769-5p在MM中呈高表达,抑制其表达后能够影响细胞周期的进展,使细胞阻滞于G0/G1期,进而降低细胞增殖进程。
Objective To explore the expression of miR-769-5p in the skin melanoma(malignant melanoma,MM)and its effects on the proliferation of human melanoma A375 and G361 cells,so as to provide new ideas for the clinical treatment.Methods MM tumor tissues and corresponding paracancerous tissues were collected from the 62 cases,and real time quantitative PCR(qRT-PCR)was used to detect the expression of miR-769-5p in tissues.The relationship between miR-769-5p expression and clinicopathological parameters was analyzed,and A375 and G361 cells were cultured in vitro.The transfected miR-769-5p inhibitor was divided into blank control group,negative transfection group and miR-769-5p transfection group.MTT assay was used to detect cell proliferation.The colony formation was detected by the flat cell cloning experiment;cell apoptosis and cell cycle distribution were detected by flow cytometry;tumor formation experiment in nude mice model was used to evaluate tumor growth in vivo.Results The expression of miR-769-5p mRNA in group MM was significantly higher than that in adjacent tissues(P<0.05).The expression of miR-769-5p was not related to sex,age and lesion ulcers,it was correlated with lymph node metastasis and clinical stage(P<0.05).Compared with HEM-L cell lines,the expression of miR-769-5p mRNA in A375 and G361 cell lines increased(P<0.05).Compared with the blank control group and the negative transfection group,the expression of miR-769-5p,the number of colonies and the tumor volume of the mice in the miR-769-5p transfected group were all decreased,and the cell inhibition rate and the G1 phase DNA content were all increased(P<0.05).Conclusion The inhibition of MiR-769-5pits expression that highly expressed in MM can affect cell cycle progression,block the cells in the G0/G1 phase,furthermore,reduce the process of cell proliferation.
作者
潘南楠
谢治
王琛
李丽丽
李玉秋
黄琦涛
PAN Nannan;XIE Zhi;WANG Chen;LI Lili;LI Yuqiu;HUANG Qitao(Department of Dermatology and Venereology,People's Hospital of Guangxi Zhuang Autonomous Region,Nanning 530021,China)
出处
《中国皮肤性病学杂志》
CAS
CSCD
北大核心
2020年第3期250-256,共7页
The Chinese Journal of Dermatovenereology
基金
广西壮族自治区卫生和计划生育委员会自筹经费科研课题(Z20170319)。
