lncRNA UCA1靶向调控p21对黑色素瘤细胞增殖、侵袭的影响  

Long Noncoding RNA UCA1 Regulates Proliferation and Invasion of Melanoma Cells through Targeting p21

作　　者：闫璐[1] 邱贤文[1] 王穗海[2] 魏姗姗 黄东兰 谭锐[1] 李鹏飞 关志广 Yan Lu;Qiu Xianwen;Wang Suihai(Department of Dermatology,Zhujiang Hospital of Southern Medical University,Guangzhou 510280,China2.College of Biotechnology,Southern Medical University,Guangzhou 510280,China;College of Biotechnology,Southern Medical University,Guangzhou 510280,China)

机构地区：[1]南方医科大学珠江医院皮肤科,广东510280 [2]南方医科大学生物技术学院,广东510280 [3]广州医科大学附属肿瘤医院放疗科,广东510280 [4]广东省台山市人民医院烧伤整形外科,广东529200

出　　处：《华中科技大学学报（医学版）》2019年第2期162-167,182,共7页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：2018年度南方医科大学"科研启动计划"基础研究前期启动项目(No.QD2018N021)

摘　　要：目的探讨长链非编码RNA尿路上皮癌相关1(UCA1)在黑色素瘤组织和细胞中的表达及其介导的肿瘤细胞增殖和侵袭行为。方法采用实时荧光定量PCR检测UCA1在黑色素瘤组织和3种黑色素瘤细胞株(M21、B16、A375)中的表达情况;利用siRNA技术敲低M21、A375细胞UCA1的表达,MTT法检测细胞增殖能力,Transwell迁移和侵袭实验检测细胞迁移和侵袭能力,流式细胞术检测细胞周期分布和凋亡率;Real-time PCR和Western blot检测敲低M21、A375细胞UCA1表达后p21表达变化情况。结果 UCA1在黑色素瘤组织和黑色素瘤细胞株(M21、B16、A375)中表达明显高于正常皮肤色素痣组织和正常黑色素细胞HEM,差异具有统计学意义(P<0.05,P<0.01)。将UCA1 siRNA(si-UCA1)分别转染至黑色素瘤细胞M21和A375,与对照组(si-NC)比较,si-UCA1组增殖活性明显降低,G_0/G_1期细胞比例明显升高,细胞凋亡率明显增加,差异具有统计学意义(P<0.05,P<0.01)。M21和A375细胞的si-UCA1组穿膜细胞数和迁移细胞数明显低于si-NC组(均P<0.01)。p21 mRNA在黑色素瘤组织表达明显低于正常皮肤色素痣组织,差异具有统计学意义(P<0.01);相关分析显示,黑色素瘤组织UCA1与p21 mRNA表达呈显著负相关(r=-0.291,P<0.01)。敲低M21和A375细胞中UCA1表达明显升高p21 mRNA和蛋白表达水平,而转染p21 siRNA则显著降低由si-UCA1诱导的p21表达(均P<0.01)。结论长链非编码RNA UCA1在黑色素瘤中高表达,UCA1可能通过靶向调控p21参与黑色素瘤细胞的增殖和侵袭。Objective To determine expression of long noncoding RNA UCA1 on melanoma tissues and cells and its effect on the cell proliferation and invasion. Methods The expression of UCA1 in melanoma tissues and cells(M21,B16,A375)was detected by real-time PCR,and siRNA transfection was carried out to knockdown the expression of UCA1 in M21 and A375 cells.The effect of the knockdown of UCA1 on cell proliferation was detected by MTT assay,and cytometry analysis was conducted to investigate cell cycle and apoptosis.Migration and invasion of melanoma cells were detected by Transwell assays.Furthermore,Western blotting was used to check the protein expression of p21 after inhibition of UCA1 expression in M21 and A375 cells. Results The expression level of UCA1 was significantly higher in melanoma tissues and cells(M21,B16,A375)than in non-tumor tissues and human epidermal melanocytes(P<0.05,P<0.01).MTT assay showed that cell proliferation was noteworthily decreased in M21 and A375 cells transfected with si-UCA1.Meanwhile,down-regulation of UCA1 boosted the number of cells in G 0/G1 phase and increased number of apoptosis cells in M21 and A375 cells(P<0.05,P<0.01).Furthermore,low expression of UCA1 reduced the number of migratory and invasive cells in M21 and A375 cells compared to the negative control(P<0.01).Expression of p21 mRNA was more highly expressed in the normal skin tissues than in tumor tissues(P<0.01).Correlation analysis displayed that UCA1 was negatively correlated with p21 expression in melanoma tissues(r=-0.291,P<0.01).Moreover,knockdown of UCA1 increased both mRNA and protein level of p21,which was then decreased again by the knockdown of p21. Conclusion Long noncoding RNA UCA1 is highly expressed in melanoma tissues and cells.UCA1 may be involved in proliferation and invasion of melanoma cells through targeted regulating p21.

关 键 词：长链非编码RNA 尿路上皮癌相关1 黑色素瘤 增殖 侵袭 

分 类 号：R739.5[医药卫生—肿瘤]