LncRNA-DLEU2对恶性黑色素瘤细胞增殖影响  

Effects of lncRNA-DLEU2 on proliferation of malignant melanoma

作　　者：田冬梅[1] 俞梅琴 曾琪[2] TIAN Dong-mei;YU Mei-qin;ZENG Qi(Department of Emergency,First Affiliated Hospital of Anhui Medical University,Hefei 230022,P.R.China;Department of Ultrasound nFirst Affiliated Hospital of Gannan Medical University,Ganzhou 341000,P.R.China)

机构地区：[1]安徽医科大学第一附属医院急诊科,安徽合肥230022 [2]赣南医学院第一附属医院超声科,江西赣州341000

出　　处：《中华肿瘤防治杂志》2020年第22期1799-1803,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：江西省科技厅自然基金(20142BAB215055);江西省卫生厅科技计划(20143124);江西省教育厅(GJJ14696)。

摘　　要：目的基于数据挖掘技术筛选出与恶性黑色素瘤增殖相关的lncRNA分子,并以人恶性黑色素瘤A375细胞为靶细胞,观察验证淋巴细胞白血病缺失基因2(deleted in lymphocytic leukemia 2,DLEU2)的作用。方法利用siRNA技术下调人恶性黑色素瘤A375细胞DLEU2表达,通过real-time PCR验证沉默效果,CCK8和克隆形成实验检测细胞增殖、流式细胞术检测细胞周期。结果DLEU2-siRNA成功敲低A375细胞DLEU2表达。CCK8实验表明,DLEU2-siRNA组的A375细胞增殖能力(0.9292±0.103)较对照组(1.343±0.069)和NC-siRNA组(1.355±0.081)降低,差异有统计学意义,F=47.821,P<0.001。克隆形成实验表明,DLEU2-siRNA组的A375细胞克隆形成数(45.33±14.07)较对照组(184.5±36.04)和NC-siRNA组(197.50±26.65)降低,差异有统计学意义,F=58.076,P<0.001。细胞周期实验检测表明,DLEU2-siRNA组的A375细胞G0/G1期百分比(53.167±5.742)%较对照组(39.667±3.559)%和NC-siRNA组(39.833±3.488)%增多,差异有统计学意义,F=18.143,P<0.001;G2/M期百分比(26.500±6.058)%较对照组(42.000±4.980)%和NC-siRNA组(41.000±5.621)%减少,差异有统计学意义,F=14.549,P<0.001。结论敲低DLEU2表达能够导致恶性黑色素瘤A375细胞增殖受到抑制和细胞周期阻滞,其可能作为潜在的评估恶性黑色素瘤患者预后的标志物和治疗靶标。OBJECTIVE The purpose of this study was to screen the lncRNAs related to the proliferation of malignant melanoma based on data mining technology,and to validate the effect of deleted in lymphocytic leukemia 2(DLEU2)in malignant melanoma A375 cells.METHODS siRNA was used to silence DLEU2 expression in human malignant melanoma A375 cells.Silencing effects were verified by real-time PCR,cell proliferation were detected by CCK8 and clone formation,and cell cycle was measured via flow cytometry.RESULTS DLEU2 expression in A375 cells was successfully knocked down by siRNA.CCK8 assay indicated the proliferation ability in DLEU2-siRNA group(0.9292±0.103)was decreased compared with that in control group(1.343±0.069)or NC-siRNA group(1.355±0.081),and the differences were statistically significant(F=47.821,P<0.001).Clone formation assay indicated the numbers of clone formation in DLEU2-siRNA group(45.33±14.07)were decreased compared with these in control group(184.5±36.04)or NC-siRNA group(197.50±26.65),and the differences were statistically significant(F=58.076,P<0.001).Cell cycle assay showed the percentages of G0/G1 phase cells in DLEU2-siRNA group(53.167±5.742)%were increased compared with these in control group(39.667±3.559)%or NC-siRNA group(39.833±3.488)%,and the differences were statistically significant(F=18.143,P<0.001).Percentages of G2/M phase cells in DLEU2-siRNA group(26.500±6.058)%were decreased compared with these in control group(42.000±4.980)%or NC-siRNA group(41.000±5.621)%,and the differences were statistically significant(F=14.549,P<0.001).CONCLUSION Knockdown of DLEU2 expression could cause inhibition of cell proliferation and cell cycle arrest of A375 cells,and it could be used as a potential target in the therapy and prognosis of patients with malignant melanoma.

关 键 词：淋巴细胞白血病缺失基因2 恶性黑色素瘤 细胞增殖 细胞周期 

分 类 号：R739.5[医药卫生—肿瘤]