摘要
目的探讨长链非编码RNA(lncRNA)FOXD3-AS1靶向miR-128-3p/PDK1对黑色素瘤细胞增殖和侵袭的调控作用。方法转染组-1,转染组-2,转染组-3分别转染pcDNA-FOXD3-AS1、pcDNA-FOXD3-AS1与miR-128-3p mimics的阴性对照(mimic NC)、pcDNA-FOXD3-AS1与miR-128-3p mimics,以未进行转染的细胞作为对照组。用实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测黑色素瘤细胞和正常人表皮黑色素细胞FOXD3-AS1和miR-128-3p的表达差异;用双荧光素酶报告基因实验验证FOXD3-AS1与miR-128-3p的相互作用;用细胞计数试剂盒-8(CCK-8)法检测细胞的增殖能力;用Transwell小室法检测细胞的侵袭;用蛋白质印迹法检测3-磷酸肌醇依赖性蛋白激酶1(PDK1)蛋白的表达水平。结果对照组、转染组-1,转染组-2,转染组-3细胞24 h的增殖率分别为(100.00±0.00)%,(168.03±15.71)%,(163.58±6.24)%,(124.62±8.93)%;侵袭的细胞数分别为(21.33±2.08),(38.33±4.89),(40.00±2.65),(25.33±2.52)个;PDK1的表达水平分别为0.36±0.02,1.19±0.08,1.24±0.11,0.48±0.03。转染组-1与对照组比较,转染组-3与转染组-2比较,差异均有统计学意义(P<0.05)。结论FOXD3-AS1通过靶向miR-128-3p/PDK1轴,促进黑色素瘤细胞的增殖与侵袭。
Objective To investigate the regulatory effects of long non-coding RNA(lncRNA)FOXD3-AS1 targeting miR-128-3p/PDK1 on the proliferation and invasion of melanoma cells.Methods Transfection-1 group,transfection-2 group,transfection-3 group were transfected with PcDNA-FOXD3-AS1,negative control(mimic NC)of miR-128-3p mimics and pcDNA-FOXD3-AS1,pcDNA-FOXD3-AS1 and miR-128-3p mimics,cells without transfection were as control group.Differential expression of FOXD3-AS1 and miR-128-3p in melanoma cells and normal human epidermal melanocytes was detected by fluorescence quantitative polymerase chain reaction(qRT-PCR);the interaction between FOXD3-AS1 and miR-128-3p was verified by dual-luciferase reporter gene assay;the proliferation ability of the cells was detected by cell counting kit-8(CCK-8)assay;the invasion of the cells was detected by Transwell assay;and the expression of 3-phosphoinositide-dependent protein kinase 1(PDK1)protein was detected by Western blot.Results The proliferation rates of control group,transfection-1 group,transfection-2 group and transfection-3 group cells at the 24 h were(100.00±0.00)%,(168.03±15.71)%,(163.58±6.24)%and(124.62±8.93)%,respectively;the number of invaded cells were(21.33±2.08),(38.33±4.89),(40.00±2.65)and(25.33±2.52),respectively;and the expression of PDK1 were 0.36±0.02,1.19±0.08,1.24±0.11 and 0.48±0.03.There were significant differences between transfection-1 group and control group,and between transfection-3 group and transfection-2 group(P<0.05).Conclusion FOXD3-AS1 promotes the proliferation and invasion of melanoma cells by targeting the miR-128-3p/PDK1 axis.
作者
巩伟亮
徐媛媛
董文丽
张本利
严旭
郭洪飞
马学良
史文平
李文娟
GONG Wei-liang;XU Yuan-yuan;DONG Wen-li;ZHANG Ben-li;YAN Xu;GUO Hong-fei;MA Xue-liang;SHI Wen-ping;LI Wen-juan(Department of Dermatology,Tengzhou Central People’s Hospital,Tengzhou 277599,Shandong Province,China;Department of Dermatology,Shandong Provincial Hospital,Jinan 250021,Shandong Province,China)
出处
《中国临床药理学杂志》
CAS
CSCD
北大核心
2021年第16期2167-2169,2183,共4页
The Chinese Journal of Clinical Pharmacology
