lncRNA COLCA1/miR-423-5p/FAM172A分子轴对皮肤鳞状细胞癌A431细胞的增殖和迁移的影响

目的探讨长链非编码RNA(long-chain non-coding RNA, lncRNA)COLCA1在皮肤鳞状细胞癌(cutaneous squamous cell carcinoma, CSCC)组织中的表达及其对细胞增殖和迁移的影响、分子机制。方法采用qRT-PCR法检测COLCA1在正常皮肤组织和CSCC组织、细胞系及人角质形成细胞系中的表达;选择表达最低的细胞系,分别转染空白质粒(对照组)或COLCA1过表达质粒(实验组);采用MTT法和细胞划痕实验分别检测过表达COLCA1对CSCC细胞系增殖和迁移的影响;通过生物信息学法预测COLCA1潜在的分子机制。采用qRT-PCR和Western blot法检测过表达COLCA1对靶基因表达的影响。结果 CSCC组织中COLCA1的表达显著低于正常皮肤组织(1.01±0.20 vs 5.71±0.78,P<0.01)。CSCC细胞系中COLCA1的表达显著低于人角质形成细胞系(P<0.01),A431细胞系中的表达最低(P<0.01)。与对照组相比,过表达质粒可有效促进COLCA1的表达(1.00±0.04 vs 9.31±0.99,P<0.01)。过表达COLCA1可抑制A431细胞的增殖能力和迁移能力(P均<0.01)。COLCA1可能与miR-423-5p互补结合,miR-423-5p可能与FAM172A mRNA互补结合。与对照组相比,过表达COLCA1可抑制A431细胞中miR-423-5p的表达(1.00±0.05 vs0.20±0.05,P<0.01),并促进FAM172A在mRNA和蛋白的表达(P<0.01)。结论 COLCA1在CSCC组织和细胞系中呈低表达,过表达COLCA1可能通过miR-423-5p/FAM172A分子轴抑制CSCC细胞系A431的增殖和迁移能力。

Effect of lncRNA COLCA1/miR-423-5p/FAM172A molecular axis on the proliferation and migration of cutaneous squamous cell carcinoma A431 cells

Purpose To explore the expression of long-chain non-coding RNA(lncRNA)COLCA1 in cutaneous squamous cell carcinoma(CSCC)tissues,its effect on cell proliferation and migration,and its molecular mechanism.Methods qRT-PCR was used to detect the expression of COLCA1 in CSCC tissue and normal skin tissue,CSCC cell lines and human keratinocyte cell line.The cell line with the lowest expression was selected and the blank plasmid(control group)or COLCA1 overexpression plasmid(experiment group)transfected respectively.The effects of overexpression of COLCA1 on the proliferation and migration of CSCC cell lines were detected by MTT method and cell scratch test,respectively.The potential molecular mechanism of COLCA1 was predicted through bioinformatics.The effect of overexpression of COLCA1 on target gene expression was detected by qRT-PCR and Western blot.Results The expression of COLCA1 in CSCC tissue was significantly lower than that in normal skin tissue(1.01±0.20 vs 5.71±0.78,P<0.01).The expression of COLCA1 in CSCC cell line was significantly lower than that of human keratinocyte cell line(P<0.01),and the expression in A431 cell line was the lowest(P<0.01).Compared with control group,overexpression plasmid could effectively promote the expression of COLCA1(1.00±0.04 vs 9.31±0.99,P<0.01).Overexpression of COLCA1 could inhibit the proliferation ability(P<0.05)and migration ability(P<0.01)of A431 cells.COLCA1 might be complementary to miR-423-5p,and miR-423-5p might be complementary to FAM172A mRNA.Compared with control group,overexpression of COLCA1 could inhibit the expression of miR-423-5p in A431 cells(1.00±0.05 vs 0.20±0.05,P<0.01),and promote the expression of FAM172A at the mRNA and protein levels(P<0.01).Conclusion COLCA1 is underexpressed in CSCC tissues and cell lines.Overexpression of COLCA1 might inhibit the proliferation and migration of CSCC cell line A431 through the miR-423-5p/FAM172A molecular axis.