lncRNA DNAJC3-AS1靶向miR-4319调控皮肤鳞状细胞癌增殖、迁移和侵袭

lncRNA DNAJC3-AS1 regulates the proliferation,migration and invasion of cutaneous squamous cell carcinoma cells by targeting miR-4319

目的研究长链非编码RNA(lncRNA)DNAJC3-AS1靶向miR-4319对皮肤鳞状细胞癌(CSCC)细胞增殖、迁移和侵袭的影响。方法RT-qPCR检测39例CSCC患者癌组织及癌旁组织中DNAJC3-AS1和miR-4319表达。将CSCC细胞A431分为si-NC组、si-DNAJC3-AS1组、miR-NC组、miR-4319组、pcDNA组、pcDNA-DNAJC3-AS1组、si-DNAJC3-AS1+anti-miR-NC组、si-DNAJC3-AS1+anti-miR-4319组。CCK-8法、平板克隆实验、划痕愈合实验、Transwell实验分别用于检测A431细胞活力、克隆形成、迁移和侵袭能力。双荧光素报告实验分析DNAJC3-AS1和miR-4319的靶向关系。结果CSCC组织中DNAJC3-AS1表达上调,miR-4319表达下调。与si-NC组比较,si-DNAJC3-AS1组A431细胞活力、克隆形成数、划痕愈合率、侵袭细胞数显著降低(P<0.05),miR-4319表达水平显著升高(P<0.05)。与miR-NC组比较,miR-4319组A431细胞活力、克隆形成数、划痕愈合率、侵袭细胞数显著降低(P<0.05)。DNAJC3-AS1与miR-4319直接特异性结合。pcDNA-DNAJC3-AS1组A431细胞miR-4319表达水平显著低于pcDNA组(P<0.05)。与si-DNAJC3-AS1+anti-miR-NC组比较,si-DNAJC3-AS1+anti-miR-4319组A431细胞活力、克隆形成数、划痕愈合率、侵袭细胞数显著升高(P<0.05)。结论CSCC中DNAJC3-AS1呈高表达,miR-4319呈低表达。抑制DNAJC3-AS1通过靶向上调miR-4319可降低CSCC细胞增殖、迁移和侵袭。因此,抑制DNAJC3-AS1/miR-4319轴可能成为潜在的CSCC治疗策略。

Objective To study the effects of long non-coding RNA(lncRNA)DNAJC3-AS1 targeting miR-4319 on the proliferation,migration and invasion of cutaneous squamous cell carcinoma(CSCC)cells.Methods RT-qPCR was performed to assess the expression of DNAJC3-AS1 and miR-4319 in cancer tissues of 39 CSCC patients.CSCCA431 cells were divided into si-NC group,si-DNAJC3-AS1 group,miR-NC group,miR-4319 group,pcDNA group,pcDNA-DNAJC3-AS1 group,si-DNAJC3-AS1+anti-miR-NC group,si-DNAJC3-AS1+anti-miR-4319 group.CCK-8 method,plate cloning assay,scratch healing assay,and Transwell assay were used to evaluate cell viability,clone formation,migration and invasion ability of A431,respectively.The targeting relationship between DNAJC3-AS1 and miR-4319 was verified using dual luciferase reporter assay.Results The expression of DNAJC3-AS1 was up-regulated,while the expression of miR-4319 was down-regulated in CSCC tissues.Compared with the si-NC group,the cell viability,clone formation numbers,scratch healing rate and invasion numbers of A431 in si-DNAJC3-AS1 group were significantly reduced(P<0.05),and miR-4319 expression level was significantly increased(P<0.05).Compared with the miR-NC group,the cell viability,clone formation numbers,scratch healing rate and invasion numbers of A431 in miR-4319 group were significantly reduced(P<0.05).DNAJC3-AS1 directly and specifically bound to miR-4319.miR-4319 expression of A431 cells in the pcDNA-DNAJC3-AS1 group was significantly lower than that in the pcDNA group(P<0.05).Compared with the si-DNAJC3-AS1+anti-miR-NC group,the cell viability,clone formation numbers,scratch healing rate and invasion numbers of A431 in the si-DNAJC3-AS1+anti-miR-4319 group were significantly increased(P<0.05).Conclusion DNAJC3-AS1 was highly expressed while miR-4319 was low expressed in CSCC.DNAJC3-AS1 inhibition could reduce the proliferation,migration and invasion ability of CSCC cells by targeting and up-regulating miR-4319.Therefore,inhibition of DNAJC3-AS1/miR-4319 axis might be a potential therapeutic strategy for CSCC.