circ0091581靶向miR-136对皮肤鳞状细胞癌细胞生物学行为的影响

Effects of circ0091581 on the biological behavior of cutaneous squamous cell carcinoma cells by targeting microRNA-136

目的 circ_0091581是否通过miR-136参与皮肤鳞状细胞癌(CSCC)的发生过程尚不清楚。文章旨在探讨circ_0091581对CSCC细胞生物学行为的影响及其可能作用机制。方法 实时荧光定量聚合酶链反应(qRT-PCR)法检测CSCC组织与癌旁组织中circ_0091581、miR-136的表达量;体外培养人CSCC细胞SCL-1,实验分组:si-NC组、si-circ_0091581组、miR-NC组、miR-136组、si-circ_0091581+anti-miR-NC组、si-circ_0091581+anti-miR-136组;甲基噻唑基四唑(MTT)法、平板克隆形成实验、Transwell实验与流式细胞术分别检测细胞增殖、克隆形成、迁移及凋亡;双荧光素酶报告实验检测circ_0091581与miR-136的靶向关系。结果 circ_0091581在CSCC组织中的表达量(2.45±0.37)较癌旁组织(0.93±0.16)增多(P<0.05),miR-136在CSCC组织中表达量(0.43±0.06)较癌旁组织(0.99±0.11)降低(P<0.05)。与转染si-NC比较,转染si-circ_0091581后细胞克隆形成、迁移能力减低(P<0.01),细胞增殖抑制、凋亡能力增强(P<0.01);与转染miR-NC比较,转染miR-136 mimics后细胞克隆形成、迁移能力减低(P<0.01),细胞增殖抑制、凋亡能力增强(P<0.01);circ_0091581可靶向调控miR-136的表达;共转染si-circ_0091581、anti-miR-136可减弱转染si-circ_0091581对细胞生物学行为的作用。结论 干扰circ_0091581表达可通过靶向调控miR-136表达而减弱CSCC细胞增殖、克隆形成及迁移能力,并可诱导细胞凋亡。

Objective Whether circ_0091581 is involved in the pathogenesis of cutaneous squamous cell carcinoma(CSCC) through miR-136 remains unclear. This paper aims to investigate the effects of circ_0091581 on the biological behavior of cutaneous squamous cell carcinoma and its possible mechanism. Methods Quantitative Real-time PCR(qRT-PCR) was used to detect the expression of circ_0091581 and microRNA-136 in cutaneous squamous cell carcinoma tissues and adjacent tissues. The human cSCC cell lines SCL-1 were cultured in vitro and divided into 6 groups: si-NC group, si-circ_0091581 group, miR-NC group, miR-136 group, si-circ_0091581+anti-miR-NC group, si-circ_0091581+ anti-miR-136 group. Methylthiazolyl tetrazolium(MTT), plate colony formation, transwell and flow cytometry were used to detect cell proliferation, colony formation, migration and apoptosis. The dual luciferase reporter experiment was used to detect the targeting relationship between circ_0091581 and miR-136. Results Compared with adjacent tissues, the expression of circ_0091581 in cutaneous squamous cell carcinoma tissue was increased [(0.93±0.16) vs(2.45±0.37), t=23.548, P<0.01], and the expression of miR-136 was decreased [(0.99±0.11) vs(0.43±0.06), t=27.911, P<0.01]. Compared with transfection of si-NC, the colony formation and migration ability of cells were decreased after transfection with si-circ_0091581(P<0.01), and the cell proliferation inhibition and apoptosis ability were enhanced(P<0.01) Compared with transfection of miR NC, the ability of cell clone formation and migration were decreased after miR-136 mimics were transfected(P<0.01), and the cell proliferation inhibition and apoptosis ability were enhanced. The circ_0091581 could target and regulate the expression of miR-136. Co-transfection of si-circ_0091581 and anti-miR-136 could attenuate the effects of transfection of si-circ_0091581 on the biological behavior of cells. Conclusion The interference of the expression of circ_0091581 could reduce the proliferation, colony formation and migration of the cells of cutaneous squamous cell carcinoma by targeting the expression of miR-136, and induce cell apoptosis.