摘要目的 探讨长链非编码RNA(lncRNA)肌动蛋白纤维相关蛋白1?反义RNA1 (AFAP1?AS1)通过靶向微小RNA?545?3p(miR?545?3p)调控口腔鳞状细胞癌增殖、侵袭和转移的机制.方法 收集2017年3月至2018年5月在本院行手术切除的口腔鳞癌患者组织30例,同时收集相应癌旁正常组织,实时荧光定量PCR(qRT?PCR)检测口腔鳞癌组织及癌旁正常组织中AFAP1?AS1和miR?545?3p表达;以口腔鳞癌SCC15细胞为研究对象,荧光素酶报告基因实验检测miR?545?3p对AFAP1?AS1活性的影响;构建AFAP1?AS1抑制表达和miR?545?3p过表达的人口腔鳞癌SCC15细胞, CCK?8实验、划痕实验和Transwell实验分别检测细胞增殖、迁移和侵袭能力.结果 与癌旁正常组织比较,口腔鳞癌组织中AFAP1?AS1表达升高(5.5±1.79比1.00±0.16,P<0.01),miR?545?3p表达降低(0.35±0.07比1.00±0.11,P<0. 05);双荧光素酶报告基因检测结果显示AFAP1?AS1可靶向负调控miR?545?3p表达;敲减AFAP1?AS1能明显抑制SCC15细胞增殖、迁移和侵袭能力,促进miR?545?3p的表达;过表达miR?545?3p后,明显抑制SCC15细胞增殖、迁移和侵袭能力;同时敲减AFAP1?AS1和抑制miR?545?3p表达,SCC15细胞增殖、迁移和侵袭能力明显增强.结论 lncRNA AFAP1?AS1在口腔鳞癌中高表达,通过靶向抑制miR?545?3p可调控口腔鳞癌细胞增殖、迁移和侵袭能力.

abstractsObjective To investigate the mechanism underlying long?chain non?coding RNA (lncRNA)actin filament?associated protein 1?antisense RNA1(AFAP1?AS1)regulation of proliferation, invasion and metastasis of oral squamous cell carcinoma by targeting microRNA?545?3p(miR?545?3p). Methods Thirty patients with oral squamous cell carcinoma,who underwent resection in our hospital between March 2017 and May 2018,were included in the study,and the corresponding normal tissues adjacent to the carcinoma were collected. Real?time fluorescent quantitative PCR(qRT?PCR)was used to determine the expression of AFAP1?AS1 and miR?545?3p in oral squamous cell carcinoma and adjacent normal tissues. The oral squamous cell carcinoma SCC15 cells were included as the subject in the study,and the dual luciferase reporter gene assay was used to examine the effect of miR?545?3p on AFAP1?AS1 activity. The SCC15 cells with suppressed AFAP1?AS1 and overexpressed miR?545?3p. CCK?8 assay,scratch test and Transwell assay were used to determine the cell proliferation,migration and invasion,respectively. Results Compared with the normal tissues adjacent to the carcinoma,the expression level of AFAP1?AS1 increased in the oral squamous cell carcinoma tissues (5.15 ± 1.79 vs. 1.00 ± 0.16,P<0.01),whereas the expression level of miR?545?3p decreased (0.35 ± 0.07 vs. 1.00 ± 0.11,P<0.05). The findings of dual luciferase reporter gene assay showed that AFAP1?AS1 targeted and negatively regulated miR?545?3p expression. Knockdown of AFAP1?AS1 significantly inhibited the proliferation,migration and invasion of SCC15 cells,and promoted the expression of miR?545?3p in SCC15 cells. After overexpression of AFAP1?AS1,the proliferation,migration and invasion of SCC15 cells were significantly inhibited. At the same time, the proliferation,migration and invasion of SCC15 cells were significantly enhanced after knockdown of AFAP1?AS1 and inhibition of miR?545?3p expression. Conclusions LncRNA AFAP1?AS1 is highly expressed in oral squamous cell carcinoma and may inhibit the proliferation,migration and invasion of oral squamous cell carcinoma by targeting inhibition of miR?545?3p.