Objective: To investigate the effects of long-chain non-coding RNA ASB16 antisense RNA1 (ASB16-AS1) on the proliferation, migration and invasion of esophageal cancer cells by targeting microRNA (miR )-1258. Methods: Forty pairs of esophageal cancer tissues and matched adjacent tissues (distance of tumor margin>3 cm) resected in Xinxiang Central Hospital from May 2016 to July 2017 were collected. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expressions of ASB16-AS1 and miR-1258 in esophageal cancer tissues and adjacent tissues. The small interfering RNA negative control (si-NC), ASB16-AS1 small interfering RNA (si-ASB16-AS1), miR-negative control mimics (miR-NC), miR-1258 mimics (miR-1258), si-ASB16-AS1 and anti-miR-NC, si-ASB16-AS1 and anti-miR-1258, si-ASB16-AS1 and anti-miR-1258 were transfected into Eca109 cells, respectively. Methyl thiazolyl tetrazolium (MTT) was utilized to detect the cell viability. Transwell assays were applied to detect cell migration and invasion. Double luciferase reporting experiment and qRT-PCR were used to confirm the relationship between ASB16-AS1 and miR-1258. Results: The expression levels of ASB16-AS1 and miR-1258 in esophageal cancer tissues were 2.95±0.27 and 0.62±0.06, respectively. Compared with 1.00±0.06 and 1.00±0.07 in adjacent tissues, the difference was statistically significant (P<0.05). The cell viability of the si-NC group at 48 h and 72 h were 0.81±0.07 and 1.15±0.11, while those of si-ASB16-AS1 group were 0.46±0.04 and 0.62±0.06 (P<0.05). The numbers of cell migration and invasion in the si-NC group were 86.32±8.24 and 71.29±7.15, respectively, while those of si-ASB16-AS1 group were 43.22±4.31 and 32.36±3.58, respectively, the differences were statistically significant (P<0.05). The cell viability of the miR-NC group at 48 h and 72 h were 0.84±0.08, 1.18±0.12, while those of miR-1258 group were 0.55±0.05, 0.71±0.07 (P<0.05). The migration and invasion numbers of the miR-NC group were (83.15±8.31) and (75.33±7.51), while those of miR-1258 group were (49.58±4.23) and (38.42±3.84), respectively, the differences were statistically significant (P<0.05). The cell viability of the si-ASB16-AS1+ anti-miR-NC group at 48 h and 72 h were 0.45±0.04, 0.61±0.06, while those of si-ASB16-AS1+ anti-miR-1258 group were 0.72±0.07, 0.98±0.08; The migration and invasion numbers of cells in the si-ASB16-AS1+ anti-miR-NC group were 44.36±4.41 and 31.69±3.85, respectively, while those of si-ASB16-AS1+ anti-miR-1258 group were 72.65±7.27 and 61.22±6.14, respectively, and the differences were statistically significant (P<0.05). ASB16-AS1 targeted negative regulation of miR-1258 expression. Conclusions: ASB16-AS1 upregulates in esophageal cancer. ASB16-AS1 promotes the proliferation, migration and invasion of esophageal cancer cells by targeting miR-1258.
目的： 探讨长链非编码RNA ASB16-AS1靶向miR-1258对食管癌细胞增殖、迁移和侵袭的影响。 方法： 收集2016年5月至2017年7月于新乡市中心医院接受手术治疗的40例食管癌患者的食管癌及癌旁正常组织(距离癌组织>3 cm)。采用实时荧光定量聚合酶链反应(qRT-PCR)检测食管癌及癌旁正常组织中ASB16-AS1和miR-1258基因的表达水平。采用脂质体法将si-NC(si-NC组)、si-ASB16-AS1(si-ASB16-AS1组)、miR-NC(miR-NC组)、miR-1258模拟物(miR-1258组)、si-ASB16-AS1+anti-miR-NC(si-ASB16-AS1+anti-miR-NC组)、si-ASB16-AS1+anti-miR-1258(si-ASB16-AS1+anti-miR-1258组)分别转染至Eca109细胞。采用四甲基偶氮唑蓝法检测细胞活力，Transwell实验检测细胞迁移和侵袭能力，双荧光素酶报告实验和qRT-PCR检测ASB16-AS1与miR-1258之间的相互作用。 结果： 食管癌组织中ASB16-AS1、miR-1258的表达水平分别为2.95±0.27和0.62±0.06，与癌旁正常组织(分别为1.00±0.06和1.00±0.07)比较，差异均有统计学意义(均P<0.05)。培养48、72 h后，si-NC组细胞吸光度(A)值分别为0.81±0.07和1.15±0.11，均高于si-ASB16-AS1组[分别为0.46±0.04和0.62±0.06，均P<0.05]。si-NC组细胞的迁移和侵袭数分别为(86.32±8.24)个和(71.29±7.15)个，均高于si-ASB16-AS1组[分别为(43.22±4.31)个和(32.36±3.58)个，均P<0.05]。培养48、72 h后，miR-NC组细胞的A值分别为0.84±0.08和1.18±0.12，均高于miR-1258组(分别为0.55±0.05和0.71±0.07，均P<0.05)；miR-NC组细胞的迁移和侵袭数分别为(83.15±8.31)个和(75.33±7.51)个，均高于miR-1258组[分别为(49.58±4.23)个和(38.42±3.84)个，均P<0.05]；si-ASB16-AS1+anti-miR-NC组细胞的A值分别为0.45±0.04和0.61±0.06，均低于si-ASB16-AS1+anti-miR-1258组(分别为0.72±0.07和0.98±0.08，均P<0.05)；si-ASB16-AS1+anti-miR-NC组细胞的迁移和侵袭数分别为(44.36±4.41)个和(31.69±3.85)个，均低于si-ASB16-AS1+anti-miR-1258组[分别为(72.65±7.27)个和(61.22±6.14)个，均P<0.05]。ASB16-AS1靶向负调控miR-1258的表达。 结论： 食管癌中ASB16-AS1呈高表达，ASB16-AS1通过靶向miR-1258可促进食管癌细胞的增殖、迁移和侵袭。.
Keywords: ASB16-AS1; Cell proliferation; Esophageal neoplasms; MiR-1258; Migration and invasion.