Jia Z, Wang P S, Yang Y, Zhu D Y, Wang Z H, Wang W
The Fourth Clinical College of Xinxiang Medical College, The Second Department of Thoracic Tumor, Xinxiang Central Hospital, Xinxiang 453000, China.
The Fourth Clinical College of Xinxiang Medical College, Surgery Department of Xinxiang Central Hospital, Xinxiang 453000, China.
Zhonghua Zhong Liu Za Zhi. 2021 Jul 23;43(7):762-768. doi: 10.3760/cma.j.cn112152-20200509-00430.
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. 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. 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 (<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 (<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 (<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 (<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 (<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 (<0.05). ASB16-AS1 targeted negative regulation of miR-1258 expression. ASB16-AS1 upregulates in esophageal cancer. ASB16-AS1 promotes the proliferation, migration and invasion of esophageal cancer cells by targeting miR-1258.
通过靶向微小RNA(miR)-1258研究长链非编码RNA ASB16反义RNA1(ASB16-AS1)对食管癌细胞增殖、迁移和侵袭的影响。收集2016年5月至2017年7月在新乡市中心医院切除的40对食管癌组织及配对的癌旁组织(肿瘤边缘距离>3 cm)。采用实时定量逆转录聚合酶链反应(qRT-PCR)检测食管癌组织和癌旁组织中ASB16-AS1和miR-1258的表达。将小干扰RNA阴性对照(si-NC)、ASB16-AS1小干扰RNA(si-ASB16-AS1)、miR阴性对照模拟物(miR-NC)、miR-1258模拟物(miR-1258)、si-ASB16-AS1与抗miR-NC、si-ASB16-AS1与抗miR-1258、si-ASB16-AS1与抗miR-1258分别转染至Eca109细胞。采用甲基噻唑基四氮唑(MTT)法检测细胞活力。采用Transwell实验检测细胞迁移和侵袭能力。通过双荧光素酶报告实验和qRT-PCR验证ASB16-AS1与miR-1258之间的关系。食管癌组织中ASB16-AS1和miR-1258的表达水平分别为2.95±0.27和0.62±0.06。与癌旁组织中的1.00±0.06和1.00±0.07相比,差异具有统计学意义(<0.05)。si-NC组在48 h和72 h的细胞活力分别为0.81±0.07和1.15±0.11,而si-ASB16-AS1组分别为0.46±0.04和0.62±0.06(<0.05)。si-NC组细胞迁移和侵袭数量分别为86.32±8.24和71.29±7.15,而si-ASB16-AS1组分别为43.22±4.31和32.36±3.58,差异具有统计学意义(<0.05)。miR-NC组在48 h和72 h的细胞活力分别为0.84±0.08、1.18±0.12,而miR-1258组分别为0.55±0.05、0.71±0.07(<0.05)。miR-NC组迁移和侵袭数量分别为(83.15±8.31)和(75.33±7.51),而miR-1258组分别为(49.58±4.23)和(38.42±3.84),差异具有统计学意义(<0.05)。si-ASB16-AS1+抗miR-NC组在48 h和72 h的细胞活力分别为0.45±0.04、0.61±0.06,而si-ASB16-AS1+抗miR-1258组分别为0.72±0.07、0.98±0.08;si-ASB16-AS1+抗miR-NC组细胞迁移和侵袭数量分别为44.36±4.41和31.69±3.85,而si-ASB16-AS1+抗miR-1258组分别为72.65±7.27和61.22±6.14,差异具有统计学意义(<0.05)。ASB16-AS1靶向负调控miR-1258的表达。ASB16-AS1在食管癌中上调。ASB16-AS1通过靶向miR-1258促进食管癌细胞的增殖、迁移和侵袭。