Department of General Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Department of Neurology, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei, China.
Turk J Gastroenterol. 2023 May;34(5):497-507. doi: 10.5152/tjg.2023.21998.
Colorectal cancer is a common gastrointestinal malignancy worldwide. Many studies have proved that long noncoding RNA alterations participate in colorectal cancer development. This study sought to probe the regulatory mechanism of lncRNA IGF-like family member 2 antisense RNA 1 (IGFL2-AS1) in colorectal cancer cell malignant proliferation.
LncRNA IGFL2-AS1 expression in colorectal cancer cancer and para-cancerous tissues and colorectal cancer cell lines was detected via quantitative real-time polymerase chain reaction. HCT116 cells were transfected with si-IGFL2-AS1, microRNA (miR)- 433-3p inhibitor or p21 (RAC1)-activated kinase 4, PAK4 and IGFL2-AS1 overexpression vector, followed by assessment of cell proliferation and clone formation using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay and colony formation assay. The subcellular localization of lncRNA IGFL2-AS1 was predicted and testified via the nuclear/cytosol fractionation assay. The downstream miRNA of lncRNA IGFL2-AS1 and downstream target of miRNA were predicted, and their binding relationships were testified using dualluciferase assay and RNA immunoprediction experiment.
LncRNA IGFL2-AS1 was abundantly expressed in colorectal cancer tissues and cells. Silencing lncRNA IGFL2-AS1 discouraged HCT116 cell malignant proliferation, while lncRNA IGFL2-AS1 overexpression played an opposite role. LncRNA IGFL2-AS1 was mainly expressed in the cytoplasm of HCT116 cells. LncRNA IGFL2-AS1 bound to miR-433-3p and inhibited its expression. miR-433-3p tar geted PAK4. Silencing lncRNA IGFL2-AS1 facilitated miR-433-3p to suppress PAK4 transcription. miR-433-3p inhibition or PAK4 overexpression partly reversed the inhibition of lncRNA IGFL2-AS1 on HCT116 cell malignant proliferation.
LncRNA IGFL2-AS1 was abundantly expressed in colorectal cancer tissues and cells, and comparatively bound to miR433-3p to facilitate PAK4 transcription, thus promoting HCT116 cell malignant proliferation.
结直肠癌是一种常见的胃肠道恶性肿瘤。许多研究已经证明，长非编码 RNA 的改变参与了结直肠癌的发生发展。本研究旨在探讨 lncRNA IGF 样家族成员 2 反义 RNA 1（IGFL2-AS1）在结直肠癌细胞恶性增殖中的调控机制。
通过实时定量聚合酶链反应检测结直肠癌癌组织和癌旁组织及结直肠癌细胞系中 lncRNA IGFL2-AS1 的表达。用 si-IGFL2-AS1、microRNA（miR）-433-3p 抑制剂或 p21（RAC1）-激活激酶 4（PAK4）、PAK4 和 IGFL2-AS1 过表达载体转染 HCT116 细胞，然后通过 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四唑溴盐测定和集落形成实验评估细胞增殖和克隆形成。通过核/胞质分离实验预测和验证 lncRNA IGFL2-AS1 的亚细胞定位。预测 lncRNA IGFL2-AS1 的下游 miRNA 和 miRNA 的下游靶标，并通过双荧光素酶报告基因实验和 RNA 免疫沉淀实验验证其结合关系。
lncRNA IGFL2-AS1 在结直肠癌组织和细胞中大量表达。沉默 lncRNA IGFL2-AS1 抑制 HCT116 细胞的恶性增殖，而过表达 lncRNA IGFL2-AS1 则起到相反的作用。lncRNA IGFL2-AS1 主要在 HCT116 细胞的细胞质中表达。lncRNA IGFL2-AS1 与 miR-433-3p 结合并抑制其表达。miR-433-3p 靶向 PAK4。沉默 lncRNA IGFL2-AS1 促进 miR-433-3p 抑制 PAK4 转录。miR-433-3p 抑制或 PAK4 过表达部分逆转了 lncRNA IGFL2-AS1 对 HCT116 细胞恶性增殖的抑制作用。
lncRNA IGFL2-AS1 在结直肠癌组织和细胞中大量表达，与 miR433-3p 结合促进 PAK4 转录，从而促进 HCT116 细胞的恶性增殖。
