[摘 要] 目的：探讨lncRNA SOX2-OT是否通过调控miR-215-5p/NIN1/RPN12结合蛋白1同源物（NOB1）信号通路抑制结直 肠癌（CRC）HCT-116细胞增殖和迁移。方法：收集2022年6月至2024年5月期间武汉市金银潭医院收治的29例CRC患者的癌 及癌旁组织标本，以及CRC细胞SW480、HCT-116、HP116和LoVo及人结肠上皮细胞HCoEpiC。qPCR法检测CRC组织和细胞中 SOX2-OT、miR-215-5p和NOB1 mRNA的表达水平。利用RNA干扰技术分别将敲低或过表达SOX2-OT质粒及阴性对照质粒转 染至HCT-116细胞，实验分为对照组、si-NC组、si-SOX2-OT组、si-SOX2-OT + inhibitor（Inh）NC组、si-SOX2-OT + miR-215-5p Inh 组、si-SOX2-OT + oe-NC组、si-SOX2-OT + oe-NOB1组。qPCR法检测各组细胞中SOX2-OT、miR-215-5p和NOB1 mRNA表 达水平，通过MTT、划痕愈合实验、Transwell实验和流式细胞术分别检测各组细胞的增殖、迁移、侵袭及凋亡水平，WB法检 测各组细胞中E-cadherin、N-cadherin、vimentin、Bcl-2、BAX、PCNA、MMP-9和NOB1蛋白表达水平。通过双萤光素酶报告基因 实验验证miR-215-5p与SOX2-OT和NOB1之间的靶向关系。结果：CRC组织和细胞中SOX2-OT和NOB1 mRNA均呈高表达、 miR-215-5p低表达（均P < 0.05）。敲低SOX2-OT表达的HCT-116细胞中SOX2-OT和NOB1 mRNA表达水平、增殖、划痕愈合率 和侵袭细胞数量，以及细胞中N-cadherin、vimentin、Bcl-2、NOB1、PCNA和MMP-9蛋白表达均显著降低（均P < 0.05），miR-215-5p、 细胞凋亡率、E-cadherin和BAX蛋白表达均显著升高（均P < 0.05）。敲低miR-215-5p表达或上调NOB1表达均可减弱敲低 SOX2-OT表达对细胞的抑制作用（均P < 0.05）。miR-215-5p与SOX2-OT和NOB之间存在靶向关系。结论：敲低SOX2-OT表 达可促进miR-215-5p表达，抑制NOB1表达进而抑制HCT-116细胞的增殖、迁移、侵袭并促进细胞凋亡。

[Abstract] Objective: To investigate whether lncRNA SOX2-OT inhibits the proliferation and migration of colorectal cancer (CRC) HCT-116 cells by regulating the miR-215-5p/NIN/RPN12 binding protein 1 homolog (NOB1) signaling pathway. Methods: Cancerous and paired adjacent tissue samples from 29 CRC patients treated at Wuhan Jinyintan Hospital from June 2022 to May 2024 were collected, along with CRC cell lines (SW480, HCT-116, HP116, and LoVo) and normal human colon epithelial HCoApiC cells. The mRNA expression levels of SOX2-OT, miR-215-5p, and NOB1 in CRC tissues and cells were measured using qPCR method. HCT-116 cells were transfected with SOX2-OT knockdown or overexpression plasmids and corresponding negative control plasmids using RNA interference technology, dividing the cells into control group, si-NC group, si-SOX2-OT group, si-SOX2-OT + inhibitor (Inh) NC group, si-SOX2-OT + miR-215-5p Inh group, si-SOX2-OT + oe-NC group, and si-SOX2-OT + oe-NOB1 group. The mRNA expression levels of SOX2-OT, miR-215-5p, and NOB1 in each group of cells were detected using qPCR method. MTT assay, scratch wound healing assay, Transwell chamber assay, and flow cytometry were used to measure cell proliferation, migration, invasion, and apoptosis, respectively. Western blot was applied to detect protein expression levels of E-cadherin, N-cadherin, vimentin, Bcl-2, BAX, PCNA, MMP-9, and NOB1. The targeting relationship between miR-215-5p and SOX2-OT or NOB1 was validated using dual luciferase reporter gene assays. Results: SOX2-OT and NOB1 mRNA were significantly upregulated, while miR-215-5p was downregulated in both CRC tissues and cells (all P < 0.05). In HCT-116 cells with SOX2-OT knockdown, the expression of SOX2-OT and NOB1 mRNA, cell proliferation, wound healing rate, invasive cell number, and protein levels of N-cadherin, vimentin, Bcl-2,NOB1, PCNA, and MMP-9 were significantly reduced (all P < 0.05), while miR-215-5p expression, apoptosis rate, and protein levels of E-cadherin and BAX were significantly increased (all P < 0.05). Both miR-215-5p knockdown and NOB1 overexpression reversed the inhibitory effects of SOX2-OT knockdown on HCT-116 cells (both P < 0.05). miR-215-5p was validated to target SOX2-OT and NOB1. Conclusion: SOX2-OT knockdown upregulates miR-215-5p expression and downregulates NOB1 expression, further inhibiting the proliferation, migration, and invasion of HCT-116 cells and promoting apoptosis.

[基金项目] 武汉市卫生健康委员会资助项目（No. WX17D19）