miR-372-3p靶向ADAM9对恶性周围神经鞘瘤细胞增殖、迁移及侵袭的影响

The effects of miR-372-3p on the proliferation, migration and invasion of malignant peripheral nerve sheath tumor cells by targeting ADAM9

摘要:

目的：探讨微小RNA-372-3p（miR-372-3p）靶向解整合素-金属蛋白酶9（ADAM9）对恶性周围神经鞘瘤（MPNST）细胞增殖、迁移及侵袭的影响。方法：qRT-PCR法检测我院2023年6月~2025年6月期间收治的MPNST患者术中切除的MPNST组织中miR-372-3p、ADAM9 mRNA的表达。RIP和双荧光素酶报告基因实验检测miR-372-3p与ADAM9的互作。将ST88-14细胞随机分为NC组、mimic-NC组、miR-372-3p-mimic组、miR-372-3p-mimic+OE-NC组、miR-372-3p-mimic+OE-ADAM9组，采用qRT-PCR评估转染效率。EdU染色和CCK8法检测ST88-14细胞增殖；划痕实验检测ST88-14细胞迁移；Transwell实验ST88-14细胞侵袭；Western blot检测ST88-14细胞中Ki-67、ADAM9、CD44蛋白表达。移植瘤实验验证miR-372-3p过表达对MPNST移植瘤生长和miR-372-3p、ADAM9 mRNA表达的影响。结果：与癌旁组织相比，MPNST组织中miR-372-3p低表达，ADAM9 mRNA高表达（P<0.05）。miR-372-3p可以靶向负调控ADAM9。miR-372-3p-mimic组EdU阳性率、A450值、划痕愈合率、侵袭数、Ki-67、ADAM9 mRNA和蛋白、CD44低于NC组、mimic-NC组，miR-372-3p高于NC组、mimic-NC组（P<0.05）；miR-372-3p-mimic+OE-ADAM9组EdU阳性率、A450值、划痕愈合率、侵袭数、Ki-67、ADAM9 mRNA和蛋白、CD44高于miR-372-3p-mimic组、miR-372-3p-mimic+OE-NC组（P<0.05）。miR-372-3p过表达组MPNST移植瘤重量、体积、ADAM9表达低于对照组，miR-372-3p表达高于对照组（P<0.05）。结论：miR-372-3p通过靶向ADAM9抑制MPNST细胞增殖、迁移和侵袭。

Abstract:

Objective: To discuss the effects of microRNA-372-3p (miR-372-3p) targeting a disintegrin and metalloproteinase 9 (ADAM9) on the proliferation, migration and invasion of malignant peripheral nerve sheath tumor (MPNST) cells. Methods: The expression of miR-372-3p and ADAM9 mRNA in the MPNST tissues resected during the operation of MPNST patients admitted to our hospital from June 2023 to June 2025 were detected by qRT-PCR. The interaction between miR-372-3p and ADAM9 was detected by RIP and dual-luciferase reporter gene assays. ST88-14 cells were randomly assigned into the NC group, mimic-NC group, miR-372-3p-mimic group, miR-372-3p-mimic+OE-NC group, and miR-372-3p-mimic+OE-ADAM9 group. The transfection efficiency was evaluated by qRT-PCR. The proliferation of ST88-14 cells was detected by EdU staining and CCK8 assay. The scratch assay was used to detect the migration of ST88-14 cells. Transwell assay was used to detect ST88-14 cell invasion. Western blot was used to detect the protein expression of Ki-67, ADAM9 and CD44 in ST88-14 cells. The transplanted tumor experiment was used to verify the effect of miR-372-3p overexpression on the growth of MPNST transplanted tumors and the expression of miR-372-3p and ADAM9 mRNA. Results: Compared with adjacent tissues, miR-372-3p was lowly expressed and ADAM9 mRNA was highly expressed in MPNST tissues (P<0.05). MiR-372-3p can target and negatively regulate ADAM9. The EdU positive rate, A450 value, scratch healing rate, invasion number, Ki-67, ADAM9 mRNA and protein, and CD44 in the miR-372-3p-mimic group were lower than those in the NC group and the mimic-NC group, while miR-372-3p was higher than that in the NC group and the mimic-NC group (P<0.05). The EdU positive rate, A450 value, scratch healing rate, invasion number, Ki-67, ADAM9 mRNA and protein, and CD44 in the miR-372-3p-mimic+OE-ADAM9 group were higher than those in the miR-372-3p-mimic group and the miR-372-3p-mimic+OE-NC group (P<0.05). The weight, volume of MPNST transplanted tumors, and the expression of ADAM9 in the miR-372-3p overexpression group were lower than those in the control group, while the expression of miR-372-3p was higher than that in the control group (P<0.05). Conclusion: MiR-372-3p inhibits the proliferation, migration and invasion of MPNST cells by targeting ADAM9.