miR-133靶向JAK2抑制胃癌细胞增殖、迁移和侵袭

MiR-133 inhibits cell proliferation,migration,and invasion in gastric cancer cells by targeting JAK2

目的研究miR-133对胃癌(gastric cancer, GC)细胞增殖、迁移和侵袭的影响,并探讨其作用机制。方法运用qRT-PCR法检测组织和细胞中miR-133、JAK2的mR NA表达;将miR-133组(转染miR-133 mimics)、miR-NC组(未转染细胞)、miR-133 inhibitors组(转染miR-133 inhibitors)、inhibitors-NC组(转染inhibitors)、JAK2WT(载体psiCHECK2-JAK2-32 MUT(载体pUT和miR-133共转染)、miUTRWT和miR-133共转染)、JAKsiCHECK2-JAK2-3UTR MR-133+JAK2组(miR-133 mimics和JAK2共转染)、miR-133+Vector组(miR-133 mimics和pc-DNA 3。1共转染)均用脂质体法转染至AGS、MGC-803细胞;用Western blot检测细胞中JAK2的蛋白表达; MTT法检测细胞增殖;Transwell法检测细胞迁移和侵袭;双荧光素酶报告基因检测实验检测细胞荧光素酶活性。结果与人癌旁组织、人正常胃黏膜细胞GES-1相比, GC组织、GC细胞AGS、MGC-803中miR-133均低表达,JAK2均高表达;且过表达miR-133、沉默JAK2均可抑制GC细胞增殖、迁移和侵袭; JAK2为miR-133的靶点,且回补JAK2可逆转miR-133对GC细胞增殖、迁移和侵袭的抑制作用。结论miR-133可抑制GC细胞增殖、迁移和侵袭,其可能与靶向JAK2有关,将可为GC的临床诊断治疗提供新靶点。

AIM To investigate the role of miR-133 in the proliferation,migration,and invasion of gastric cancer(GC)cells,and to explore the underlying mechanism.METHODS The expression of miR-133 and JAK2 mRNA in tissues and cells was detected by qRT-PCR.AGS and MGC-803 cells were transfected with miR-133 mimic(miR-133 group),miR-133 inhibitor(miR-133 inhibitor group),nonspecific inhibitor(inhibitor-NC group),psiCHECK2-JAK2-3 UTR WT vector and miR-133 mimic(JAK2 WT group),psiCHECK2-JAK2-3 UTR MUT vector and miR-133 mimic(JAK2 MUT group),miR-133 mimic and JAK2(miR-133 + JAK2 group),or miR-133 mimic and pc-DNA 3.1(miR-133 + vector group)using a liposomemediated method.Untransfected cells(miR-NC group)were also included as a control.The protein expression of JAK2 was detected by Western blot.Cell proliferation was detected by MTT assay.Cell migration and invasion were detected by Transwell assay.The luciferase activity was detected by double luciferase reporter assay.RESULTS Compared with human paracancerous tissues or normal gastric mucosal cells(GES-1),miR-133 was down-regulated in GC tissues and GC cells(AGS and MGC-803),and JAK2 was highly expressed in GC tissues and AGS and MGC-803 cells(P<0.05).Overexpression of miR-133 or silencing JAK2 could inhibit cell proliferation,migration,and invasion in GC cells.JAK2 is a target of miR-133,and JAK2 could rescue the inhibitory effect of miR-133 on cell proliferation,migration,and invasion in GC cells.CONCLUSION MiR-133 could inhibit the proliferation,migration,and invasion of GC cells via mechanisms possibly related to targeting of JAK2,which will provide a new target for the clinical diagnosis and treatment of GC.