环状RNA FBXO11靶向miR-376a-3p/SNRPB轴调控胃癌SNU-1细胞的增殖与凋亡

Circular RNA FBXO11 regulates the proliferation and apoptosis of gastric cancer SNU-1 cells through targeting miR-376a-3p/SNRPB axis

目的:探讨环状RNA FBXO11(circFBXO11)调控miR-376a-3p/小核糖核蛋白多肽B基因(SNRPB)轴对胃癌细胞增殖与凋亡的影响。方法:选取2018年1月至2019年1月华北理工大学附属医院肿瘤外科30例手术切除的胃癌患者的癌和癌旁组织标本,免疫组化染色法检测胃癌组织中SNRPB蛋白阳性表达率,qPCR法检测胃癌组织、胃癌细胞系SNU-1、AGS及HS-746T和胃黏膜细胞GES1中circFBXO11、miR-376a-3p和SNRPB mRNA的表达水平。用双荧光素酶报告基因实验验证circFBXO11与miR-376a-3p、miR-376a-3p与SNRPB之间的靶向关系。将si-NC、si-circFBXO11、miR-NC、miR-376a-3p、si-SNRPB、si-circFBXO11+anti-miR-NC、si-circFBXO11+anti-miR-376a-3p、si-circFBXO11+pcDNA-NC、si-circFBXO11+pcDNA-SNRPB等分别转染进SNU-1细胞,用CCK-8法、流式细胞术以及WB法分别检测细胞的增殖活力、凋亡率以及SNRPB、cyclin D1和C-caspase-3蛋白的表达水平。结果:与癌旁组织比较,胃癌组织中circFBXO11表达水平显著升高、SNRPB蛋白阳性率升高、miR-376a-3p表达显著降低(均P<0.01);与GES1细胞比较,胃癌细胞中circFBXO11和SNRPB表达水平显著升高、miR-376a-3p表达水平显著降低(均P<0.01)。circFBXO11靶向负调控miR-376a-3p表达,miR-376a-3p靶向负调控SNRPB表达。抑制circFBXO11表达或过表达miR-376a-3p或抑制SNRPB表达后,SNU-1细胞的增殖活力降低、凋亡率升高(均P<0.01)。抑制miR-376a-3p表达可部分逆转抑制circFBXO11对SNU-1细胞增殖和凋亡的作用(均P<0.01)。过表达SNRPB可部分逆转抑制circFBXO11对SNU-1细胞增殖和凋亡的影响(均P<0.01)。结论:胃癌组织中circFBXO11呈高表达,抑制circFBXO11表达可抑制胃癌细胞增殖并诱导细胞凋亡,其机制与调控miR-376a-3p/SNRPB通路相关。

Objective:To investigate the effect of circular RNA FBXO11(circFBXO11)regulating the miR-376 a-3 p/SNRPB(small nuclear ribonucleoprotein polypeptides B gene)axis on the proliferation and apoptosis of gastric cancer SNU-1 cells.Methods:Cancer and para-cancerous tissue samples from 30 patients with gastric cancer who underwent surgical resection were surgically resected in the Department of Oncosurgery,the Affiliated Hospital of North China University of Science and Technology from January 2018 to January2019 were collected.The positive expression rate of SNRPB protein in gastric cancer tissues was detected by Immunohistochemical staining.The expression levels of circFBXO11,miR-376 a-3 p and SNRPB mRNA in gastric cancer tissues,gastric cancer cell lines(SNU-1,AGS and HS-746T)and gastric mucosal cell line GES1 were detected by qPCR.Dual luciferase reporter gene assay was used to determine the relationship between circFBXO11 and miR-376a-3p as well as between miR-376 a-3 p and SNRPB.The si-NC,si-circFBXO11,miR-NC,miR-376 a-3p,si-SNRPB,si-circFBXO11+anti-miR-NC,si-circFBXO11+anti-miR-376a-3p,si-circFBXO11+pcDNA-NC,si-circFBXO11+pcDNA-SNRPB were transfected into gastric cancer SNU-1 cells,respectively.CCK-8 assay,Flow cytometry and WB assay were used to detect cell proliferation activity,apoptosis rate and protein expressions of SNRPB,cyclin D1 and C-caspase-3,respectively.Results:Compared with para-cancerous tissues,the expression level of circFBXO11 and the positive rate of SNRPB protein in gastric cancer tissues were significantly increased(all P<0.01),while the expression of miR-376a-3p was significantly decreased(P<0.01).Compared with GES1 cells,the expressions of circFBXO11 and SNRPB were significantly increased,while the expression of miR-376a-3p was significantly decreased(all P<0.01)in gastric cancer cells.circFBXO11 negatively regulated miR-376a-3p expression,and miR-376a-3p negatively regulated SNRPB expression.After inhibiting the expression of circFBXO11 or over-expressing miR-376a-3p or suppressing the expression of SNRPB,the proliferation viability of SNU-1 cells was decreased,and the apoptosis rate was increased(P<0.01).Either inhibiting miR-376 a-3p or over-expressing SNRPB could partially reverse the effect of circFBXO11 suppression on proliferation and apoptosis of SNU-1 cells(all P<0.01).Conclusion:circFBXO11 is highly expressed in gastric cancer tissues.Inhibiting circFBXO11 inhibits the proliferation and induces apoptosis of gastric cancer cells,and the mechanism is related to the regulation of miR-376A-3p/SNRPB pathway.