Circ_ZFYVE1对胰岛β细胞增殖、迁移和凋亡的影响  

Effect of Circ_ ZFYVE1 on the proliferation, migration and apoptosis of pancreatic islet β cells

作　　者：孙媛 常向云[2] 朱余蓉[2] 巴涛 SUN Yuan;CHANG Xiangyun;ZHU Yurong(School of Medicine,Shihezi University,Shihezi 832000,China)

机构地区：[1]石河子大学医学院,832000 [2]石河子大学第一附属医院内分泌代谢科

出　　处：《中国糖尿病杂志》2022年第3期221-227,共7页Chinese Journal of Diabetes

基　　金：国家自然科学基金(81860149)。

摘　　要：目的观察Circ;FYVE1(cZFYVE1)对小鼠胰岛β细胞瘤(MIN6)增殖、迁移和凋亡的影响。方法 MIN6细胞分为正常对照组(NC)和高糖组(HG)。NC、HG组分别在含有5.5、25 mmol/L葡萄糖的DMEM完全培养基中培养48 h。q RT-PCR检验cZFYVE1表达。Sanger测序对cZFYVE1反向剪接位点的PCR产物进行验证。慢病毒转染实验分为空载体组(Con)组、敲低环状质粒载体组(cZFYVE1-si)和过表达环状质粒载体组(cZFYVE1-OE)。Con组使用空载体进行转染,cZFYVE1过表达和敲低环状质粒载体转染MIN6胰岛β细胞株。qRT-PCR检测转染后各组MIN6细胞系中c ZFYVE1表达水平;CCK8法、EDU法检测细胞增殖;划痕实验检测各组24 h划痕愈合率;流式细胞术检测细胞凋亡率,Westernblot法检测Caspase-3蛋白表达水平。结果 HG组cZFYVE1表达低于NC组[(0.7169±0.1525)vs(1.0000±0.0000),P<0.05]。Sanger测序证实cZFYVE1反向剪接位点。与Con组比较,cZFYVE1-si组cZFYVE1表达、EDU阳性率、24 h及48 h细胞增殖活力、24 h划痕愈合率降低(P<0.05),cZFYVE1-OE组cZFYVE1表达、EDU阳性率、24 h及48 h增殖活力、24 h划痕愈合率升高(P<0.05)。与Con组比较,cZFYVE1-si组MIN6细胞凋亡率、Caspase-3蛋白表达升高(P<0.05),cZFYVE1-OE组MIN6细胞凋亡率、Caspase-3蛋白表达降低(P<0.05)。结论 cZFYVE1可促进MIN6胰岛β细胞增殖、迁移并减少凋亡。Objective To investigate the effect of Circ_ZFYVE1(cZFYVE1)on the proliferation,migration and apoptosis of mouse pancreatic β-cell tumor(MIN6).Methods MIN6 cells were divided into normal control group(NC)and high glucose group(HG). The NC and HG groups were cultured in DMEM complete medium containing 5. 5 and 25 mmol/L glucose for 48 h,respectively. The expression of cZFYVE1 was tested by q RT-PCR. Sanger sequencing was used to verify the PCR product of the cZFYVE1 reverse splicing site. Lentiviral transfection experiments were divided into empty vector group(Con)group,knockdown circular plasmid vector group(cZFYVE1-si)and overexpression circular plasmid vector group(cZFYVE1-OE). The Con group used empty vector for transfection,and cZFYVE1 overexpression and knockdown circular plasmid vector was transfected into MIN6 pancreatic islet β cell line. The expression level of cZFYVE1 in the MIN6 cell lines of each group was evaluated by q RT-PCR after transfection.CCK8 and EDU methods were used to detect cell proliferation. The scratch test was used to detect the scratch healing rate in each group at 24 h. The cell apoptosis rate was detected by flow cytometry. Western blot method was used to detect the expression level of Caspase-3 protein.Results The expression of cZFYVE1 was lower in HG group than in NC group[(0. 7169 ± 0. 1525)vs(1. 0000 ± 0. 0000),P<0. 05]. Sanger sequencing confirmed the reverse splice site of cZFYVE1. Compared with Con group,the cZFYVE1 expression,EDU positive rate,24,48 h cell proliferation,24 h scratch healing rate decreasedin cZFYVE1-si group(P<0. 05),while cZFYVE1 expression,EDU positive rate,proliferation activity at24,48 h and scratch healing rate at 24 h increased in cZFYVE1-OE group(P<0. 05). Compared with Con group,the apoptosis rate and Caspase-3 protein expression of MIN6 cells increased in cZFYVE1-si group(P<0. 05),while the apoptosis rate and Caspase-3 protein expression of MIN6 cells decreased in cZFYVE1-OE group(P<0. 05).Conclusion cZFYVE1 can promote the proliferation and mig

关 键 词：Circ_ZFYVE1 小鼠胰岛β细胞瘤 增殖 迁移 凋亡 

分 类 号：R587.1[医药卫生—内分泌]