柚皮苷通过上调miR-628-5p抑制宫颈癌ME-180细胞增殖和周期进展  

Study on the mechanism of naringin affecting the proliferation and cycle of cervical cancer ME-180 cells by up regulating miR-628-5p

作　　者：杨瑞英[1] 谢孟珍[1] 梁秀兰 YANG Rui-ying;XIE Meng-zhen;LIANG Xiu-lan(Department of Gynecology People’s Hospital of Xingtai,Xingtai 054001,Hebei province,China)

机构地区：[1]邢台市人民医院妇科,河北邢台054001

出　　处：《中国临床药理学杂志》2022年第4期318-322,共5页The Chinese Journal of Clinical Pharmacology

摘　　要：目的探讨柚皮苷影响宫颈癌ME-180细胞增殖和周期的机制。方法将宫颈癌ME-180细胞分成宫颈癌ME-180细胞分成对照组(正常培养)、实验组(64μmol·L^(-1)的柚皮苷处理)、实验组+Anti-miR-NC组(64μmol·L^(-1)的柚皮苷+转染inhibitor control处理)、实验组+Anti-miR-628-5p组(64μmol·L^(-1)的柚皮苷+转染miR-628-5p inhibitor处理)。转染前将对数期的ME-180细胞浓度调整为每毫升5×10;个,接种于无菌细胞培养板,细胞融合度至75%~85%时,用0.5%胰蛋白酶消化,用Lipofectamine;2000转染试剂将inhibitor control、miR-628-5p inhibitor转染进细胞。用实时定量反转录聚合酶链反应方法检测miR-628-5p表达;用噻唑蓝法测定各组细胞增殖活力;用碘化丙啶单染法检测周期;用Annexin V-FITC/PI双染法检测细胞凋亡;用蛋白质印迹法检测蛋白表达情况。结果对照组、实验组、实验组+Anti-miR-NC组、实验组+Anti-miR-628-5p组宫颈癌ME-180细胞中miR-628-5p表达量为1.00±0.11,1.99±0.16,2.05±0.23和1.05±0.12;细胞增殖活力分别为0.81±0.12,0.42±0.05,0.40±0.06和0.59±0.06;G0/G1期细胞比例为(48.97±5.42)%,(71.93±4.73)%,(71.18±5.77)%和(59.18±5.89)%;细胞凋亡率为(4.19±0.35)%,(18.88±2.01)%,(17.75±1.55)%和(9.76±0.92)%。上述指标,实验组与对照组比较,差异均有统计学意义(均P<0.05);实验组+Anti-miR-628-5p组与实验组+Anti-miR-NC组比较,差异均有统计学意义(均P<0.05)。结论柚皮苷通过上调miR-628-5p抑制宫颈癌ME-180细胞的增殖和周期进展,促进细胞凋亡。Objective To investigate the effect of naringin on the proliferation and cycle of cervical cancer ME-180 cells.Methods Cervical cancer ME-180 cells were divided into control group (cultured normally),experimental group (treated with 64μmol·L^(-1)naringin),experimental+anti-miR-NC group (transfected with inhibitor control and treated with 64μmol·L^(-1)naringin),experimental+anti-miR-628-5p group (transfected with mir-628-5p inhibitor and treated with 64μmol·L^(-1)naringin).Before transfection,ME-180 cells at logarithmic stage were inoculated into sterile cell culture plates at an adjusted concentration of 5×10;.When the degree of cell fusion reached75%~85%,the cells were digested with 0.5% trypsin.The cells were transfected with inhibitor control and miR-628-5p inhibitor using Lipofectamine;2000 transfection reagent.The miR-628-5p expression was detected by qRT-PCR,cell proliferation activity was detected by MTT,cell cycle was detected by PI single staining,cell apoptosis was detected by Annexin V-FITC/PI double staining,protein expression was detected by Western blot.Results The expression of miR-628-5p in control group,experimental group,experimental+anti-miR-NC group,experimental+anti-miR-628-5p group were 1.00±0.11,1.99±0.16 and2.05±0.23,1.05±0.12;the cell proliferation activity were 0.81±0.12,0.42±0.05,0.40±0.06 and0.59±0.06;the proportion of cells in G0/G1 phase were (48.97±5.42)%,(71.93±4.73)%,(71.18±5.77)%and (59.18±5.89)%;the cell apoptosis rate were (4.19±0.35)%,(18.88±2.01)%,(17.75±1.55)%and (9.76±0.92)%.Comparison the above indicators between experimental group and control group,the difference was significant (P<0.05);compared between experimental+anti-miR-628-5p group and experimental+anti-miR-NC group,the difference was significant (P<0.05).Conclusion Naringin inhibits the proliferation,cycle progression and promotes cell apoptosis of cervical cancer ME-180 cells by up-regulating miR-628-5p.

关 键 词：宫颈癌 柚皮苷 凋亡 增殖 miR-628-5p 

分 类 号：R97[医药卫生—药品]