山萘酚调控VPS9D1-AS1/miR-377-3p对人宫颈癌细胞系SiHa增殖和凋亡的影响  

Effects of kaempferol regulating VPS9D1-AS1/miR-377-3p on proliferation and apoptosis of cervical cancer cell line SiHa

作　　者：张禹鑫 朱紫薇[1] 张燕[1] 胡玉红[1] ZHANG Yuxin;ZHU Ziwei;ZHANG Yan;HU Yuhong(Department of Obstetrics and Gynecology,the First Affiliated Hospital of Jiamusi University,Jiamusi 154000,China)

机构地区：[1]佳木斯大学附属第一医院妇产科,黑龙江佳木斯154000

出　　处：《基础医学与临床》2023年第7期1069-1078,共10页Basic and Clinical Medicine

基　　金：黑龙江省大学生创新创业训练计划项目(201810222067)。

摘　　要：目的探讨山萘酚调控长链非编码RNAVPS9D1反义RNA 1(VPS9D1-AS1)/miR-377-3p对人宫颈癌细胞系增殖和凋亡的影响。方法将人宫颈癌细胞系SiHa分为对照组、山萘酚低、中和高剂量组(5、10和20μmol/L山萘酚)、si-NC组(转染si-NC)、si-VPS9D1-AS1组(转染si-VPS9D1-AS1)、山萘酚+pcDNA-VPS9D1-AS1组(20 mg/L山萘酚+转染pcDNA-VPS9D1-AS1)、山萘酚+anti-miR-377-3p组(20 mg/L山萘酚+转染anti-miR-377-3p)。RT-qPCR、MTT法和集落形成实验分别测定VPS9D1-AS1、miR-377-3p表达、细胞活性和集落形成;流式细胞测量术、划痕试验、Transwell小室法和Western blot分别测定细胞凋亡、迁移、侵袭和Bax及Bcl-2表达;荧光素酶活性检测分析VPS9D1-AS1对miR-377-3p的靶向作用。结果与对照相比,低、中和高剂量山萘酚降低SiHa细胞中VPS9D1-AS1表达水平,集落形成数、细胞活性、Bcl-2蛋白表达水平、迁移距离和侵袭细胞数,且提高了miR-377-3p表达水平、凋亡率和Bax蛋白表达水平;作用均呈浓度依赖性(P<0.05)。SiHa细胞中沉默VPS9D1-AS1后,miR-377-3p表达水平、凋亡率和Bax蛋白表达水平高于si-NC组,且集落形成数、Bcl-2蛋白表达水平、细胞活性、迁移距离和侵袭细胞数低于si-NC组(P<0.05)。VPS9D1-AS1能靶向miR-377-3p。山萘酚处理的SiHa增殖、凋亡、迁移和侵袭的影响能被过表达VPS9D1-AS1或抑制miR-377-3p所逆转。结论山萘酚通过靶向miR-377-3p下调VPS9D1-AS1,减轻宫颈癌细胞系增殖、迁移和侵袭,以及加速凋亡。Objective To investigate the effect of kaempferol on regulation of long non-coding RNA(lncRNA)vps9d1 antisense RNA 1(VPS9D1-AS1)/miR-377-3p and on biological behavior of cervical cancer cells.Methods The cervical cancer cell line SiHa was divided into control group,low,medium and high dose of kaempferol group(5μmol/L、10μmol/L and 20μmol/L kaempferol),si-NC group(transfected si-NC),si-VPS9D1-AS1 group(transfected si-VPS9D1-AS1),kaempferol+pcDNA-VPS9D1-AS1 group(20 mg/L kaempferol+transfected pcDNA-VPS9D1-AS1),kaempferol+anti-miR-377-3p group(20 mg/L kaempferol+transfected anti-miR-377-3p).VPS9D1-AS1,miR-377-3p expression,cell activity and colony formation were measured by RT-qPCR,MTT assay and colony formation assay;Cell apoptosis,migration,invasion and Bax/Bcl-2 expression were measured by flow cytometry,scratch test,Transwell test and Western blot.Luciferase activity assay was used to evaluate the targeting effect of VPS9D1-AS1 on miR-377-3p.Results Compared with the control,low,medium and high doses of kaempferol decreased the expression level of VPS9D1-AS1,the number of colony formation,cell activity,Bcl-2 protein expression level,migration distance and the number of invasive cells in SiHa cells,and increased the expression level of miR-377-3p,apoptosis rate as well as Bax protein expression level in a concentration dependent manner(P<0.05).After VPS9D1-AS1 was silenced in SiHa cells,the expression level of miR-377-3p,apoptosis rate and Bax protein expression level were higher than those in si-NC group;The colony forming number,Bcl-2 protein expression,cell activity,migration distance and the accounting of invasive cells in si-NC group were significantly(P<0.05)lower than those in si-NC group.VPS9D1-AS1 targeted at miR-377-3p.The effects of kaempferol treated SiHa on proliferation,apoptosis,migration and invasion were reversed by over-expression of VPS9D1-AS1 or inhibition of miR-377-3p.Conclusions Kaempferol can inhibit the proliferation,migration and invasion of cervical cancer cells and accelerate apop

关 键 词：宫颈癌 山萘酚 细胞生物行为 VPS9D1-AS1 miR-377-3p 

分 类 号：R73-36[医药卫生—肿瘤]