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

doi:10.16352/j.issn.1001-6325.2023.07.1069

摘要/Abstract

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20 μmol/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。

关键词: 宫颈癌, 山萘酚, 细胞生物行为, VPS9D1-AS1, miR-377-3p 文章编号:1001-6325(2023)07-1069-10

Abstract: 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 (5 μmol/L、10 μmol/L and 20 μmol/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 apoptosis by down regulating VPS9D1-AS1 targeted at miR-377-3p.

Key words: cervical cancer, kaempferol, cell biological behavior, VPS9D1-AS1, miR-377-3p

中图分类号:

R73-36

张禹鑫, 朱紫薇, 张燕, 胡玉红. 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[J]. 基础医学与临床, 2023, 43(7): 1069-1078.

ZHANG Yuxin, ZHU Ziwei, ZHANG Yan, HU Yuhong. Effects of kaempferol regulating VPS9D1-AS1/miR-377-3p on proliferation and apoptosis of cervical cancer cell line SiHa[J]. Basic & Clinical Medicine, 2023, 43(7): 1069-1078.

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