lncRNA PSMA3-AS1 靶向 miR-3619-5p 调控宫颈癌 SiHa 细胞增殖、迁移及侵袭的分子机制研究

doi:10.3870/j.issn.1672-8009.2023.01.005

摘要/Abstract

摘要： 目的探讨 lncRNA PSMA3-AS1 调控宫颈癌细胞增殖、迁移及侵袭的分子机制。方法收集 2020 年 3 月至 2021 年 6 月西安医学院第二附属医院收治的 42 例宫颈癌患者的癌组织及其癌旁组织标本, 采用 qRT-PCR 法检测宫颈癌组织、癌旁组织、人宫颈上皮永生化细胞 H8、与人宫颈癌细胞系 SiHa、 HeLa、 Caski 中 lncRNA PSMA3-AS1、 miR-3619-5p 的表达量; 以 SiHa 细胞为研究对象, 分组为: si-NC 组、 si-lncRNA PSMA3-AS1 组、 miR-NC 组、 miR-3619-5p 组、 anti-miR-NC + si-lncRNA PSMA3-AS1 组、 anti-miR-3619-5p + si-lncRNA PSMA3-AS1 组; MTT 法与 Transwells 实验分别检测每组 SiHa 细胞的增殖活力、迁移及侵袭能力; 双荧光素酶报告实验检测 miR-3619-5p 过表达对野生型载体 lncRNA PSMA3-AS1-WT、突变型载体 lncRNA PSMA3-AS1-MUT 荧光素酶活性的影响; Western 印迹检测 MMP2、 MMP9 蛋白表达量。结果宫颈癌组织中 lncRNA PSMA3-AS1 的表达量比癌旁组织增加 ( P < 0. 01), miR-3619-5p 的表达量比癌旁组织减少 ( P < 0. 01); 与 H8 细胞比较, SiHa、 HeLa、 Caski 细胞中 lncRNA PSMA3-AS1 的表达量升高 (P< 0. 01), miR-3619-5p 的表达量降低 (P< 0. 01); 与 si-NC 组比较, si-lncRNA PSMA3-AS1 组细胞活力和 MMP2、 MMP9 蛋白水平降低 (P< 0. 05), 迁移及侵袭细胞数减少 (P< 0. 05); 与 miR-NC 组比较, miR-3619-5p 组细胞活力和 MMP2、 MMP9 蛋白水平降低 (P< 0. 01), 迁移及侵袭细胞数减少 (P< 0. 05); miR-3619-5p 过表达可抑制 lncRNA PSMA3-AS1-WT 的荧光素酶活性 (P< 0. 01), 而未能影响 lncRNA PSMA3-AS1-MUT 的荧光素酶活性; 与 anti-miR-NC + si-lncRNA PSMA3-AS1 组比较, anti-miR-3619-5p + si-lncRNA PSMA3-AS1 组细胞活力和 MMP2、 MMP9 蛋白水平升高 (P< 0. 01), 迁移及侵袭细胞数增多 (P< 0. 01)。结论干扰 lncRNA PSMA3- AS1 表达可通过促进 miR-3619-5p 而降低宫颈癌细胞增殖、迁移及侵袭能力。

关键词: 宫颈癌, 增殖, 迁移, 侵袭, lncRNA PSMA3-AS1, miR-3619-5p

Abstract: Objective To explore the molecular mechanism of lncRNA PSMA3-AS1 on regulating the proliferation, migration and invasion of cervical cancer cells. Methods Collect cancer tissue and paracancerous tissue specimens of 42 patients with cervical cancer admitted to the Second Affiliated Hospital of Xi’an Medical University from March 2020 to June 2021. The qRT-PCR method was used to detect the expression levels of lncRNA PSMA3-AS1, miR-3619-5p in cervical cancer tissues, adjacent tissues, human cervical epithelial immortalized cells H8, and human cervical cancer cell lines SiHa, HeLa, and Cashi. Taking SiHa cells as the research object, they were randomly grouped into 6 groups: si-NC group, si-lncRNA PSMA3-AS1 group, miR-NC group, miR-3619-5p group, anti-miR-NC + si-lncRNA PSMA3-AS1 group, anti-miR-3619-5p + si-lncRNA PSMA3-AS1 group. The proliferation, migration and invasion abilities of SiHa cells in each group were assayed by MTT assay and transwell assay respectively. The dual luciferase reporter assay was used to determine the effect of miR-3619-5p overexpression on the luciferase activity of wild-type vector lncRNA PSMA3-AS1-WT and mutant vector lncRNA PSMA3-AS1-MUT. Western blotting was used to detect the protein expression levels of MMP2 and MMP9. Results The expression level of lncRNA PSMA3-AS1 in cervical cancer tissues was higher than that in the adjacent tissues (P< 0. 01), and the expression level of miR-3619-5p was lower than that in the adjacent tissues (P< 0. 01). Compared with the H8 cells, the expression level of lncRNA PSMA3-AS1 in SiHa, HeLa, and Caski cells was increased (P < 0. 01), while the expression level of miR-3619-5p was decreased (P < 0. 01). Compared with the si-NC group, the cell viability, and the protein expression levels of MMP2, MMP9 in the si-lncRNA PSMA3-AS1 group were decreased (P< 0. 05), and the numbers of migrated and invaded cells were decreased (P < 0. 05). Compared with the miR-NC group, the cell viability and the protein expression levels of MMP2, MMP9 in the miR-3619-5p group were decreased (P< 0. 01), and the numbers of migrated and invaded cells were decreased (P < 0. 01). Overexpression of miR-3619-5p could inhibit the luciferase activity of lncRNA PSMA3-AS1-WT (P< 0. 01), but failed to affect the luciferase activity of lncRNA PSMA3-AS1-MUT. Compared with the anti-miR-NC + si-lncRNA PSMA3-AS1 group, the cell viability, and the protein expression levels of MMP2, MMP9 in the anti-miR-3619-5p + si-lncRNA PSMA3-AS1 group were increased (P< 0. 01), and the numbers of migrated and invaded cells were increased (P< 0. 01). Conclusion Interfering with the expression of lncRNA PSMA3-AS1 could reduce the proliferation, migration and invasion of cervical cancer cells by targeting miR-3619-5p.

Key words: cervical cancer, proliferation, migration, invasion, lncRNA PSMA3-AS1, miR-3619-5p

中图分类号:

R737. 33

鲁慧玲, 闫飞艳 , 常丽花. lncRNA PSMA3-AS1 靶向 miR-3619-5p 调控宫颈癌 SiHa 细胞增殖、迁移及侵袭的分子机制研究[J]. 医学分子生物学杂志, 2023, 20(1): 26-33.

LU Huiling , YAN Feiyan , CHANG Lihua . lncRNA PSMA3-AS1 Regulate the Proliferation, Migration and Invasion of Cervical Cancer Cells by Targeting miR-3619-5p[J]. Journal of Medical Molecular Biology, 2023, 20(1): 26-33.

[1]	黄琦, 翟海程. lncRNA CERS6-AS1 通过调控 miR-138-2-3p 抑制人胶质瘤细胞系增殖 [J]. 医学分子生物学杂志, 2023, 20(1): 20-25.
[2]	熊嘉璐∗ , 杨浩∗ , 周斌∗ , 吴朝妍 , 黄金玲 , 周莹. 基于 TCGA 数据库筛选、分析并验证宫颈癌生物标志物 [J]. 医学分子生物学杂志, 2023, 20(1): 49-55.
[3]	郁昊 , 万炳年 , 刘艳萍. CHRDL1 调节结直肠癌增殖、迁移和侵袭[J]. 医学分子生物学杂志, 2023, 20(1): 63-69.
[4]	宋平辉 , 张毅 , 吴忠坤 , 李春博 , 李勤新. HBV HBx 蛋白调控肝癌细胞增殖的机制研究[J]. 医学分子生物学杂志, 2023, 20(1): 70-77.
[5]	叶玉锦, 黄丽珊, 黄巧如, 吴明秀. lncRNA SNHG17 靶向 miR-525-5p 对人绒毛膜滋养层细胞增殖、迁移和侵袭的影响 [J]. 医学分子生物学杂志, 2022, 19(6): 452-456.
[6]	刘昕, 李天客 , 杜宁 , 路月亭 , 刘学聪. CD168 对口腔鳞状细胞癌增殖、侵袭的作用机制研究 [J]. 医学分子生物学杂志, 2022, 19(6): 457-463.
[7]	蒋君霞 , 杨国荣 , 谷涉群 , 喻珣. circ_ 0061140 靶向 miR-6838-5p 促进非小细胞肺癌细胞的增殖、阻滞细胞周期、诱导细胞凋亡[J]. 医学分子生物学杂志, 2022, 19(6): 464-471.
[8]	杨三虎, 李岩. 沉默 NEK2 通过调节 Wnt 信号通路抑制非小细胞肺癌的细胞增殖[J]. 医学分子生物学杂志, 2022, 19(6): 472-477.
[9]	董凡 , 刘海英 , 曾艳. PPAR-γ 过表达缓解肾缺血再灌注大鼠纤维化和炎性因子的异常表达 [J]. 医学分子生物学杂志, 2022, 19(6): 478-483.
[10]	李鲲, 谢陆莉, 邓琳. 黄芪多糖对人牙周膜干细胞生物学特性的影响[J]. 医学分子生物学杂志, 2022, 19(6): 496-500.
[11]	冯堃 , 夏似龙 , 邢兆辉. miR-133b 靶向 PTBP1 对肾细胞癌增殖和侵袭的影响[J]. 医学分子生物学杂志, 2022, 19(5): 353-359.
[12]	郭秀莲 , 邢海 , 王巍. miR-766-5p 过表达介导 MMP-7 对胶质瘤细胞增殖、侵袭与凋亡的影响及机制 [J]. 医学分子生物学杂志, 2022, 19(5): 360-365.
[13]	黄耘 , 于永洋 , 苏蕊 , 王思力. lncRNA HOXB-AS3 通过激活 PI3K-AKT 通路加剧急性髓样白血病患者细胞的增殖、凋亡、迁移和侵袭 [J]. 医学分子生物学杂志, 2022, 19(5): 366-373.
[14]	张强, 霍占江, 田亮, 王志杰, 郝敏丽. 细胞角蛋白 19 通过调控 MAPK 和 PTEN 通路促进甲状腺癌细胞的增殖、侵袭及转移[J]. 医学分子生物学杂志, 2022, 19(5): 399-402.
[15]	黄委珍, 张敬雷, 苏晓平, 孔丽青, 陈惠. 薯蓣皂苷元在口腔鳞癌中的抗癌作用研究 [J]. 医学分子生物学杂志, 2022, 19(5): 403-408.

lncRNA PSMA3-AS1 靶向 miR-3619-5p 调控宫颈癌 SiHa 细胞增殖、迁移及侵袭的分子机制研究

lncRNA PSMA3-AS1 Regulate the Proliferation, Migration and Invasion of Cervical Cancer Cells by Targeting miR-3619-5p

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价