circNHSL1通过调控miR-125b-5p/HMGB3轴抑制人乳腺癌细胞系增殖及促凋亡

doi:10.16352/j.issn.1001-6325.2024.12.1678

摘要/Abstract

摘要： 目的探讨circNHSL1调控miR-125b-5p/HMGB3轴对乳腺癌细胞生物行为的影响。方法将乳腺癌细胞系T47D分为si-NC组、si-circNHSL1组、miR-NC组、miR-125b-5p mimic组、si-circNHSL1+miR-125b-5p inhibitor组。RT-qPCR测定circNHSL1和miR-125b-5p在乳腺癌组织和癌旁组织中的表达。Western blot检测高迁移率族蛋白3(HMGB3)蛋白的表达。双荧光素酶报告基因实验确定circNHSL1与miR-125b-5p、miR-125b-5p与HMGB3之间的相互作用。CCK-8法检测增殖抑制率;集落形成实验检测集落形成数;流式细胞术检测凋亡率;Transwell实验检测迁移和侵袭细胞数。结果与癌旁组织比较,乳腺癌组织中circNHSL1和HMGB3表达升高(P＜0.05),miR-125b-5p相对水平显著降低(P＜0.05)。miR-125b-5p分别与circNHSL1、HMGB3直接结合。circNHSL1敲低下调T47D细胞中circNHSL1和HMGB3表达,上调miR-125b-5p表达(P＜0.05);而miR-125b-5p mimic转染后上调miR-125b-5p表达,下调HMGB3表达(P＜0.05);且miR-125b-5p inhibitor转染能够逆转circNHSL1沉默对miR-125b-5p以及HMGB3表达的影响。circNHSL1低表达或miR-125b-5p高表达增加T47D细胞抑制率、集落形成数以及细胞凋亡率,减少迁移数、侵袭数(P＜0.05);且miR-125b-5p inhibitor的转染挽救circNHSL1沉默对细胞表型的影响(P＜0.05)。结论干扰circNHSL1通过上调miR-125b-5p/HMGB3轴可促进乳腺癌细胞凋亡,抑制其增殖、迁移和侵袭。

关键词: 乳腺癌, circNHSL1, miR-125b-5p, 高迁移率族蛋白3

Abstract: Objective To investigate the regulation of miR-125b-5p/HMGB3 axis by circNHSL1 on the biological behavior of breast cancer cells. Methods The breast cancer cells T47D were divided into si-NC group, si-circNHSL1 group, miR-NC group, and miR-125b-5p mimic group and si-circNHSL1+miR-125b-5p inhibitor group. The expression of circNHSL1 and miR-125b-5p in breast cancer tissues and adjacent tissues was determined using RT-qPCR. Western blot was conducted to detect the protein expression of high mobility group protein 3 (HMGB3). Dual luciferase reporter gene assay was used to confirm the interaction between circNHSL1 and miR-125b-5p, miR-125b-5p and HMGB3. The proliferation rate was measured by CCK-8 method; the colony formation assay was applied to detect the colony formation numbers; flow cytometry was used to measure cell apoptosis rate; Transwell assay was used to detect the numbers of migratory and invasive cells. Results Compared with adjacent tissues, circNHSL1 and HMGB3 expression in breast cancer tissues was significantly increased (P＜0.05), while the expression of miR-125b-5p was significantly decreased (P＜0.05). miR-125b-5p directly bound to circNHSL1 and HMGB3, respectively. The circNHSL1 knockdown down regulated circNHSL1 and HMGB3 expression and upregulated miR-125b-5p expression in T47D cells (P＜0.05). After transfection with miR-125b-5p mimic, miR-125b-5p expression was up-regulated and HMGB3 expression was down-regulated (P＜0.05). Moreover, transfection of miR-125b-5p inhibitor reversed the effect of circNHSL1 silencing on the expression of miR-125b-5p and HMGB3. Low expression of circNHSL1 or high expression of miR-125b-5p increased the cell inhibition rate, colony formation and apoptosis rate of T47D and decreased the number of migratory and invasive cells (P＜0.05). Moreover, transfection of miR-125b-5p inhibitor saved the effect of circNHSL1 silencing on cell phenotypes (P＜0.05). Conclusions Interfering with circNHSL1 might promote cell apoptosis and inhibit cell proliferation, migration and invasion in breast cancer cell by up-regulating the miR-125b-5p/HMGB3 axis.

Key words: breast cancer, circNHSL1, miR-125b-5p, high mobility group protein 3(HMGB3)

中图分类号:

R730.2

李欢, 谢贤鑫. circNHSL1通过调控miR-125b-5p/HMGB3轴抑制人乳腺癌细胞系增殖及促凋亡[J]. 基础医学与临床, 2024, 44(12): 1678-1684.

LI Huan, XIE Xianxin. circNHSL1 inhibits proliferation and promotes apoptosis of human breast cancer cell line through regulating miR-125b-5p/HMGB3 axis[J]. Basic & Clinical Medicine, 2024, 44(12): 1678-1684.

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