[1]杨良权,于 淼,姜 茜,等.miR-371a-5p靶向抑制LZTS2促进三阴性乳腺癌细胞增殖、迁移、侵袭和上皮间充质转化[J].现代检验医学杂志,2026,41(02):70-76.[doi:10.3969/j.issn.1671-7414.2026.02.012]
　YANG Liangquan,YU Miao,JIANG Qian,et al.MiR-371a-5p Targeted Inhibition of LZTS2 Promoted the Proliferation, Migration, Invasion and Epithelial Mesenchymal Transition of Triple-Negative Breast Cancer Cells[J].Journal of Modern Laboratory Medicine,2026,41(02):70-76.[doi:10.3969/j.issn.1671-7414.2026.02.012]
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miR-371a-5p靶向抑制LZTS2促进三阴性乳腺癌细胞增殖、迁移、侵袭和上皮间充质转化()

《现代检验医学杂志》[ISSN:/CN:]

卷:
第41卷
期数:
2026年02期
页码:
70-76
栏目:
论著
出版日期:
2026-03-15

文章信息/Info

Title:
MiR-371a-5p Targeted Inhibition of LZTS2 Promoted the Proliferation, Migration, Invasion and Epithelial Mesenchymal Transition of Triple-Negative Breast Cancer Cells
文章编号:
1671-7414（2026）02-070-07
作者:
杨良权，于 淼，姜 茜，贺红梅，张 宁
秦皇岛市妇幼保健院乳腺外科，河北秦皇岛 066001
Author(s):
YANG Liangquan，YU Miao，JIANG Qian，HE Hongmei，ZHANG Ning
Department of Breast Surgery, Qinhuangdao Maternal and Child Health Hospital, Hebei Qinhuangdao 066001, China
关键词:
微小核糖核酸-371a-5p; 亮氨酸拉链肿瘤抑制因子2; 三阴性乳腺癌; 增殖; 迁移; 侵袭; 上皮间充质转化
分类号:
R737.9；R730.43
DOI:
10.3969/j.issn.1671-7414.2026.02.012
文献标志码:
A
摘要:
目的?探究微小RNA(miR)-371a-5p靶向调控亮氨酸拉链肿瘤抑制因子2(LZTS2)对三阴性乳腺癌(TNBC)细胞增殖、迁移、侵袭和上皮间充质转化(EMT)的影响。方法收集2022年8月~2024年8月在秦皇岛市妇幼保健院接受根治性手术的60例TNBC患者作为研究对象,收集TNBC组织和邻近的非肿瘤乳腺组织,实时荧光定量PCR(qRT-PCR)检测非肿瘤乳腺组织、TNBC组织和人正常乳腺MCF-10A细胞、TNBC细胞系中LZTS2mRNA的相对表达水平。将MDA-MB-231细胞分为对照组、LZTS2空载质粒组、LZTS2过表达组、miR-371a-5p阴性对照组、miR-371a-5p模拟物组、miR-371a-5p模拟物+LZTS2过表达组。qRT-PCR检测各组LZTS2mRNA和miR-371a-5p相对表达水平;细胞计数试剂盒-8(CCK-8)、划痕愈合实验和Transwell实验分别检测各组MDA-MB-231细胞增殖、迁移和侵袭;蛋白质印迹(Westernblotting)法检测LZTS2和EMT相关蛋白表达;双荧光素酶报告基因实验确认miR-371a-5p与LZTS2之间的靶向关系;裸鼠异种移植肿瘤实验检测小鼠体内miR-371a-5p对LZTS2mRNA表达和肿瘤生长的影响。结果与非肿瘤乳腺组织比较,TNBC组织中LZTS2mRNA(0.33±0.05vs1.00±0.04)表达水平显著降低,差异具有统计学意义(t=81.051,P<0.05);与MCF-10A细胞比较,HCC-1937细胞、MDA-MB-231细胞、MDA-MB-436细胞、MDA-MB-468细胞中LZTS2mRNA(0.44±0.05、0.35±0.04、0.38±0.05、0.53±0.07vs1.00±0.05)表达水平显著降低,差异具有统计学意义(t=15.454~28.712,均P<0.05);且MDA-MB-231细胞中LZTS2mRNA表达水平最低,故选MDA-MB-231细胞进行后续实验。与对照组和LZTS2空载质粒组比较,LZTS2过表达组LZTS2mRNA、LZTS2蛋白、E-cadherin蛋白表达水平显著升高,差异具有统计学意义(q对照组=25.034、13.598、11.714;qLZTS2空载质粒组=23.493、11.993、10.190,均P<0.05),细胞活力、划痕愈合率、侵袭细胞数量、N-cadherin蛋白、Vimentin蛋白、Twist蛋白表达水平显著降低,差异具有统计学意义(q对照组=9.416~13.894,qLZTS2空载质粒组=10.850~15.309,均P<0.05)。miR-371a-5能够特异性靶向LZTS2mRNA3’-非翻译区中的结合位点。与miR-371a-5p阴性对照组比较,miR-371a-5p模拟物组miR-371a-5p、细胞活力、划痕愈合率、侵袭细胞数量、N-cadherin蛋白、Vimentin蛋白、Twist蛋白表达水平显著升高(q=9.034~19.579),LZTS2mRNA、LZTS2蛋白、E-cadherin蛋白表达水平显著降低(q=21.400、15.757、9.617),差异具有统计学意义(均P<0.05);与miR-371a-5p模拟物组比较,miR-371a-5p模拟物+LZTS2过表达组细胞活力、划痕愈合率、侵袭细胞数量、N-cadherin蛋白、Vimentin蛋白、Twist蛋白表达水平显著降低(q=2.648~10.124),LZTS2mRNA、LZTS2蛋白、E-cadherin蛋白表达水平显著升高(q=11.200、17.258、4.909),差异具有统计学意义(均P<0.05)。miR-371a-5p过表达可通过抑制LZTS2mRNA促进裸鼠体内TNBC的肿瘤生长。结论miR-371a-5p可能通过靶向抑制LZTS2表达促进TNBC细胞增殖、迁移、侵袭和EMT。
Abstract:
Objective To explore the effects of miR-371a-5p targeting leucine zipper tumor suppressor 2 (LZTS2) on prolifera-tion, migration, invasion and epithelial-mesenchymal transition (EMT) in triple-negative breast cancer (TNBC) cells. Methods TNBC tissues and adjacent non-tumor breast tissues of 60 patients with TNBC who underwent radical surgery in Qinhuangdao Maternal and Child Health Hospital from August 2022 to August 2024 were collected. Real-time quantitative fluorescent PCR (RT-qPCR) was used to detect the relative expression level of LZTS2 messenger RNA (mRNA) in non-tumor breast tissue, TNBC tissue and human normal breast MCF-10A cells and TNBC cell lines. MDA-MB-231 cells were divided into control group, LZTS2 empty vector plasmid group, LZTS2 overexpression group, miR-371a-5p negative control group, miR-371a-5p mimics group, miR-371a-5p mimics+LZTS2 overexpression group. The relative expression levels of LZTS2 mRNA and miR-371a-5p were detected by RT-qPCR. Cell proliferation, migration and invasion of MDA-MB-231 cells were detected by Cell Counting Kit-8 (CCK-8), scratch healing experiment and Transwell assay, respectively. The expression of LZTS2 and EMT-relat-ed proteins was detected by Western blotting. A dual luciferase reporter assay was used to confirm the targeting relationship be-tween miR-371a-5p and LZTS2. The effects of miR-371a-5p on LZTS2 mRNA expression and tumor growth were detected using nude mouse xenograft tumor models. Results Compared with non-tumor breast tissue, LZTS2 mRNA (0.33±0.05 vs 1.00±0.04) levels in TNBC tissue were significantly decreased (t=81.051, P<0.05). Compared with MCF-10A cells, LZTS2 mRNA (0.44±0.05, 0.35±0.04, 0.38±0.05, 0.53±0.07 vs 1.00±0.05) levels in HCC-1937, MDA-MB-231, MDA-MB-436, and MDA-MB-468 cells were significantly decreased (t=15.454 ~ 28.712, all P<0.05). MDA-MB-231 cells exhibited the lowest LZTS2 mRNA level, thus selected for subsequent experiments. Compared with control group and LZTS2 empty vector group, the levels of LZTS2 mRNA, LZTS2 protein and E-cadherin protein in the LZTS2 overexpression group were significantly in-creased, with statistically significant differences (qcontrol group=25.034, 13.598, 11.714, qLZTS2 empty plasmid group=23.493、11.993、10.190, all P<0.05). Cell viability, scratch healing rate, invasive cell count, and expression levels of N-cadherin, Vimentin and Twist pro-teins were significantly decreased, with statistically significant differences (qcontrol group=9.416~13.894, qLZTS2 empty plasmid group=10.850 ~15.309, all P<0.05). MiR-371a-5 can specifically target the binding site in the 3’-untranslated region of LZTS2 mRNA. Com-pared with miR-371a-5p negative control group, the miR-371a-5p mimic group showed significantly increased levels of miR-371a-5p, cell viability, scratch healing rate, invasive cell number, N-cadherin protein, Vimentin protein and Twist protein expres-sion levels (q=9.034 ~ 19.579, all P<0.05), while the levels of LZTS2 mRNA, LZTS2 protein and E-cadherin protein were significantly decreased (q=21.400, 15.757, 9.617, all P<0.05). Compared with the miR-371a-5p mimics group, cell viability, scratch healing rate, invasive cell count, N-cadherin protein, Vimentin protein and Twist protein expression levels in the miR-371a-5p mimics + LZTS2 overexpression group were significantly decreased (q=2.648 ~ 10.124, all P<0.05), while the levels of LZTS2 mRNA, LZTS2 protein and E-cadherin protein were significantly increased (q=11.200, 17.258, 4.909, all P<0.05). Over-expression of miR-371a-5p can promote TNBC tumor growth in nude mice by inhibiting LZTS2 mRNA. Conclusions MiR-371a-5p may promote TNBC cell proliferation, migration, invasion and EMT through targeted-inhibition of LZTS2 expression.

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备注/Memo

备注/Memo:
基金项目：秦皇岛市科学技术研究与发展计划项目（202301A259）。
作者简介：杨良权（1980-），男，硕士，副主任医师，研究方向：乳腺肿瘤综合治疗，E-mail：yangliangquan6749@163.com。
更新日期/Last Update: 2026-03-15