MiR-204通过靶向调控HNRNPA2B1抑制乳腺癌的侵袭和转移 |
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引用本文: | 张丽萍,白 俊,胡雅琼,周丹丹,郑 荃,尹崇高,牟青杰,李洪利.MiR-204通过靶向调控HNRNPA2B1抑制乳腺癌的侵袭和转移[J].南方医科大学学报,2020,40(6):869-875. |
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作者姓名: | 张丽萍 白 俊 胡雅琼 周丹丹 郑 荃 尹崇高 牟青杰 李洪利 |
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摘 要: | 目的 探讨miR-204通过靶向调控HNRNPA2B1对乳腺癌侵袭和转移的影响。方法 生物信息学数据库查询miR-204在乳腺癌患者中的表达情况和 miR-204对乳腺癌患者生存率的影响;RT-qRCR检测miR-204在乳腺癌细胞系中的表达情况;利用 GV369-miR-204过表达慢病毒上调MDA-MB-231细胞中的miR-204;Transwell实验检测miR-204对MDA-MB-231细胞侵袭和迁移能力的影响;生物信息学方法确定miR-204的关键基因(Hub基因);双荧光素酶实验检测miR-204和HNRNPA2B1的靶向关系;Western blot实验检测过表达miR-204后HNRNPA2B1的表达情况;Transwell实验检测MDA-MB-231/miR-204+NC、MDA-MB-231/miR-204+HNRNPA2B1细胞的侵袭和迁移能力的变化。结果 miR-204在乳腺癌组织中表达降低(P<0.05),低表达水平的miR-204与患者不良预后相关(P<0.05);miR-204在乳腺癌细胞系MDA-MB-231细胞系中表达量低于正常乳腺上皮细胞MCF-10A的表达(P<0.0001)。通过过表达慢病毒上调MDA-MB-231细胞中miR-204的表达,结果显示上调miR-204可抑制乳腺癌细胞的侵袭和迁移能力(P<0.05);HNRNPA2B1为miR-204的Hub基因,starbase网站发现miR-204-5p
和HNRNPA2B1表达呈负相关且HNRNPA2B1在乳腺癌患者中表达升高(P<0.05);生存分析证明高表达HNRNPA2B1的乳腺癌患者预后不良(P<0.05);双荧光素酶实验证明miR-204和HNRNPA2B1可靶向结合(P<0.05)。Western blot和Transwell实验证明,miR-204通过靶向调控HNRNPA2B1抑制乳腺癌的侵袭和迁移,且差异具有统计学意义(P<0.05)。结论 miR-204在乳腺癌组织和细胞中表达降低,且miR-204可通过靶向调控HNRNPA2B1抑制乳腺癌的侵袭和转移。
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MiR-204 inhibits invasion and metastasis of breast cancer cells by targeted regulation of
HNRNPA2B1 |
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Abstract: | Objective To investigate the effect of miR-204 on the invasion and metastasis of breast cancer by targeted regulation
of HNRNPA2B1. Methods The bioinformatics database was used to obtain data of the expressions of miR-204 in breast cancer
patients and the survival rate of the patients. RT-qPCR was used to detect the expression of miR-204 in breast cancer cell lines.
The expression vector GV369-miR-204 was used to overexpress miR-204 in MDA-MB-231 cells. Transwell assay was performed
to detect the effect of miR-204 on the migration and invasion ability of the breast cancer cells. The key genes (hub genes) of
miR-204 were determined by bioinformatics method. A dual luciferase assay was used to analyze the targeting relationship
between miR-204 and HNRNPA2B1. The expression of HNRNPA2B1 in MDA-MB-231 cells after miR-204 overexpression was
detected by Western blotting, and Transwell assay was used to examine the changes in the cell invasion ability. Results The
expression of miR-204 was decreased in both breast cancer tissues, and was significantly lower in breast cancer MDA-MB-231
cells than in MCF-10A cells (P<0.05). The decreased expression of miR-204 was associated with poorer prognosis of breast
cancer patients (P<0.05). Upregulation of miR-204 in MDA-MB-231 cells significantly inhibited the invasion and migration of
the cells (P<0.05). Analysis of the data from the Starbase revealed that the expression of miR-204-5p was negatively correlated
with the expression of HNRNPA2B1, and the expression
of HNRNPA2B1 was increased in breast cancer patients
(P<0.05) in association with a poorer prognosis of the
patients (P<0.05). Dual luciferase assay demonstrated
that miR-204 could bind to HNRNPA2B1 in a
target-specific manner. Western blotting and Transwell
assay showed that miR-204 significant inhibited the
migration and invasion ability of breast cancer cells by
targeting HNRNPA2B1 (P<0.05). Conclusion miR-204
expression is decreased in breast cancer tissues and cells,
and its overexpression can inhibit the invasion and
metastasis of breast cancer cells by targeted regulation of
HNRNPA2B1. |
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