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miRNA-155 干扰对三阴性乳腺癌细胞作用的研究。

Research on the effect of interfering with miRNA-155 on triple-negative breast cancer cells.

机构信息

Department of Thyroid and Breast Diagnosis and Treatment Center, Shulan (Hangzhou) Hospital Affiliated to Zhejiang Shuren University Shulan International Medical College, Hangzhou, 310000, China.

Department of Geriatrics General Surgery, The First Affiliated Hospital of Kunming Medical University, Kunming, 650032, China.

出版信息

Genes Genomics. 2022 Sep;44(9):1117-1124. doi: 10.1007/s13258-021-01106-y. Epub 2021 Apr 28.

Abstract

BACKGROUND

Triple negative breast cancer (TNBC) is a poor prognosis breast cancer with the highest mutation rate and limited treatment options. MiR-155 is highly expressed in TNBC, but its role and potential mechanism in TNBC remain to be elucidated.

OBJECTIVE

The aim of this study is to examine the effect of interfering with miRNA-155 on the inflammatory pathway of NLRP 3 in TNBC (MDA-MB-231).

METHODS

MiRNA-155-specific interference (Si-miR-155) on MDA-MB-231 cell was manifested by transfection of miRNA-155 inhibitor. Meanwhile, blank control (Blank) and negative control (NC) were set. Cell growth and proliferation rate were detected by MTT; apoptosis rate were detected by flow cytometry; colony forming test was used to detected cell viability; cell migration ability was detected by Wound healing assay; TNF-α, IL-18, IL-6 and IL-1β levels were detected by ELISA. The mRNA of miRNA-155, NLRP3, ASC, caspase-1 and Ki67 were detected by qRT-PCR. The expression levels of NLRP3, caspase-1, ASC and Ki67 were detected by Western blotting.

RESULTS

The proliferation rate of Si-miRNA-155 group decreased, while the apoptosis rate increased significantly. After interfering with miRNA-155, the number of cancer cell colonies and the migration ability was decreased, and the secretion levels of IL-18, TNF-α, IL-6 and IL-1β were also inhibited. Moreover the mRNA and protein expression of NLRP3, caspase-1, ASC and Ki67 were significantly suppressed.

CONCLUSIONS

Interference with miRNA-155 can inhibit the NLRP3 pathway of MDA-MB-231 cells, as well as the proliferation, migration and inflammatory factor secretion of MDA-MB-231 cell, and can accelerate its apoptosis.

摘要

背景

三阴性乳腺癌(TNBC)是一种预后不良的乳腺癌,其突变率最高,治疗选择有限。miR-155 在 TNBC 中高度表达,但它在 TNBC 中的作用和潜在机制仍有待阐明。

目的

本研究旨在研究干扰 miRNA-155 对 TNBC(MDA-MB-231)中 NLRP3 炎症途径的影响。

方法

通过转染 miRNA-155 抑制剂来表现 MDA-MB-231 细胞中的 miRNA-155 特异性干扰(Si-miR-155)。同时,设置空白对照(Blank)和阴性对照(NC)。通过 MTT 检测细胞生长和增殖率;通过流式细胞术检测细胞凋亡率;通过集落形成试验检测细胞活力;通过划痕愈合试验检测细胞迁移能力;通过 ELISA 检测 TNF-α、IL-18、IL-6 和 IL-1β 水平。通过 qRT-PCR 检测 miRNA-155、NLRP3、ASC、caspase-1 和 Ki67 的 mRNA。通过 Western blot 检测 NLRP3、caspase-1、ASC 和 Ki67 的表达水平。

结果

Si-miRNA-155 组的增殖率降低,而凋亡率显著增加。干扰 miRNA-155 后,癌细胞集落数和迁移能力降低,IL-18、TNF-α、IL-6 和 IL-1β 的分泌水平也受到抑制。此外,NLRP3、caspase-1、ASC 和 Ki67 的 mRNA 和蛋白表达均显著受抑制。

结论

干扰 miRNA-155 可抑制 MDA-MB-231 细胞的 NLRP3 途径,以及 MDA-MB-231 细胞的增殖、迁移和炎症因子分泌,并可加速其凋亡。

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