miR-339-5p调控EMT影响乳腺癌细胞化疗耐药的机制

doi:10.3760/cma.j.issn.1007-1245.2024.19.017

摘要/Abstract

摘要：

目的　分析miR-339-5p调控上皮细胞-间充质转化（EMT）影响乳腺癌细胞化疗耐药的机制。方法　于2023年2月至2024年2月体外培养正常乳腺上皮细胞系（MCF10A）及乳腺癌细胞系MCF-7，构建乳腺癌紫杉醇耐药细胞系MCF-7/紫杉醇，将MCF-7/紫杉醇细胞分别转染miR-339-5p模拟物（miR-339-5p mimics组）、miR-339-5p模拟物阴性对照（miR-NC组）、miR-339-5p抑制剂（miR-339-5p inhibitor组）及miR-339-5p抑制剂对照（NC组）。采用实时荧光定量逆转录聚合酶链反应（qRT-PCR）对miR-339-5p水平予以检测，以噻唑蓝（MTT）法测定细胞增殖抑制率，经流式细胞仪测定细胞凋亡情况，采用Western Blot法检测EMT相关蛋白［E-cadhesin（E-cad）、Vimentin（Vim）］的表达。采用独立样本t检验、ONE-WAY ANOVA分析及LSD-t检验对所获得的数据进行统计学分析。结果　miR-339-5p在MCF-7细胞的表达水平低于MCF10A细胞［（0.36±0.07）比（0.73±0.08），P<0.05］；转染48 h后，miR-339-5p mimics组细胞增殖抑制率、细胞凋亡率高于miR-NC组［（45.86±4.92）%比（20.44±2.16）%、（16.54±1.67）%比（4.23±0.45）%，均P<0.05］，miR-339-5p inhibitor组细胞增殖抑制率、细胞凋亡率低于NC组［（9.74±1.05）%比（17.18±1.84）%、（2.28±0.46）%比（5.43±0.59）%，均P<0.05］；miR-339-5p mimics组E-cad水平高于miR-NC组，Vim低于miR-NC组［（0.78±0.07）比（0.42±0.05）、（0.42±0.05）比（0.61±0.07），均P<0.05］，miR-339-5p inhibitor组E-cad低于NC组，Vim高于NC组［（0.23±0.04）比（0.34±0.05）、（0.84±0.09）比（0.69±0.08），均P<0.05］。结论　miR-339-5p可降低乳腺癌细胞化疗耐药性，抑制细胞增殖并促进细胞凋亡，其机制可能是通过调控EMT相关蛋白E-cad、Vim而抑制EMT。

关键词:

乳腺癌, miR-339-5p, 上皮细胞-间充质转化, 化疗耐药, 机制

Abstract:

Objective　To analyze the mechanism of miR-339-5p influencing chemotherapy resistance of breast cancer cells via regulating epithelial mesenchymal transformation (EMT). Methods　From February 2023 to February 2024, MCF10A mammary epithelial cells and MCF-7 breast cancer cell lines were cultured in vitro. The paclitaxel resistant breast cancer cell line (MCF-7/paclitaxel) was constructed. The MCF-7/paclitaxel cells were divided into a miR-339-5p mimics group, a miR-339-5p mimetic negative control group (miR-NC group), a miR-339-5p inhibitor group, and a miR-339-5p inhibitor control group (NC group). The real-time fluorescence quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expression level of miR-339-5p. The methyl thiazolyl tetrazolium (MTT) assay was used to determine the inhibition rate of cell proliferation. The flow cytometry was used to determine cell apoptosis. The expression of EMT related proteins [E-cadherin (E-cad) and Vimentin (Vim)] was detected using the Western blot method. The independent sample t test, one-way ANOVA, and LSD-t test were used for the statistical analysis of the obtained data. Results　The expression level of miR-339-5p in the MCF-7 cells was lower than that in the MCF10A cells [(0.36±0.07) vs. (0.73±0.08); P<0.05]. Forty-eight h after transfection, the proliferation inhibition rate and apoptosis rate in the miR-339-5p mimics group were higher than those in the miR-NC group [(45.86±4.92)% vs. (20.44±2.16)% and (16.54±1.67)% vs. (4.23±0.45)%; both P<0.05]. The cell proliferation inhibition rate and apoptosis rate of the miR-339-5p inhibitor group were lower than those of the NC group [(9.74±1.05)% vs.(17.18±1.84)% and (2.28±0.46)% vs. (5.43±0.59)%, both P<0.05]. The E-cad level in the miR-339-5p mimics group was higher than that in the miR-NC group [(0.78±0.07) vs. (0.42±0.05); P<0.05]; the Vim level in the miR-339-5p mimics group was lower than that in the miR-NC group [(0.42±0.05) vs. (0.61±0.07); P<0.05]. The E-cad level in the miR-339-5p inhibitor group was lower than that in the NC group [(0.23±0.04) vs. (0.34±0.05); P<0.05]; the Vim level in the miR-339-5p inhibitor group was higher than that in the NC group [(0.84±0.09) vs. (0.69±0.08); P<0.05]. Conclusions　MiR-339-5p can reduce chemotherapy resistance of breast cancer cells, inhibit cell proliferation, and promote cell apoptosis. The mechanism may be that it inhibits EMT regulating EMT related proteins——E-cad and Vim.

Key words:

Breast cancer, MiR-339-5p, Epithelial mesenchymal transformation, Chemotherapy resistance, Mechanism

李志路乔喜婷焦婉王小娟司小敏.

miR-339-5p调控EMT影响乳腺癌细胞化疗耐药的机制 [J]. 国际医药卫生导报, 2024, 30(19): 3250-3254.

Li Zhilu, Qiao Xiting, Jiao Wan, Wang Xiaojuan, Si Xiaomin.

Mechanism of miR-339-5p influencing chemotherapy resistance of breast cancer cells via regulating EMT [J]. International Medicine and Health Guidance News, 2024, 30(19): 3250-3254.

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