微小RNA-152对乳腺癌细胞增殖及阿霉素敏感性影响的实验研究

临床肿瘤学杂志 ›› 2018, Vol. 23 ›› Issue (5): 391-396.

微小RNA-152对乳腺癌细胞增殖及阿霉素敏感性影响的实验研究

473000 河南南阳南阳医学高等专科学校第一附属医院肿瘤内科

收稿日期:2018-01-22 修回日期:2018-03-16 出版日期:2018-05-31 发布日期:2018-06-07

Effect of microRNA-152 on the proliferation of breast cancer cells and the sensitivity to adriamycin

Department of Oncology， the First Affiliated Hospital of Nanyang Medical College， Nanyang 473000， China

Received:2018-01-22 Revised:2018-03-16 Online:2018-05-31 Published:2018-06-07

摘要/Abstract

摘要： 目的探讨微小RNA-152（miR-152）对乳腺癌细胞阿霉素敏感性的影响及其作用机制。方法采用实时定量PCR（QPCR）检测人乳腺上皮细胞株HBL-100、人乳腺癌MCF-7细胞及阿霉素耐药株MCF-7／ADR细胞中的miR-152水平，分别向MCF-7细胞和MCF-7／ADR细胞转染miR-152模拟物 mimics（过表达组）和阴性对照NC（阴性对照组），以未行转染为空白对照组。采用QPCR检测转染后MCF-7细胞和MCF-7／ADR细胞中miR-152的表达情况，采用MTT法检测不同浓度阿霉素（10、50、100、200和500 ng/ml）处理后各组的增殖情况，流式细胞术检测转染后各组的凋亡率，Western blotting检测磷脂酰肌醇激酶-3催化亚基α基因（PIK3CA）的蛋白水平，双荧光素酶报告实验评价miR-152对PIK3CA的靶向调控作用。结果 QPCR实验结果表明MCF-7细胞和MCF-7／ADR细胞中的miR-152水平均低于HBL-100细胞，且MCF-7／ADR细胞的miR-152水平均低于MCF-7细胞，差异有统计学意义（P＜0.05）；过表达组转染后的miR-152水平均高于阴性对照组和空白对照组（P＜0.05）。与其余两组比较，过表达组的存活率降低而凋亡率升高，且PIK3CA蛋白水平也降低，差异有统计学意义（P＜0.05）。miR-152可抑制野生型PIK3CA 3’ UTR报告基因载体的荧光素酶活性，而对突变型PIK3CA 3’UTR的荧光素酶活性无影响。结论 miR-152表达在乳腺癌细胞中降低，且与阿霉素耐药有关，参与乳腺癌细胞的增殖和凋亡过程，在逆转乳腺癌耐药中发挥重要作用，具有一定价值。

关键词: 乳腺癌, 微小RNA-152, 磷脂酰肌醇激酶-3催化亚基α基因（PIK3CA）, 阿霉素

Abstract:

Objective To investigate the effect of miRNA-152 （miR-152） on adriamycin sensitivity of breast cancer cells and its mechanism. Methods The levels of miR-152 in human breast epithelial cell line HBL-100， human breast cancer MCF-7 cells and adriamycin resistant MCF-7／ADR cells were detected by real-time quantitative PCR （QPCR）. MCF-7 and MCF-7／ADR cells were transfected with miR-152 mimics （overexpression group） or negative control （negative control group）， and no transfection was used as blank control group. The expression of miR-152 in MCF-7 cells and MCF-7／ADR cells after transfection were detected by QPCR. The proliferation of cells treated with different concentrations of adriamycin（10, 50, 100, 200 and 500 ng/ml） was detected by MTT， and the apoptotic rate after transfection was detected by flow cytometry. Western blotting was used to detect the protein level of phosphatidylinositol-3-kinase catalytic subunit α （PIK3CA）. The target regulation of miR-152 on PIK3CA was evaluated by the double luciferase report test. Results QPCR Results showed that miR-152 levels in MCF-7 cells and MCF-7／ADR cells were all lower than those in HBL-100 cells， and miR-152 levels in MCF-7／ADR cells were lower than that in MCF-7 cells （P＜0.05）. The levels of miR-152 after transfection in overexpression group were higher than those in the negative control group and the blank control group （P＜0.05）. Compared with other two groups， the survival rate and the PIK3CA protein level of the overexpression group decreased and the apoptotic rate increased （P＜0.05）. MiR-152 could inhibite the luciferase activity of the wild-type PIK3CA 3’UTR reporter gene， but had no effect on the luciferase activity of the mutant PIK3CA 3’UTR. Conclusion The expression of miR-152 is decreased in breast cancer cells， and is related to adriamycin resistance in breast cancer cells. It is involved in the proliferation and apoptosis process of breast cancer cells， and plays an important role in reversing the drug resistance of breast cancer.

Key words: Breast cancer, MicroRNA-152, Phosphatidylinositol-3-kinase catalytic subunit α（PIK3CA）, Adriamycin

中图分类号:

R737.9

杨秀丽, 沈玲.

微小RNA-152对乳腺癌细胞增殖及阿霉素敏感性影响的实验研究 [J]. 临床肿瘤学杂志, 2018, 23(5): 391-396.

YANG Xiuli, SHEN Ling..

Effect of microRNA-152 on the proliferation of breast cancer cells and the sensitivity to adriamycin [J]. Chinese Clinical Oncology, 2018, 23(5): 391-396.

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