| 郭 猛,王 阳,何庆泗,刘道同,刘秀丽.MicroRNA-221/222对乳腺癌MDA-MB-231/DOX细胞阿霉素耐药性的影响[J].现代生物医学进展英文版,2020,(15):2843-2847. |
| MicroRNA-221/222对乳腺癌MDA-MB-231/DOX细胞阿霉素耐药性的影响 |
| Effect of Microrna-221/222 on Adriamycin Resistance of Breast Cancer MDA-MB-231/DOX Cells |
| Received:December 23, 2019 Revised:January 18, 2020 |
| DOI：10.13241/j.cnki.pmb.2020.15.008 |
| 中文关键词: MicroRNA-221/222 阿霉素 耐药性 乳腺癌 MDA-MB-231/DOX |
| 英文关键词: MicroRNA-221/222 Doxorubicin Drug resistance Breast cancer MDA-MB-231 /DOX |
| 基金项目:山东省自然科学基金项目（ZR2016YL639） |
| Author Name | Affiliation | E-mail | | GUO Meng | Department of Breast and Thyroid Surgery, Jining First People's Hospital Affiliated to Shandong University of Traditional Chinese Medicine, Jining, Shandong, 272111, China | qingsih@yeah.net | | WANG Yang | Department of Breast and Thyroid Surgery, Jining First People's Hospital Affiliated to Shandong University of Traditional Chinese Medicine, Jining, Shandong, 272111, China | | | HE Qing-si | Department of General Surgery, Qilu Hospital of Shandong University, Ji'nan, Shandong, 250012, China | | | LIU Dao-tong | Department of Breast and Thyroid Surgery, Jining First People's Hospital Affiliated to Shandong University of Traditional Chinese Medicine, Jining, Shandong, 272111, China | | | LIU Xiu-li | Department of Breast and Thyroid Surgery, Jining First People's Hospital Affiliated to Shandong University of Traditional Chinese Medicine, Jining, Shandong, 272111, China | |
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| 中文摘要: |
| 摘要 目的：探讨微小RNA-221/222（miR-221/222）对乳腺癌MDA-MB-231/阿霉素（DOX）细胞DOX耐药性的影响。方法：采用脂质体法转染miR-221/222抑制物（miR-221/222 inhibitor）至MDA-MB-231/DOX细胞内（Inhibitor组），同时设立空白对照组和转染无关序列的阴性对照组，采用实时荧光定量PCR（qRT-PCR）检测MDA-MB-231细胞株及MDA-MB-231/DOX细胞株的miR-221/222表达水平及转染效率；CCK-8法检测转染48 h后MDA-MB-231/DOX细胞对DOX药物敏感性的变化；流式细胞术（FCM）检测转染MDA-MB-231/DOX细胞的细胞凋亡率；蛋白免疫印迹实验（WB）检测转染后MDA-MB-231/DOX细胞内促凋亡蛋白p53上调凋亡调控因子（PUMA），Bcl2蛋白修饰因子（BMF）以及细胞周期蛋白激酶抑制因子p27（p27 Kip1）的表达情况。结果：MDA-MB-231/DOX细胞中的miR-221/222表达水平高于亲本MDA-MB-231细胞（P<0.05）；MDA-MB-231/DOX细胞转染miR-221/222 inhibitor 96 h后，miR-221/222的表达水平低于空白对照组和阴性对照组（P<0.05）；与空白对照组相比，MDA-MB-231/DOX细胞转染miR-221/222 inhibitor 48h后，DOX继续处理48 h后，细胞的凋亡率明显升高，且细胞内的促凋亡蛋白PUMA，BMF以及p27 Kip1的表达均增加（P<0.05）；DOX对inhibitor组耐药细胞的半数抑制浓度（IC50）显著低于空白对照组细胞及阴性对照组（P<0.05）。结论：miR-221/222能够增加MDA-MB-231/DOX细胞对DOX的耐药性，这可能与下调促凋亡蛋白的表达有关；降低miR-221/222水平可诱导MDA-MB-231/DOX凋亡，并且上调促凋亡蛋白的表达，从而部分逆转MDA-MB-231/DOX对DOX的耐药性。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of microrna-221/222 (miR-221/222) on DOX resistance of breast cancer MDA-MB-231/doxorubicin (DOX) cells. Methods: miR-221/222 inhibitor (miR-221/222 inhibitor) was transfected into MDA-MB-231/DOX cells by liposome method, the blank control group and the negative control group were set up meanwhile. The transfection efficiency and expression level of miR-221/222 in MDA-MB-231 cell line and cell line MDA-MB-231/DOX cell line was detected by real time fluorescent quantitative PCR (qRT-PCR); the change of drug sensitivity of MDA-MB-231/DOX cells to dox after 48 hours of transfection was detected by CCK-8 tests; the apoptosis rate of MDA-MB-231/DOX cells was detected by flow cytometry(FCM); the upregulation of pro-apoptotic protein p53 up regulates apoptosis (PUMA), bcl-2 protein modifying factor (BMF) and cyclin kinase inhibitor p27 (p27 kip1) in MDA-MB-231/DOX cells were detected by Western blotting (WB). Results: The expression level of miR-221/222 in MDA-MB-231/DOX cells was higher than that in MDA-MB-231 cells (P<0.05), the expression level of miR-221/222 in miR-221/222 inhibitor transfected by MDA-MB-231/DOX cells was lower than that in the blank control group and the negative control group (P<0.05) after 96 hours of treatment; compared with the blank control group, the expression level of miR-221/222 inhibitor transfected by MDA-MB-231/DOX cells was lower than that in the blank control group (P<0.05). The apoptotic rate of the cells increased significantly and the expression of apoptosis promoting protein puma, BMF and p27 kip1 in the cells increased (P<0.05); the 50% inhibitory concentration(IC50) of drug resistance cells in the inhibitor group was significantly lower than that in the blank control group and the negative control group (P<0.05). Conclusion: miR-221/222 can increase the resistance of MDA-MB-231/DOX cells to DOX, which may be related to down regulating the expression of pro-apoptotic protein, and down regulating the level of miR-221/222 can induce MDA-MB-231/DOX apoptosis and raise the expression of pro-apoptotic protein, which can partially reverse the resistance of MDA-MB-231/DOX to DOX. |
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