| 梁猛.微小 RNA-525-5p通过靶向 RECQ样蛋白 5调控乳腺癌放射敏感性[J].安徽医药,2022,26(10):2032-2037. |
| 微小 RNA-525-5p通过靶向 RECQ样蛋白 5调控乳腺癌放射敏感性 |
| MicroRNA-525-5p regulates radiosensitivity of breast cancer by targeting RECQ protein-like 5 |
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| DOI：10.3969/j.issn.1009-6469.2022.10.029 |
| 中文关键词: 乳腺肿瘤 RecQ解旋酶类 微小 RNA-525-5p 放射敏感性 |
| 英文关键词: Breast neoplasms RecQ helicases miR-525-5p Radiosensitivity |
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| 中文摘要: |
| 目的探讨微小 RNA-525-5p(miR-525-5p)对乳腺癌细胞辐射照射敏感性的影响及机制。方法该研究起止时间为 2019年 6月至 2020年 6月，人乳腺癌细胞 MDA-MB-231、MCF-7、BT474、非恶性乳腺上皮细胞 MCF-10A均购自美国菌种保藏中心。运用 qRT-PCR法检测人乳腺癌细胞 MDA-MB-231、MCF-7、BT474、非恶性乳腺上皮细胞 MCF-10A中 miR-525-5p的 mRNA的表达；将 miR-525-5p模拟物( miR-525-5p mimics)阴性对照( miR-NC)组(转染 miR-NC)、 miR-525-5p组(转染 miR-525-5p mim. ics)、 RECQL5小干扰 RNA阴性对照( si-NC)组(转染 si-NC)、 RECQL5小干扰 RNA(si-RECQL5)组(转染 si-RECQL5)、 miR-525-5p+RECQL5过表达空载体( pcDNA)组(共转染 miR-525-5p mimics和 pcDNA)、 miR-525-5p+RECQL5过表达载体( pcDNA-REC. QL5)组(共转染 miR-525-5p mimics和 pcDNA-RECQL5)，均用脂质体法转染至 MDA-MB-231细胞； Western blotting检测细胞中 RECQ样蛋白 5(RECQL5)的蛋白表达；流式细胞术检测细胞凋亡；双荧光素酶报告基因检测实验检测细胞的荧光活性；克隆形成实验检测细胞的存活分数。结果与非恶性乳腺上皮细胞 MCF-10A相比，人乳腺癌细胞 MDA-MB-231、MCF-7、BT474中 miR-525-5p表达［ 0.28±0.02，0.33±0.02，0.42±0.03比 1.01±0.08］显著降低， RECQL5 mRNA和蛋白表达［ 1.68±0.15，1.58±0.12， |
| 英文摘要: |
| Objective To investigate the effect and mechanism of microRNA-525-5p (miR-525-5p) on the sensitivity of breast can. cer cells to radiation exposure.Methods The start and end time of this study was from June 2019 to June 2020. Human breast cancer cell MDA-MB-231, MCF-7, BT474 and non-malignant mammary epithelial cell MCF-10A were purchased from the American Culture Collection. qRT-PCR was used to detect the expressions of miR-525-5p mRNA in human breast cancer cell line MDA-MB-231, MCF-7, BT474 and non-malignant mammary epithelial cell MCF-10A; miR-525-5p mimics negative control (miR-NC) group (transfected miR-NC), the miR-525-5p group (transfected miR-525-5p mimics), RECQL5 small interfering RNA negative control (si-NC) group (transfected si-NC), RECQL5 small interfering RNA (si-RECQL5) group (transfected si -RECQL5), miR-525-5p+RECQL5 overexpres. sion empty vector (pcDNA) group (co-transfected miR-525-5p mimics and pcDNA), and miR-525-5p+RECQL5 overexpression vector (pcDNA-RECQL5) group (co-transfected miR-525-5p mimics and pcDNA-RECQL5) all transfected into MDA-MB-231 cells by lipo. some. The protein expression of RECQ protein-like 5 (RECQL5) in cells was detected by Western blotting, the apoptosis was detectedby flow cytometry, and the fluorescence activity of cells was detected by double luciferase reporter gene assay. The colony formation as.say detected the survival fraction of the cells.Results Compared with non-malignant mammary epithelial cells MCF-10A, the expres. sions of miR-525-5p [(0.28±0.02), (0.33±0.02), (0.42±0.03) vs. (1.01±0.08)] were significantly decreased and the mRNA and protein ex. pressions of RECQL5 [(1.68±0.15), (1.58±0.12), (1.48±0.11) vs. (1.00±0.08); (1.43±0.11), (1.38±0.12), (1.53±0.14) vs. (0.99±0.07)] were significantly increased in human breast cancer cells MDA-MB-231, MCF-7 and BT474(P<0.05). Overexpression of miR-525-5p or knockdown of RECQL5 could both promote the apoptosis of breast cancer cells MDA-MB-231 and enhance the sensitivity to radia. tion exposure. MiR-525-5p inhibited the fluorescence activity of wild-type RECQL5 cells and negatively regulated the expression ofRECQL5. Overexpression of RECQL5 reversed the apoptosis promotion and irradiation sensitization of breast cancer cells by miR-525-5p.Conclusion MiR-525-5p can promote the apoptosis of breast cancer cells and enhance the sensitivity to radiation exposure. Themechanism is related to the targeting of RECQL5, which will provide a new idea for radiation therapy of breast cancer. |
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