miRNA-21对乳腺癌细胞放射敏感性的影响及其机制

doi:10.13481/j.1671-587x.20190405

吉林大学学报(医学版) ›› 2019, Vol. 45 ›› Issue (04): 772-778.doi: 10.13481/j.1671-587x.20190405

• 基础研究 • 上一篇

miRNA-21对乳腺癌细胞放射敏感性的影响及其机制

信瑞¹, 李冬², 徐慧英³, 王春宇⁴, 曲丹华⁵, 孙海峰¹

1. 吉林大学第二医院放射线科, 吉林长春 130041;
2. 吉林大学第二医院妇产科, 吉林长春 130041;
3. 吉林大学第一医院联合超声科, 吉林长春 130021;
4. 吉林大学第二医院放疗科, 吉林长春 130041;
5. 吉林大学第二医院呼吸与危重症医学科, 吉林长春 130041

收稿日期:2018-12-04 发布日期:2019-08-02
通讯作者: 曲丹华,副主任护师(Tel:0431-81136487,E-mail:61505502@qq.com);孙海峰,主管技师(Tel:0431-81136613,E-mail:53518525@qq.com) E-mail:61505502@qq.com;53518525@qq.com
作者简介:信瑞(1982-),男,黑龙江省齐齐哈尔市人,副主任技师,医学博士,主要从事放射生物学方面的研究。
基金资助:
吉林省科技厅优秀青年人才基金资助课题（20180520127JH）

Influence of miRNA-21 in radiosensitivity of breast cancer cells and its mechanism

XIN Rui¹, LI Dong², XU Huiying³, WANG Chunyu⁴, QU Danhua⁵, SUN Haifeng¹

1. Department of Radiology, Second Hospital, Jilin University, Changchun 130041, China;
2. Department of Obstetrics and Gynecology, Second Hospital, Jilin University, Changchun 130041, China;
3. Department of Ultrasonography, First Hospital, Jilin University, Changchun 130021, China;
4. Department of Radiotherapy, Second Hospital, Jilin University, Changchun 130041, China;
5. Department of Respiration and Critical Disease, Second Hospital, Jilin University, Changchun 130041, China

Received:2018-12-04 Published:2019-08-02

摘要/Abstract

摘要： 目的：观察miRNA-21对乳腺癌细胞放射敏感性的影响，并初步探讨其作用机制。方法：乳腺癌T47D和MDA-MB-361细胞分别给予0.0、2.5和5.0 Gy γ射线照射，CCK8法检测细胞存活率，流式细胞术分析细胞周期进程，实时定量PCR法检测72 h内细胞中miRNA-21表达水平。T47D和MDA-MB-361细胞分别转染anti-miRNA-21序列（anti-miRNA-21组）和阴性对照序列（阴性对照组），并设置未转染细胞为空白对照组，实时定量PCR法检测各组细胞中miRNA-21表达水平；经0.0和5.0 Gy γ射线照射后，CCK8法检测细胞存活率，流式细胞术分析细胞周期进程。结果：经5.0 Gy γ射线照射后，T47D细胞存活率明显高于MDA-MB-361细胞（P<0.05）；与0.0 Gy照射组比较，5.0 Gy照射组T47D和MDA-MB-361细胞G₂/M期细胞百分率均明显升高（P<0.05），miRNA-21表达水平均明显升高（P<0.05）。与空白对照组比较，anti-miRNA-21组T47D和MDA-MB-361细胞中miRNA-21表达水平均明显降低（P<0.05或P<0.01）；给予5.0 Gy γ射线照射后，anti-miRNA-21组T47D和MDA-MB-361细胞存活率均明显降低（P<0.05或P<0.01），T47D细胞G₂/M期细胞百分率明显降低（P<0.05），而subG₁期细胞百分率明显升高（P<0.05）。结论：miRNA-21表达下调能够增强乳腺癌细胞的放射敏感性，其机制可能与抑制细胞周期G₂/M期阻滞有关。

关键词: miRNA-21, 乳腺肿瘤, 放射敏感性, 细胞存活率, G₂/M期阻滞

Abstract: Objective:To observe the influence of miRNA-21 in the radiosensitivity of the breast cancer cells, and to preliminarily explore its mechanism. Methods:The breast cancer T47D and MDA-MB-361 cells were irradiated with 0.0,2.5,and 5.0 Gy γ-ray,respectively; CCK8 assay was used to detect the survival rates of T47D and MDA-MB-361 cells and flow cytometry was used to analyze the cell cycle; real-time quantitative PCR was used to detect the expression levels of miRNA-21 in cells during 72 h. The T47D and MDA-MB-361 cells were transfected with anti-miRNA-21 sequence (anti-miRNA-21 group) and negative control sequence(negative control group), respectively; the untransfected cells were set as blank control group. Real-time quantitative PCR was used to detect the expression levels of miRNA-21 in cells in various groups. After irradiation with 0.0 and 5.0 Gy γ-ray, CCK8 assay was used to detect the survival rates, and flow cytometry was used to analyze the cell cycle. Results:After irradiation with 5.0 Gy γ-ray, the survival rate of T47D cells was significantly higher than that of the MDA-MB-361 cells (P<0.05). Compared with 0.0 Gy radiation group, the percentages of T47D and MDA-MB-361 cells in G₂/M phase in 5.0 Gy radiation group were significantly increased(P<0.05); the miRNA-21 expression levels in the T47D and MDA-MB-361 cells were significantly increased (P<0.05).The miRNA-21 expression levels in the T47D and MDA-MB-361 cells in anti-miRNA-21 group were lower than those in blank control group (P<0.05). After irradiation with 5.0 Gy γ-ray, the survival rates of T47D and MDA-MB-361 cells in anti-miRNA-21 group were significantly lower than those in blank control group(P<0.05 or P<0.01), and the percentage of T47D cells in G₂/M phase in anti-miRNA-21 group was significantly lower than that in blank control group (P<0.05), and the percentage of T47D cells in subG₁ phase was significantly higher than that in blank control group (P<0.05). Conclusion:The down-regulation of miRNA-21 may enhance the radiosensitivity of breast cancer cells, and its mechanism may be related to the inhibition of G₂/M phase arrest.

Key words: miRNA-21, breast neoplasms, radiosensitivity, cell survival rate, G₂/M phase arrest

中图分类号:

R737.9

信瑞, 李冬, 徐慧英, 王春宇, 曲丹华, 孙海峰. miRNA-21对乳腺癌细胞放射敏感性的影响及其机制[J]. 吉林大学学报(医学版), 2019, 45(04): 772-778.

XIN Rui, LI Dong, XU Huiying, WANG Chunyu, QU Danhua, SUN Haifeng. Influence of miRNA-21 in radiosensitivity of breast cancer cells and its mechanism[J]. Journal of Jilin University(Medicine Edition), 2019, 45(04): 772-778.

参考文献

[1] 惠周光, 张烨, 张江鹄, 等. 2010年与2004年中国大陆地区乳腺癌改良根治术后放疗现状比较[J].中华放射肿瘤学杂志, 2012, 21(4):352-356.
[2] CROKER A K, ALLAN A L. Inhibition of aldehyde dehydrogenase (ALDH) activity reduces chemotherapy and radiation resistance of stem-like ALDHhiCD44⁺ human breast cancer cells[J]. Breast Cancer Res Treat, 2012, 133(1):75-87.
[3] KUMAR S, KEERTHANA R, PAZHANIMUTHU A, et al. Overexpression of circulating miRNA-21 and miRNA-146a in plasma samples of breast cancer patients[J]. Indian J Biochem Biophys, 2013, 50(3):210-214.
[4] ZHANG H L, YANG L F, ZHU Y, et al. Serum miRNA-21:Elevated levels in patients with metastatic hormone-refractory prostate cancer and potential predictive factor for the efficacy of docetaxel-based chemotherapy[J]. Prostate, 2011, 71(3):326-331.
[5] COTE G A, GORE A J, MCELYEA S D, et al. A pilot study to develop a diagnostic test for pancreatic ductal adenocarcinoma based on differential expression of select miRNA in plasma and bile[J]. Am J Gastroenterol, 2014, 109(12):1942-1952.
[6] BARANWAL S, ALAHARI S K. miRNA control of tumor cell invasion and metastasis[J]. Int J Cancer, 2010, 126(6):1283-1290.
[7] WANG X C,WANG W,ZHANG Z B, et al. Overexpression of miRNA-21 promotes radiation-resistance of non-small cell lung cancer[J]. Radiat Oncol, 2013, 8:146.
[8] WONG S T, ZHANG X Q, ZHUANG J T, et al. MicroRNA-21 inhibition enhances in vitro chemosensitivity of temozolomide-resistant glioblastoma cells[J]. Anticancer Res, 2012, 32(7):2835-2841.
[9] KUMARSWAMY R, VOLKMANN I, THUM T. Regulation and function of miRNA-21 in health and disease[J]. RNA Biol, 2011, 8(5):706-713.
[10] 钟莉莉, 赵银龙, 李炳锦, 等. MicroRNA-210对卵巢癌细胞生长及放射敏感性的影响[J]. 吉林大学学报:医学版, 2018, 44(2):265-269.
[11] MARTIN S L, KALA R, TOLLEFSBOL T O. Mechanisms for the inhibition of colon cancer cells by sulforaphane through epigenetic modulation of MicroRNA-21 and human telomerase reverse transcriptase (hTERT) down-regulation[J]. Curr Cancer Drug Targets, 2018, 18(1):97-106.
[12] KUMAR S, KEERTHANA R, PAZHANIMUTHU A, et al. Overexpression of circulating miRNA-21 and miRNA-146a in plasma samples of breast cancer patients[J]. Indian J Biochem Biophys, 2013, 50(3):210-214.
[13] SHENOUDA S K, ALAHARI S K. MicroRNA function in cancer:oncogene or a tumor suppressor?[J]. Cancer Metastasis Rev, 2009, 28(3/4):369-378.
[14] WANG Z X, LU B B, WANG H, et al. MicroRNA-21 modulates chemosensitivity of breast cancer cells to doxorubicin by targeting PTEN[J]. Arch Med Res, 2011, 42(4):281-290.
[15] 狄英波, 孙颖, 张桂英, 等. MicroRNA-21与乳腺癌细胞MDA-MB-231放射敏感性之间关系的研究[J]. 中国妇幼保健, 2012, 27(34):5571-5573.
[16] GABRIELY G, WURDINGER T, KESARI S, et al. MicroRNA 21 promotes glioma invasion by targeting matrix metalloproteinase regulators[J]. Mol Cell Biol, 2008, 28(17):5369-5380.
[17] CARLETTI M Z, FIEDLER S D, CHRISTENSON L K. MicroRNA 21 blocks apoptosis in mouse periovulatory granulosa cells[J]. Biol Reprod, 2010, 83(2):286-295.
[18] ZHANG L, KANG W Q, LU X L, et al. LncRNA CASC11 promoted gastric cancer cell proliferation, migration and invasion in vitro by regulating cell cycle pathway[J]. Cell Cycle, 2018, 17(15):1886-1900.
[19] ZHANG W F, XIONG Y W, ZHU T T, et al. MicroRNA let-7g inhibited hypoxia-induced proliferation of PASMCs via G₀/G₁ cell cycle arrest by targeting c-myc[J]. Life Sci, 2017, 170:9-15.
[20] ANASTASOV N, HÖFIG I, VASCONCELLOS I G, et al. Radiation resistance due to high expression of miR-21 and G₂/M checkpoint arrest in breast cancer cells[J]. Radiat Oncol, 2012, 7:206.

miRNA-21对乳腺癌细胞放射敏感性的影响及其机制

Influence of miRNA-21 in radiosensitivity of breast cancer cells and its mechanism

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