[1]王亮,黄琰菁,陈晶,等.miR-3942-3p负调控靶基因BARD1增加子宫内膜癌[J].徐州医科大学学报,2021,41(03):166-172.[doi:10.3969/j.issn.2096-3882.2021.03.003]
 MiR-3942-3p increases the radiosensitivity of endometrial[J].Journal of Xuzhou Medical University,2021,41(03):166-172.[doi:10.3969/j.issn.2096-3882.2021.03.003]
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miR-3942-3p负调控靶基因BARD1增加子宫内膜癌()
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《徐州医科大学学报》[ISSN:2096-3882/CN:32-1875/R]

卷:
41
期数:
2021年03期
页码:
166-172
栏目:
出版日期:
2021-03-25

文章信息/Info

Title:
MiR-3942-3p increases the radiosensitivity of endometrial
作者:
王亮 黄琰菁 陈晶 王天柱
海南省肿瘤医院放疗科,海南海口570311
关键词:
miR-3942-3pBARD1子宫内膜癌放射敏感性
分类号:
R737.33
DOI:
10.3969/j.issn.2096-3882.2021.03.003
文献标志码:
A
摘要:
目的探讨微小RNA-132-3p(microRNA-132-3p,miR-3942-3p)对子宫内膜癌细胞放射敏感性的影响及其可能的作用机制。方法实时荧光定量聚合酶链反应(quantitative real-time PCR,RT-qPCR)检测子宫内膜癌细胞(HEC-1A细胞)和人子宫内膜上皮细胞(HEEC细胞)中miR-3942-3p和BRCA1关联环域1(BRCA1 associated RING domain 1,BARD1)的表达量。X射线(0、2、4、6 Gy)照射HEC-1A细胞后,RT-qPCR检测miR-3942-3p和BARD1的表达量。下调miR-3942-3p表达且经6 Gy X射线照射后,分别利用CCK-8、流式细胞仪和蛋白质免疫印迹(Western blot)检测HEC-1A细胞增殖能力、细胞凋亡率、B淋巴细胞瘤-2基因(B-cell lymphoma-2,Bcl-2)和Bcl-2-相关X蛋白(BCL2-Associated X,Bax)表达量。Targetscan和双荧光素报告基因实验分析miR-3942-3p与BARD1之间的关系。下调BARD1表达且经6 Gy X射线照射后,分析HEC-1A细胞增殖能力、细胞凋亡率以及Bax、Bcl-2表达量。检测miR-3942-3p通过BARD1对HEC-1A细胞放射敏感性的影响。结果和癌旁正常组织相比,子宫内膜癌组织中miR-3942-3p的表达量明显降低(P<0.05),BARD1表达量明显升高(P<0.05)。和HEEC细胞相比,HEC-1A细胞中miR-3942-3p的表达量明显降低(P<0.01),BARD1表达量明显升高(P<0.01)。miR-3942-3p表达量随着X射线放射剂量增加而上升,BARD1表达量随着X射线放射剂量增加而降低。HEC-1A细胞经6 Gy X射线照射后,下调miR-3942-3p降低了HEC-1A细胞的放射敏感性。miR-3942-3p靶向BARD1。下调BARD1后经6 Gy X射线照射显著提高HEC-1A细胞增殖能力,降低细胞凋亡率,上调miR-3942-3p通过BARD1增强了HEC-1A细胞的放射敏感性。结论上调miR-3942-3p负调控靶基因BARD1增强子宫内膜癌细胞的放射敏感性。
Abstract:
ObjectiveTo investigate the effects of miR-3942-3p on the radiosensitivity of endometrial carcinoma cells and its possible mechanism.MethodsThe levels of miR-3942-3p and BRCA1 associated RING domain 1 (BARD1) in human endometrial cancer -1A (HEC-1A) cells and human endometrial epithelial cells(HEECs) were detected by quantitative real-time PCR (RT-qPCR). HEC-1A cells were irradiated with 0, 2, 4 and 6 Gy of X-rays. The amounts of miR-3942-3p and BARD1 were detected by RT-qPCR.After down-regulating the expression of miR-3942-3p and 6 Gy X-ray irradiation, the proliferation and apoptosis of HEC-1A cells were detected by CCK-8 assay and flow cytometry, respectively; while the amounts of B-cell lymphoma-2 (Bcl-2) and BCL2-associated X (Bax) in HEC-1A cells were measured by Western blot. The relationship between miR-3942-3p and BARD1 was analyzed by Targetscan and dual luciferin reporter assay. After down-regulating BARD1 expression and 6 Gy X ray irradiation, the proliferation and apoptosis of HEC-1A cells were detected by CCK-8 assay and flow cytometry, respectively, while the amounts of Bcl-2 and Bax in HEC-1A cells were measured by Western blot. The effects of miR-3942-3p via BARD1 on the radiosensitivity of HEC-1A cells were detected. ResultsCompared with those in adjacent non-cancer tissues, the levels of miR-3942-3p in endometrial carcinoma tissues significantly decreased (P<0.05),but the amounts of BARD1 significantly increased (P<0.05). Compared with HEEC cells, the levels of miR-3942-3p in HEC-1A cells significantly decreased (P<0.01), but the amounts of BARD1 significantly increased (P<0.01). The expression of miR-3942-3p increased after radiation in a dose-dependent manner. The expression of BARD1 decreased after radiation in a dose-dependent manner. After 6 Gy X-ray irradiation,down-regulation of miR-3942-3p reduced the radiosensitivity of HEC-1A cells. MiR-3942-3p targeted on BARD1. After downregulation of BARD1, 6 Gy X-ray irradiation increased the proliferation abilities of HEC-1A cells and reduced the apoptotic rate. Upregulation of miR-3942-3p enhanced the radiosensitivity of HEC-1A cells via BARD1.ConclusionsUp-regulation of miR-3942-3p negatively targets BARD1 and enhance the radiosensitivity of endometrial carcinoma cells.
更新日期/Last Update: 2021-04-25