| miR-762对子宫内膜癌Ishikawa细胞放疗敏感性的影响及机制研究 |
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| 引用本文： | 李儒彩,潘英连,蔡兴锐,郭丽娟.miR-762对子宫内膜癌Ishikawa细胞放疗敏感性的影响及机制研究[J].现代肿瘤医学,2023,0(10):1805-1810. |
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| 作者姓名： | 李儒彩 潘英连 蔡兴锐 郭丽娟 |
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| 作者单位： | 1.海南医学院第一附属医院放疗科;2.肿瘤内科，海南 海口 570102 |
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| 基金项目： | 海南省卫生健康行业科研项目（编号:20A200431） |
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| 摘 要： | 目的:分析miR-762在子宫内膜癌（EC）患者癌组织中的表达水平及其对子宫内膜癌Ishikawa细胞放疗敏感性的影响及机制研究。方法:qRT-PCR检测54例EC癌组织和癌旁组织以及正常子宫内膜细胞ESC和EC细胞系中miR-762及腺瘤性结肠息肉2（APC2） mRNA表达水平。不同放射剂量处理Ishikawa细胞，qRT-PCR检测miR-762和APC2 mRNA表达水平。将Ishikawa细胞分为空白对照组（blank组）、Lv-NC组（感染miR-762 sponge阴性对照慢病毒），Lv-miR-762 SP组（感染miR-762 sponge慢病毒）、Lv-miR-762 SP+si-NC组和Lv-miR-762 SP+si-APC2组。qRT-PCR和Western blot检测miR-762和APC2表达水平。双荧光素酶报告基因实验检测miR-762和APC2的靶向关系。采用6 Gy射线照射各组Ishikawa细胞，克隆形成实验检测细胞放射敏感性;CCK-8检测细胞增殖活性;流式细胞术检测细胞凋亡;Western blot检测细胞中CyclinD1、p21、Bcl-2和Bax等蛋白表达水平。结果:在EC癌组织及细胞系中miR-762高表达，而APC2低表达。随着射线照射剂量的增加，Ishikawa细胞中miR-762表达水平逐渐升高（P＜0.05），APC2 mRNA表达水平逐渐降低（P＜0.05），呈剂量依赖性。双荧光素酶报告基因实验证实，APC2是miR-762靶基因。抑制miR-762可明显下调miR-762表达水平，上调APC2表达水平，增强细胞放射敏感性，抑制细胞增殖，促进细胞凋亡，并下调CyclinD1和Bcl-2蛋白表达水平，上调p21和Bax蛋白表达水平（P＜0.05）。此外，干扰APC2基因可显著逆转抑制miR-762表达对Ishikawa细胞放射敏感性的促进作用。结论:miR-762在EC癌组织及细胞系中高表达，抑制其表达可通过上调APC2表达从而增强Ishikawa细胞的放射敏感性。
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| 关 键 词： | 子宫内膜癌 miR-762 APC2 增殖 凋亡 放射敏感性 |
Effects of miR-762 on radiosensitivity of endometrial carcinoma Ishikawa cells and its mechanism |
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| LI Rucai,PAN Yinglian,CAI Xingrui,GUO Lijuan.Effects of miR-762 on radiosensitivity of endometrial carcinoma Ishikawa cells and its mechanism[J].Journal of Modern Oncology,2023,0(10):1805-1810. |
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| Authors: | LI Rucai PAN Yinglian CAI Xingrui GUO Lijuan |
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| Affiliation: | 1.Department of Radiotherapy;2.Department of Medical Oncology,the First Affiliated Hospital of Hainan Medical University,Hainan Haikou 570102,China. |
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| Abstract: | Objective:To investigate the expression level of miR-762 in endometrial cancer (EC) patients' cancer tissues and its effects on radiosensitivity of endometrial cancer Ishikawa cells and its mechanism.Methods:qRT-PCR was used to detect the expression levels of miR-762 and adenomatous colonic polyp 2 (APC2) mRNA in 54 EC cancer tissues and adjacent tissues,as well as normal endometrial cells ESC and EC cell lines.Ishikawa cells were treated with different radiation doses,and the expression levels of miR-762 and APC2 mRNA were detected by qRT-PCR.Ishikawa cells were divided into blank control group (blank group),Lv-NC group (infected with miR-762 sponge negative control lentivirus),Lv-miR-762 SP group (infected with miR-762 sponge lentivirus),Lv-miR-762 SP+si-NC group and Lv-miR-762 SP+si-APC2 group.The expression levels of miR-762 and APC2 in cells were detected by qRT-PCR and Western blot.The targeting relationship between miR-762 and APC2 was detected by dual-luciferase reporter gene assay.Ishikawa cells in each group were irradiated with 6 Gy rays.The radiosensitivity of cells was detected by colony formation assay.Cell proliferation activity was detected by CCK-8.Cell apoptosis was detected by flow cytometry.The protein expression levels of CyclinD1,p21,Bcl-2 and Bax were detected by Western blot.Results:miR-762 was highly expressed in EC cancer tissues and cell lines,while APC2 was low expressed.With the increase of radiation dose,the expression level of miR-762 in Ishikawa cells was gradually increased (P＜0.05),and the expression level of APC2 mRNA was gradually decreased (P＜0.05),in a dose-dependent manner.The dual-luciferase reporter gene assay confirmed that APC2 was a target gene of miR-762.Inhibition of miR-762 could significantly down-regulate the expression level of miR-762,up-regulate the expression level of APC2,enhance cell radiosensitivity,inhibit cell proliferation,promote cell apoptosis,and down-regulate CyclinD1 and Bcl-2 protein expression levels in Ishikawa cells,while up-regulate the protein expression levels of p21 and Bax (P＜0.05).Furthermore,interfering with the APC2 gene significantly reversed the promoting effect of inhibiting miR-762 expression on the radiosensitivity of Ishikawa cells.Conclusion:miR-762 was highly expressed in EC cancer tissues and cell lines,and inhibiting its expression could enhance the radiosensitivity of Ishikawa cells by up-regulating the expression of APC2. |
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| Keywords: | endometrial carcinoma miR-762 adenomatosis polyposis coli 2 proliferation apoptosis radiosensitivity |
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