| 陈倩倩,张英丽,李阳,等.细胞周期蛋白依赖性激酶抑制剂 2B反义 RNA1靶向微小 RNA-339-5p对乳腺癌细胞增殖、迁移、侵袭的影响[J].安徽医药,2022,26(7):1390-1394. |
| 细胞周期蛋白依赖性激酶抑制剂 2B反义 RNA1靶向微小 RNA-339-5p对乳腺癌细胞增殖、迁移、侵袭的影响 |
| Effects of CDKN2B-AS1 on breast cancer cell proliferation, migration, and invasion by targeting miR-339-5p |
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| DOI：10.3969/j.issn.1009-6469.2022.07.027 |
| 中文关键词: 乳腺肿瘤 细胞周期蛋白依赖性激酶抑制剂 2B反义 RNA1 微小 RNA-339-5p（miR-339-5p） 增殖 迁移 侵袭 |
| 英文关键词: Breast neoplasms Cyclin-dependent kinase inhibitor 2B-AS1 miR-339-5p Proliferation Migration Invasion |
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| 中文摘要: |
| 目的探讨长链非编码 RNA细胞周期蛋白依赖性激酶抑制剂 2B反义 RNA1(lncRNA CDKN2B-AS1)对乳腺癌细胞增殖、迁移、侵袭的影响及分子机制。方法收集南阳市中心医院 2017年 1月 2019年 1月收治的 37例乳腺癌患者的癌组织及癌旁组织。将 MDA-MB-453细胞分为 CDKN2B-AS1阴性对照( si-NC组)、 CDKN2B-AS1小分子干扰 RNA(si-CDKN2B-AS1组)、 CDKN2B-AS1小分子干扰 RNA+miR-339-5p阴性对照( si-CDKN2B-AS1+anti-miR-NC组)以及 CDKN2B-AS1小分子干扰 RNA+ miR-339-5p特异性寡核苷酸抑制剂( si-CDKN2B-AS1+anti-miR-339-5p组)。采用 RT-qPCR法对 CDKN2B-AS1及微小 RNA-339-5p(miR-339-5p)表达水平进行检测；细胞周期及增殖活性检测分别采用流式细胞术及 MTT实验；采用 Transwell小室技术对细胞迁移和侵袭进行检测；采用 Western blotting法对增殖标记蛋白细胞增殖核抗原 -67(Ki67)、细胞周期依赖性蛋白激酶抑制因子 1A(P21)、上皮型钙黏蛋白( E-cadherin)、神经型钙黏蛋白(N-cadherin)蛋白表达进行检测；荧光素酶报告实验检测 CDKN2B-AS1对 miR-339-5p的靶向调控。结果在乳腺癌组织中 CDKN2B-AS1表达水平上调［( 2.23±0.08)比( 1.00±0.06)］(P<0.05)。抑制 CDKN2B-AS1可增加 G0期细胞比例［(43.29±3.76)%比( 30.25±3.01)%］、 P21表达水平升高， S期细胞比例［(21.91±3.10)%比( 34.19±3.32)%］、细胞存活率［( 53.02±5.38)%比( 100.00±7.12)%］、迁移、侵袭数以及 Ki67、E-cadherin、N-cadherin蛋白表达水平降低( P<0.05)。 CDKN2B-AS1靶向调控 miR-339-5p，抑制 miR-339-5p逆转抑制 CDKN2B-AS1对 MDA-MB-453细胞增殖、迁移、侵袭的作用。结论抑制 CDKN2B-AS可抑制乳腺癌 MDA-MB-453细胞增殖、迁移、侵袭，且靶向调控 miR-339-5p表达。关键词:乳腺肿瘤；细胞周期蛋白依赖性激酶抑制剂 2B反义 RNA1；微小 RNA-339-5p(miR-339-5p)；增殖；迁移；侵袭 |
| 英文摘要: |
| Objective To investigate the effect of Long non-coding RNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (ln-cRNA CDKN2B-AS1) on the proliferation, migration and invasion of breast cancer cells and its molecular mechanism.Methods The cancer tissues and paracancerous tissues of 37 breast cancer patients who were admitted to Nanyang Central Hospital from January2017 to January 2019 were collected. MDA-MB-453 cells were assigned into CDKN2B-AS1 negative control (si-NC group), CDKN2B-AS1 small interfering RNA (si-CDKN2B-AS1 group), CDKN2B-AS1 small interfering RNA+miR-339-5p Negative control (si-CDKN2B-AS1+anti-miR-NC group) and CDKN2B-AS1 small interfering RNA+miR-339-5p specific oligonucleotide inhibitor (si-CDKN2B-AS1+ anti-miR-339 -5p group). The expression levels of CDKN2B-AS1 and Mir-339-5p were detected by RT-qPCR. The cell cycle and prolif-eration activity were detected by flow cytometry and MTT assay. Cell migration and invasion were detected by Transwell chamber tech-nique. The expressions of the proliferation marker protein proliferating nuclear antigen-67 (Ki67), cyclin-dependent protein kinase in-hibitor 1A (P21), epithelial cadherin (E-cadherin) and N-cadherin proteins were detected by Western blot. The targeted regulation of Mir-339-5p by CDKN2B-AS1 was detected by luciferase reporting assay.Results The expression level of CDKN2B-AS1 was up-regu-lated in breast cancer tissues [(2.23±0.08) vs. (1.00±0.06)](P<0.05). Inhibition of CDKN2B-AS1 could increase the proportion of cells in G0 phase [(43.29±3.76)% vs. (30.25±3.01)%], increase the expression level of P21, decrease the proportion of cells in S phase [(21.91±3.10)% vs. (34.19±3.32)%], survival rate [(53.02±5.38)% vs. (100.00±7.12)%], migration and invasion number, and decrease the expression levels of Ki67, E-cadherin and N-cadherin (P<0.05). CDKN2B-AS1 could target and regulate miR-339-5p, and inhibit-ing miR-339-5p could reverse the effect of inhibiting CDKN2B-AS1 on the proliferation, migration and invasion of MDA-MB-453 cells. Conclusions Inhibition of CDKN2B-AS can inhibit the proliferation, migration and invasion of breast cancer MDA-MB-453 cells, and target the expression of miR-339-5p. |
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