丁保锋

【摘要】 目的：探討miR-577靶向调控HOXA1对乳腺癌MDB-MA-231细胞增殖、侵袭和迁移的影响。方法：将体外培养的MDB-MA-231细胞分为对照组（未处理）、miR-NC组（转染miR-577模拟物阴性对照）、miR-577组（转染miR-577模拟物）、miR-577+pcDNA组（转染miR-577模拟物和pcDNA3.1质粒）和miR-577+HOXA1组（转染miR-577模拟物和pcDNA3.1-HOXA1质粒），采用实时荧光定量PCR测定MDB-MA-231细胞中miR-577和HOXA1 mRNA的表达，Western blot测定HOXA1蛋白表达，荧光素酶报告基因实验测定miR-577和HOXA1的靶向关系，CCK-8法测定MDB-MA-231细胞增殖，流式细胞仪测定细胞周期分布，Transwell小室测定MDB-MA-231细胞侵袭和迁移。结果：与对照组、miR-NC组相比，miR-577组细胞中miR-577表达水平、G0/G1期细胞百分比升高，HOXA1 mRNA和蛋白表达水平、48与72 h细胞增殖活力、S期细胞百分比、侵袭细胞数和迁移细胞数均降低，差异均有统计学意义（P<0.05）。与miR-577组、miR-577+pcDNA组相比，miR-577+HOXA1组中HOXA1 mRNA和蛋白表达水平、48与72 h细胞增殖活力、S期细胞百分比、侵袭细胞数和迁移细胞数均升高，G0/G1期百分比降低，差异均有统计学意义（P<0.05）。结论：miR-577可通过靶向HOXA1抑制MDB-MA-231细胞增殖、侵袭和迁移。

【关键词】 miR-577 乳腺癌 HOXA1 增殖 侵袭 迁移

Effects of miR-577 Inhibits Proliferation， Migration and Invasion of Breast Cancer MDB-MA-231 Cells by Targeting HOXA1/DING Baofeng. //Medical Innovation of China， 2021， 18（10）： 0-018

[Abstract] Objective： To investigate the effects of miR-577 targeting HOXA1 on proliferation， invasion and migration of breast cancer MDB-MA-231 cells. Method： MDB-MA-231 cells cultured in vitro were divided into control group （untreated）， miR-NC group （transfected negative control of miR-577 mimics）， miR-577 group （transfected miR-577mimics）， miR-577+pcDNA group （transfected miR-577 mimics and pcDNA 3.1 plasmids） and miR-577+HOXA1 group （transfected miR-577 mimics and pcDNA3.1-HOXA1 plasmids）. The expression of miR-577 and HOXA1 mRNA in MDB-MA-231 cells were detected by real-time fluorescence quantitative PCR， the expression of HOXA1 protein was measured by Western blot， the targeting relationship between miR-577 and HOXA1 was checked by double luciferase reporter gene assay， the proliferation of MDB-MA-231 cells was tested by CCK-8， the cell cycle was examed by flow cytometry， the migration and invasion of MDB-MA-231 cells were detected by Transwell chamber. Result： Compared with the control group and the miR-NC group， the expression level of miR-577 and the percentage of G0/G1 phase cells in the miR-577 group were increased， the expression levels of HOXA1 mRNA and protein， the proliferation activity of cells at 48 and 72 h， the percentage of cells in the S phase， the number of invaded cells and the number of migrated cells were decreased， the differences were statistical significance （P<0.05）. Compared with the miR-577 group and the miR-577+pcDNA group， HOXA1 mRNA and protein expression levels， proliferation activity of cells at 48 and 72 h， percentage of cells in S phase， number of invading cells and number of migrating cells in the miR-577+HOXA1 group were all increased， while percentage of cells in G0/G1 phase was decreased， the differences were statistical significance （P<0.05）. Conclusion： miR-577 can inhibit proliferation， invasion and migration of MDB-MA-231 cells by targeting HOXA1.

[Key words] miR-577 Breast cancer HOXA1 Proliferation Invasion Migration

First-authors address： The Central Hospital of Jiamusi City， Jiamusi 154002， China

doi：10.3969/j.issn.1674-4985.2021.10.004

微小RNAs（microRNAs，miRNAs）是一类广泛分布于机体血液、组织和唾液中的内源性非编码RNA，常被作为肿瘤早期诊断和预后不良的重要指标，也是肿瘤基因治疗的潜在靶分子，其可通过碱基互补配对抑制翻译或降解靶mRNA参与包括乳腺癌在内的多种肿瘤的发生发展[1]。miR-577是miRNAs中的重要成员，被证实在肝癌、结肠癌和胶质母细胞瘤等肿瘤细胞增殖、侵袭和迁移过程中发挥着重要的抑制作用[2-3]。Yin等[4]证实miR-577在乳腺癌组织中低表达，与肿瘤大小、肿瘤分期和淋巴转移等有关，可通过靶向Rab25抑制癌细胞上皮间质转化和转移;但miR-577在乳腺癌中的作用及机制并不完全清晰。同源盒A1（HOXA1）是同源盒（HOX）基因家族成员，被证实在乳腺癌中异常高表达，通过调控癌细胞增殖、侵袭和迁移等促进肿瘤进展[5]。有研究指出，miR-577可通过靶向调控HOXA1表示抑制肝癌细胞侵袭和迁移，但其是否可能通过靶向调控HOXA1影响乳腺癌的发生发展尚不清楚[6]。……