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肿瘤抑制性微小RNA-452通过直接靶向RAB11A抑制乳腺癌细胞的迁移和侵袭。

Tumor-suppressive microRNA-452 inhibits migration and invasion of breast cancer cells by directly targeting RAB11A.

作者信息

Li Wanjun, Li Guoyin, Fan Zhigang, Liu Tao

机构信息

Department of Pathology, Affiliated 3201 Hospital of Xi'an Jiaotong University, Hanzhong, Shaanxi 723000, P.R. China.

Department of Medical Oncology, Affiliated 3201 Hospital of Xi'an Jiaotong University, Hanzhong, Shaanxi 723000, P.R. China.

出版信息

Oncol Lett. 2017 Aug;14(2):2559-2565. doi: 10.3892/ol.2017.6426. Epub 2017 Jun 20.

Abstract

Breast cancer is the most common type of malignant tumor in females, and metastasis is the most common cause of breast cancer-associated mortality. Previous studies have identified that abnormal expression of microRNAs is commonly observed in human cancer and may be crucial for cancer metastasis. In the present study, microRNA-452 (miR-452) was investigated for its ability to act as a tumor suppressor in breast cancer. miR-452 expression was quantified in breast cancer tissue samples and cell lines with reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Transwell migration and invasion assays were used to investigate the effect of miR-452 on the migration and invasion capabilities of breast cancer cells. Potential target genes of miR-452 were identified with miRanda and TargetScan. A luciferase reporter assay was performed to validate RAB11A as a putative target of miR-452, and was corroborated by RT-qPCR and western blot analyses. Finally, small interfering RNA (siRNA) was used to knockdown RAB11A expression and confirm whether miR-452 inhibited breast cancer cell migration and invasion via the negative regulation of RAB11A. The results revealed that miR-452 was downregulated in breast cancer tissues and cell lines, and that its downregulation may be associated with breast cancer metastasis, as miR-452 expression inhibited the migration and invasion capacities of breast cancer cells. RT-qPCR and western blot analyses indicated that miR-452 negatively regulated the expression of RAB11A mRNA and protein. The luciferase reporter assay revealed that miR-452 specifically bound to the 3'-untranslated region of RAB11A. Furthermore, inhibition of RAB11A with siRNA inhibited breast cancer cell migration and invasion. In conclusion, the present study has demonstrated that miR-452 may act as a tumor suppressor gene via inhibition of cell migration and invasion by targeting RAB11A in breast cancer.

摘要

乳腺癌是女性中最常见的恶性肿瘤类型,而转移是乳腺癌相关死亡的最常见原因。先前的研究已确定,微小RNA的异常表达在人类癌症中普遍存在,并且可能对癌症转移至关重要。在本研究中,对微小RNA-452(miR-452)作为乳腺癌肿瘤抑制因子的作用能力进行了研究。采用逆转录-定量聚合酶链反应(RT-qPCR)对乳腺癌组织样本和细胞系中的miR-452表达进行定量。使用Transwell迁移和侵袭试验来研究miR-452对乳腺癌细胞迁移和侵袭能力的影响。通过miRanda和TargetScan鉴定miR-452的潜在靶基因。进行荧光素酶报告基因试验以验证RAB11A是miR-452的假定靶标,并通过RT-qPCR和蛋白质印迹分析得到证实。最后,使用小干扰RNA(siRNA)敲低RAB11A表达,并确认miR-452是否通过对RAB11A的负调控来抑制乳腺癌细胞的迁移和侵袭。结果显示,miR-452在乳腺癌组织和细胞系中表达下调,并且其下调可能与乳腺癌转移有关,因为miR-452表达抑制了乳腺癌细胞的迁移和侵袭能力。RT-qPCR和蛋白质印迹分析表明,miR-452负调控RAB11A mRNA和蛋白质的表达。荧光素酶报告基因试验显示,miR-452特异性结合RAB11A的3'-非翻译区。此外,用siRNA抑制RAB11A可抑制乳腺癌细胞的迁移和侵袭。总之,本研究表明,miR-452可能通过靶向RAB11A抑制细胞迁移和侵袭而作为一种肿瘤抑制基因在乳腺癌中发挥作用。

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