超声微泡介导的 miR-503-5p 下调通过促进 SALL1 表达抑制体外 CRC 进展。
Ultrasound microbubble-mediated miR-503-5p downregulation suppressed in vitro CRC progression via promoting SALL1 expression.
机构信息
Department of Colorectal Surgery, Baoji Central Hospital, China.
Department of Ultrasonic Medicine, Tongchuan People's Hospital, China.
出版信息
Tissue Cell. 2022 Jun;76:101811. doi: 10.1016/j.tice.2022.101811. Epub 2022 Apr 27.
BACKGROUND
This study compared the effect of ultrasound microbubble-mediated miR-503-5p downregulation with that of pure liposome-mediated miR-503-5p downregulation on colorectal cancer (CRC) progression and explored the downstream mechanism.
METHODS
Bioinformatics tools were utilized to predict miR-503-5p-targeted genes and CRC progression-associated genes. MiR-503-5p and sal-like 1 (SALL1) expressions in CRC cells and tissues were analyzed by qRT-PCR and/or bioinformatics tools; their correlations with overall survival and clinicopathological features of CRC patients were presented, and their interaction was validated by dual-luciferase reporter assay. CRC cells received ultrasound microbubble-mediated miR-503-5p downregulation and/or liposome-mediated miR-503-5p downregulation or SALL1 silencing. Cell phenotype changes were evaluated by flow cytometry, as well as MTT, Wound healing, Transwell and tube formation assays. E-cadherin, N-cadherin, Vimentin, B-cell lymphoma (Bcl)- 2, Cleaved caspase-3, and SALL1 expressions in cells were analyzed by Western blot.
RESULTS
Upregulated miR-503-5p in CRC tissues and cells was detected, associated with poorer cell differentiation, easier lymph node metastasis and higher TNM stages, and related to poorer prognoses of CRC patients. Ultrasound microbubble-mediated miR-503-5p downregulation relative to pure liposome-mediated miR-503-5p downregulation better decreased viability, inhibited migration, invasion and tube formation, enhanced apoptosis, upregulated SALL1, E-cadherin and Cleaved caspase-3, and downregulated miR-503-5p, N-cadherin, Vimentin and Bcl-2 in CRC cells. SALL1 was targeted by miR-503-5p, low-expressed in CRC tissues and cells and positively related to CRC patients' survival. Silencing SALL1 exerted the opposite effects, which reversed the effects of ultrasound microbubble-mediated miR-503-5p downregulation and vice versa.
CONCLUSION
Ultrasound microbubble-mediated miR-503-5p downregulation suppressed in vitro CRC progression via promoting SALL1 expression.
背景
本研究比较了超声微泡介导的 miR-503-5p 下调与单纯脂质体介导的 miR-503-5p 下调对结直肠癌（CRC）进展的影响，并探讨了下游机制。
方法
利用生物信息学工具预测 miR-503-5p 靶向基因和与 CRC 进展相关的基因。通过 qRT-PCR 和/或生物信息学工具分析 CRC 细胞和组织中的 miR-503-5p 和 SALL1 的表达；分析其与 CRC 患者总生存期和临床病理特征的相关性，并通过双荧光素酶报告基因检测验证其相互作用。超声微泡介导的 miR-503-5p 下调和/或脂质体介导的 miR-503-5p 下调或 SALL1 沉默处理 CRC 细胞。通过流式细胞术、MTT 实验、划痕愈合实验、Transwell 实验和管形成实验评估细胞表型变化。Western blot 检测细胞中 E-cadherin、N-cadherin、Vimentin、B-cell lymphoma (Bcl)-2、Cleaved caspase-3 和 SALL1 的表达。
结果
检测到 CRC 组织和细胞中上调的 miR-503-5p 与较差的细胞分化、更易发生淋巴结转移和更高的 TNM 分期相关，与 CRC 患者的预后较差相关。与单纯脂质体介导的 miR-503-5p 下调相比，超声微泡介导的 miR-503-5p 下调更能降低细胞活力，抑制迁移、侵袭和管形成，增强细胞凋亡，上调 SALL1、E-cadherin 和 Cleaved caspase-3，下调 CRC 细胞中的 miR-503-5p、N-cadherin、Vimentin 和 Bcl-2。SALL1 是 miR-503-5p 的靶基因，在 CRC 组织和细胞中低表达，并与 CRC 患者的生存呈正相关。沉默 SALL1 则产生相反的效果，逆转了超声微泡介导的 miR-503-5p 下调的效果，反之亦然。
结论
超声微泡介导的 miR-503-5p 下调通过促进 SALL1 表达抑制体外 CRC 进展。
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