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超声靶向微泡破坏(UTMD)联合脂质体通过靶向微小 RNA 干扰(shRNA)Metadherin(MTDH)增加抑制增殖、迁移、侵袭和上皮间质转化(EMT)的效果。

Ultrasound-Targeted Microbubble Destruction (UTMD) Combined with Liposome Increases the Effectiveness of Suppressing Proliferation, Migration, Invasion, and Epithelial- Mesenchymal Transition (EMT) via Targeting Metadherin (MTDH) by ShRNA.

机构信息

Department of Medical Imaging, Zhangqiu Maternal and Child Health Care Hospital, Jinan, Shandong, China (mainland).

Department of Orthopedics, Zhangqiu People's Hospital, Jinan, Shandong, China (mainland).

出版信息

Med Sci Monit. 2019 Apr 10;25:2640-2648. doi: 10.12659/MSM.912955.

Abstract

BACKGROUND Reports show that ultrasound-targeted microbubble destruction (UTMD) is a promising method of gene therapy, and metadherin (MTDH) is related to the development of breast cancer. Thus, we investigated the role of MTDH in breast cancer and compared the effect of suppressing MTDH by shRNA using liposome, UTMD, or the combination of these 2 methods. MATERIAL AND METHODS Graphing of survival curves of MTDH was analyzed by bioinformatics. UTMD was conducted using an ultrasonic therapeutic apparatus. Cell counting kit-8 (CCK-8) assay was used to measure cell viability. Migration and invasion rates were measured by wound healing test and Transwell invasion assay, respectively. The expression of MTDH, E-cadherin, metastasis-associated protein-1 (MTA-1), matrix metalloproteinase (MMP)-2, and MMP-9 were measured by Western blot and qPCR. RESULTS The prognosis of breast cancer can be decreased by the high expression of MTDH, and elevated expression of MTDH was discovered in MCF-7, MCF-10A, and T47D cell lines. UTMD combined with liposome is most efficient in transfecting shRNA, clearly suppressing the expression of MTDH and thereby decreasing cell viability, migration, invasion rate, and epithelial- mesenchymal transition (EMT) processes in the MCF-7 cell line. CONCLUSIONS UTMD combined with liposome could be used as a more efficient way to transfect shRNA into cells to suppress the expression of MTDH and thus lead to the downregulation of proliferation, migration, and EMT processes of the MCF-7 cell line, showing the potential for use in gene therapy.

摘要

背景

有报道称,超声靶向微泡破坏(UTMD)是基因治疗的一种很有前途的方法,而 Metadherin(MTDH)与乳腺癌的发展有关。因此,我们研究了 MTDH 在乳腺癌中的作用,并比较了通过脂质体、UTMD 或这两种方法联合抑制 shRNA 对 MTDH 的作用。

材料与方法

通过生物信息学分析 MTDH 生存曲线的绘图。使用超声治疗仪进行 UTMD。细胞计数试剂盒-8(CCK-8)测定用于测量细胞活力。通过划痕愈合试验和 Transwell 侵袭试验分别测量迁移和侵袭率。通过 Western blot 和 qPCR 测量 MTDH、上皮钙黏附蛋白(E-cadherin)、转移相关蛋白 1(MTA-1)、基质金属蛋白酶(MMP)-2 和 MMP-9 的表达。

结果

MTDH 的高表达可降低乳腺癌的预后,在 MCF-7、MCF-10A 和 T47D 细胞系中发现了升高的 MTDH 表达。UTMD 联合脂质体在转染 shRNA 方面最有效,明显抑制了 MCF-7 细胞系中 MTDH 的表达,从而降低了细胞活力、迁移、侵袭率和上皮-间充质转化(EMT)过程。

结论

UTMD 联合脂质体可作为一种更有效的方法将 shRNA 转染入细胞,抑制 MTDH 的表达,从而下调 MCF-7 细胞系的增殖、迁移和 EMT 过程,显示出在基因治疗中的应用潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f509/6474295/ea26321829d2/medscimonit-25-2640-g001.jpg

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