Small molecular non-coding RNA gene hsa-miR-29b and use thereof
A non-coding, small-molecule technology applied in the field of RNA and its applications
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Embodiment 1
[0058] Example 1: In Vitro Tube Formation Experiment
[0059] The in vitro tube formation experiment is an experiment in which vascular endothelial cells form a lumen structure in vivo by simulating in vitro. Angiogenesis involves the degradation of vascular basement membrane, the proliferation and migration of vascular endothelial cells into the extracellular matrix and the reorganization of vascular endothelial cells into a three-dimensional tubular structure to form a new basement membrane. In the in vitro tube formation experiment, human umbilical vein endothelial cells HUVEC were provided with extracellular matrix by laying matrigel to simulate the in vivo environment. Human umbilical vein endothelial cells HUVEC formed a two-dimensional tubular structure through processes such as proliferation, migration and adhesion. Budding is an essential step in angiogenesis. Therefore, in vitro, the budding ability of HUVEC can be evaluated by calculating the number of nodes of the ...
Embodiment 2
[0100] Embodiment 2 Tumor formation and immunohistochemical experiments in nude mice
[0101] 2.1 Cell transfection
[0102] Use Invitrogen’s transfection reagent Lipofectamine RNAi MAX to transfect the target RNA (hsa-miR-29b or NC) into the hepatocellular carcinoma cell line LM6 (LM6 for short, from Shanghai Cell Institute), the final concentration of the target RNA is 50nM, after 48 hours of culture Digested with trypsin and counted according to 1 × 10 7 The concentration of cells / ml was resuspended in PBS. Reagents and materials not indicated in specific sources in this example are commercially available standard samples.
[0103] 2.2 Tumor formation in nude mice
[0104] Five 5-week-old BALB / c nude mice were selected and subcutaneously injected with 1×10 6 Restoration of hsa-miR-29b or NC expression in LM6 tumor cells. Continuously observe and measure the size of the tumor for more than 1 month. The tumor tissues of all nude mice were completely stripped, and made i...
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