Application of tumor-derived exosome in preparation of medicines for promoting osteoclast differentiation and osteolysis

Abstract

The invention belongs to the technical field of cell biology, and discloses an application of exosome in preparation of a medicine for promoting osteoclast differentiation and osteolysis. A preparation method of bone-targeted prostate cancer exosome comprises the following steps: separating and culturing prostate cancer cells, and when the cells are fused to 70%, separating and culturing the prostate cancer cells; transferring the cultured cells into a conditioned medium which is collected in advance and is not subjected to exosome serum treatment; continuously culturing for 48 hours; when the cells are fused to 90%, collecting a conditioned culture supernatant; and separating by adopting a differential centrifugation method to obtain the prostate cancer cell source exosome. The prostate cancer cell exosome is effectively separated and prepared, it is verified that the prostate cancer cell exosome can target bones and be combined with bone matrix cells, in-vitro osteoclast differentiation and in-vivo osteolytic changes are caused accordingly, pathological bone remodeling is caused, and finally growth of tumors in bones is promoted.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to the use of tumor-derived exosomes in the preparation of drugs for promoting osteoclast differentiation and osteolysis. Background technique [0002] At present, exosomes are extracellular vesicles with a diameter of 30-150 nm secreted by a variety of cells and have a lipid bilayer. Existing studies have confirmed that exosomes play an important role in the signaling process between cells and organs. [0003] Isolating exosomes with high purity, good biological activity and suitable for downstream detection purposes is the primary difficulty in exosome research. According to the MISEV2018 guideline of the International Society of Extracellular Vesicles (ISEV) in 2018, there are currently a variety of isolation methods for exosomes (ultracentrifugation, sucrose density gradient centrifugation, and polyethylene glycol precipitation, etc.), There is no unified stan...

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Problems solved by technology

According to the MISEV2018 guideline of the International Society of Extracellular Vesicles (ISEV) in 2018, there are currently a variety of isolation methods for exosomes (ultracentrifugation, sucrose density gradient centrifugation, and polyethylene glycol precipitation, etc.), There is no unified standard; however, each separation method has certain disadvantages: the ultracentrifugation method has high purity but low yield and requires special equipment, but because of its high purity, it is still the first choice for exosome function research. The preferred method; the preparatory work of sucrose density gradient centrifugation is cumbersome and time-consuming, and the number of exosomes obtained is small; although the polyethylene glycol precipitation method has a high exosome yield, it cannot meet the requirements due to the introduction of impurities during the precipitation process. Requirements for Subsequent Analysis of Exosome Contents

[0006] (1) The isolation of bone-targeted exosomes has not been reported;

[0007] (2) The role and mechanism of prostate cancer exosomes in the dialogue between prostate cancer and bone are unclear

[0008] The difficulty in solving the above problems and defects is: it is not clear which kind of cell exosomes can target bone, so as to provide the feasibility of exosome therapy for bone diseases; prostate cancer is prone to bone metastasis, but the exosomes of prostate cancer It is not clear whether the body can be distributed in the bone in the body and exert corresponding functions through a certain mechanism

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