Application of SETDB1 to adjusting and controlling of cholesterol ways

A cholesterol and pathway technology, applied in the field of biomedicine, can solve the problems of high degree of malignancy, high heterogeneity, easy transfer, etc., and achieve the effect of promoting expression

Active Publication Date: 2019-03-29
SOUTHWEST UNIVERSITY +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The clinical manifestations of neuroblastoma are high degree of malignancy, high heterogeneity, easy transfer, easy recurrence, and poor prognosis. Therefore, its treatment is relatively difficult, and new approaches and methods are urgently needed to treat it.

Method used

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  • Application of SETDB1 to adjusting and controlling of cholesterol ways
  • Application of SETDB1 to adjusting and controlling of cholesterol ways
  • Application of SETDB1 to adjusting and controlling of cholesterol ways

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1. Down-regulation of SETDB1 inhibits Cholesterol pathway gene expression and Cholesterol content in neuroblastoma

[0055] Neuroblastoma cells BE2-C were cultured in DMEM / F12 medium containing 10% fetal bovine serum (FBS), and SHEP1 cells were cultured in DMEM medium containing 10% fetal bovine serum (FBS), KCNR was cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS). All cells were added with 1% penicillin and streptomycin, and subcultured when the cell growth density reached 80-90%.

[0056] Treatment of activated carbon-filtered serum: culture the cells under normal medium conditions, and replace the medium when the cell density is about 70%, replace the normal fetal bovine serum with activated carbon-filtered serum, and use SETDB1 shRNA carrier transformation 8h, 12h, Samples were taken at 24 hours, and the expression of ACAT2, HMGCR, HMGCS1, SREBF2, and SREBF1 genes related to the Cholesterol pathway was detected, and the results were as ...

Embodiment 2

[0058] Example 2, the influence of SETDB1 on the Cholesterol pathway of neuroblastoma

[0059] In order to further study the influence of SETDB1 on the Cholesterol pathway, the pLP1, pLP2, pLP / VSVG (Invitrogene) three-plasmid packaging system was used for lentiviral packaging. The lentiviral vector pCDH-CMV-MCS-EF1-Puro- SETDB1 and infection of BE2-C and SHEP1 cells, overexpression of SETDB1 in neuroblastoma, the results are as follows figure 2 shown. The results showed that overexpression of SETDB1 did not significantly increase the proliferation of neuroblastoma ( figure 2 , A), but can significantly increase the colony formation ability of soft agar ( figure 2 , B). By detecting genes related to the Cholestero pathway, it was found that upregulation of SETDB1 could significantly increase the expression of ACAT2, HMGCR, HMGCS1, SREBF2, and SREBF1 genes ( figure 2 , C).

Embodiment 3

[0060] Example 3, SETDB1 responds to Cholesterol pathway in neuroblastoma after Charcoal Stripped FBS treatment

[0061] Early studies have shown that when cells are cultured with FBS treated with Charcoal, due to the lack of lipids such as sterols, the fatty acid metabolism pathway in the cells is significantly increased, and the corresponding cholesterol pathway is also activated. In this study, the cells were cultured with FBS treated with Charcoal, and samples were taken at 8h, 24h, and 48h. The results are as follows: image 3 shown. The results showed that sterol pathway-related genes were found to be significantly upregulated in both BE2-C and KCNR cells ( image 3 , A). At the same time, the expression of SETDB1 was also significantly increased during this process, indicating that SETDB1 can be activated by Cholesterol-deficient medium ( image 3 , B), implying that SETDB1 has an important relationship with Cholesterol.

[0062] In order to clarify the role of SETD...

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Abstract

The invention discloses an application of SETDB1 to adjusting and controlling of cholesterol ways. When an SETDB1 expression is adjusted downwards, the cholesterol content can be reduced, and the expression of genes related to cholesterol ways can be adjusted downwards; when SETDB1 is in overexpression, the neuroblastoma soft agar cloning formation capacity can be improved, and the expression of genes related to the cholesterol ways can be neutralized; and SREBF2 is an activating transcription factor of the Cholesterol ways, so that when the SREBF2 is in overexpression, the expression of genesrelated to the Cholesterol ways can be notably promoted. The ways of adjusting and controlling cholesterol metabolism by the SETDB1 can be indicated to be used as a target point for diagnosis and treatment of cholesterol relevant diseases.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of SETDB1 in regulating cholesterol pathways. Background technique [0002] Neuroblastoma is one of the common extracranial solid tumors in children, accounting for about 6-10% of childhood tumors, with an annual incidence of about 1 / 100,000, which is only lower than leukemia and central nervous system tumors, and accounts for 1% of childhood cancer deaths. 15%. Neuroblastoma is clinically characterized by high malignancy, high heterogeneity, easy metastasis, easy recurrence, and poor prognosis. Therefore, its treatment is relatively difficult, and new approaches and methods are urgently needed for treatment. [0003] Tumor cell epigenetics is one of the current research hotspots, and many experimental evidences have confirmed that histone methylation has a very important impact on the occurrence and development of tumors. Epigenetics often leads to the silencing of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61P3/06
CPCA61K45/00A61P3/06C12Q1/6886C12Q2600/158
Inventor 崔红娟祝顺琴丁寒飞何荫良
Owner SOUTHWEST UNIVERSITY
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