CERS2 (ceramide synthase 2) gene and its expression product serving as treatment target of osteoporosis

A technology for osteoporosis and gene expression, applied in the biological field, can solve the problems that osteoporosis patients cannot be screened

Active Publication Date: 2015-12-30
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The objects of auxiliary examination are generally patients with advanced osteop

Method used

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  • CERS2 (ceramide synthase 2) gene and its expression product serving as treatment target of osteoporosis
  • CERS2 (ceramide synthase 2) gene and its expression product serving as treatment target of osteoporosis
  • CERS2 (ceramide synthase 2) gene and its expression product serving as treatment target of osteoporosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Differential expression of embodiment 1 CERS2 gene

[0064] 1. Research object:

[0065] Randomly select patients who are about to undergo marrow joint replacement, and measure their bone density. 50 patients with osteoporosis and 50 patients with normal bone density (traumatic fractures, without osteoporosis) were selected, aged 52-68 years. Questionnaires were used to investigate the subjects' lifestyle and health status.

[0066] Inclusion criteria for patients with osteoporosis: (1) Those who meet the diagnostic criteria for osteoporosis, refer to the "Recommended Diagnostic Criteria for Osteoporosis in China (Second Draft); (2) All patients have given informed consent.

[0067] Exclusion criteria for patients with osteoporosis: those with secondary osteoporosis.

[0068] 2. Obtaining bone tissue RNA

[0069] Take 1g of hard bone from the human femoral head removed by hip arthroplasty, and quickly store it in liquid nitrogen. In the laboratory, the Trizol one-st...

Embodiment 2

[0078] Differential expression of embodiment 2CERS2 protein

[0079] 1. The research object is the same as in Example 1.

[0080] 1. Preparation and cultivation of human osteoblasts

[0081] The cancellous bone was taken from the human femoral head removed by the arthroplasty, the soft tissue was removed, the bone was washed repeatedly with normal saline, after the washing solution was clarified, it was rinsed and shaken 3 times with PBS solution, and cut into 1mm 3 Fragments of the osteoblasts were isolated and purified by enzymatic digestion. Inoculate it in a 30ml culture flask (add appropriate amount of DMEM-F12 (1:1) medium and 10% fetal calf serum to the culture flask), place it at 37°C, 5% CO 2 cultured in a constant temperature incubator. After 2 days, the medium was changed and unadhered cells were removed, and the medium was changed every 3 days thereafter, and observed under an inverted microscope. After the primary cells fused into a monolayer, they were digest...

Embodiment 3

[0090] Embodiment 3 inhibits CERS2 gene expression

[0091] 1. siRNA design and synthesis

[0092] siRNA sequences targeting CERS2:

[0093] siRNA1-CERS2:

[0094] The sense strand is 5'-AACUUUCUUCAUGUCAUAGAA-3' (SEQ ID NO.7);

[0095] The antisense strand is 5'-CUAUGACAUGAAGAAAGUUUG-3' (SEQ ID NO.8),

[0096] siRNA2-CERS2:

[0097] The sense strand is 5'-UCAUGUAGUACCAAUACUGGG-3' (SEQ ID NO.9);

[0098] The antisense strand is 5'-CAGUAUUGGUACUACAUGAUU-3' (SEQ ID NO.10),

[0099] siRNA3-CERS2:

[0100] The sense strand is 5'-AAUCCUUUCGCUUGACAUCAG-3' (SEQ ID NO.11);

[0101] The antisense strand is 5'-GAUGUCAAGCGAAAGGAUUUC-3' (SEQ ID NO.12)

[0102] Negative control siRNA sequence (siRNA-NC):

[0103] The sense strand is 5'-CGUACGCGGAAUACUUCGA-3' (SEQ ID NO.13);

[0104] The antisense strand is 5'-UCGAAGUAUUCCGCGUACG-3' (SEQ ID NO.14).

[0105] Carry out in vitro culture of osteoporosis osteoblasts according to the method of embodiment 2, get the osteoblasts in logari...

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Abstract

The invention discloses a CERS2 (ceramide synthase 2) gene and its expression product serving as a molecular marker for treating osteoporosis. Judgment may be given to whether a patient with osteoporosis or a treatment subject has the risk of suffering from osteoporosis, by detecting CERS2 gene and its expression product contents of bone tissues. The study on proliferation of osteoblasts cultured in vitro discovers that the expression inhibition of the CERS2 gene may promote the proliferation of the osteoblasts and that the proliferation of the osteoblasts may also be promoted by inhibiting the function of CERS2 proteins, and results of the study show that the CERS2 gene and its expression product are potential drug targets for treating osteoporosis.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of CERS2 gene in the diagnosis and treatment of osteoporosis. Background technique [0002] Osteoporosis (osteoporosis) is a systemic bone disease, characterized by decreased bone mass and destruction of bone microstructure, manifested as increased bone fragility, thus greatly increasing the risk of fracture, even with minor trauma or Fractures are also prone to occur in the absence of trauma. Vertebral compression fractures often occur unknowingly, and can also be induced by coughing, sneezing, and minor trauma. Osteoporosis is a multifactorial chronic disease. Before the fracture occurs, there are usually no special clinical manifestations. The disease is more common in women than in men, but it can occur in all ages, and it is common in postmenopausal women and the elderly. With the increase of the elderly population in my country, the incidence of osteoporosis...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68A61K45/00A61P19/10
CPCA61K45/00C12Q1/6883C12Q2600/158G01N33/6893G01N2800/108
Inventor 李曙光杨承刚
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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