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Application of E2F1 protein in preparation of medicine for diagnosing and treating osteosarcoma

A technology for E2F1 and osteosarcoma, applied in gene therapy, antineoplastic drugs, drug combinations, etc., can solve the problems of no E2F1 protein research reports, and achieve accurate diagnosis and treatment

Inactive Publication Date: 2013-12-25
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In terms of tumor research, it has been reported that E2F1 protein is related to the differentiation of many tumor cells, but there is no research report on E2F1 protein in the differentiation of osteosarcoma

Method used

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  • Application of E2F1 protein in preparation of medicine for diagnosing and treating osteosarcoma
  • Application of E2F1 protein in preparation of medicine for diagnosing and treating osteosarcoma
  • Application of E2F1 protein in preparation of medicine for diagnosing and treating osteosarcoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A total of 37 clinical samples of osteosarcoma were selected, fixed with 10% formaldehyde solution for 7 days, and then paraffin-embedded tissue blocks were prepared. Paraffin sections with a thickness of 8 μm were prepared from the samples, and the expression of E2F1 protein (purchased from Proteintech Group Inc.) was detected by immunohistochemical method, and at the same time, another section was selected for H&E staining. The expression of E2F1 protein and H&E staining were observed under an upright microscope. Such as figure 1 As shown in A, the expression of E2F1 protein in the sample cancer tissue is higher than that in the non-tumor tissue in the same slice. Statistical analysis was performed on the expression of E2F1 protein in 37 clinical samples of osteosarcoma, and it was found that 14 cases were strongly positive in cancer tissues, 9 cases were positive, 9 cases were weakly positive, and 4 cases were negative ( figure 1 B).

Embodiment 2

[0022] Further analysis of the expression of E2F1 protein and bone differentiation marker protein OPN (purchased from Santa Cruz Company) in the clinical samples of osteosarcoma in the aforementioned Example 1 revealed that when E2F1 is strongly positive in osteosarcoma tissue, OPN is Negative expression ( figure 2 A); Through statistical analysis, the expression of E2F1 and OPN was negatively correlated ( figure 2 B).

Embodiment 3

[0024] In subcultured osteosarcoma cells U2OS (purchased from the Cell Bank of the Chinese Academy of Sciences), the E2F1 gene was silenced by siRNA technology (E2F1 group), and a blank control (scramble siRNA group) was set. After 24 hours of transfection, retinoic acid (purchased from Sigma-Aldrich Company) was given for 7 days, and Western blotting, real time PCR and alkaline phosphatase staining were used respectively (alkaline phosphatase detection kit was purchased from Beyontien Biotechnology Institute) to detect the differentiation ability of cells. Such as image 3 As shown, after silencing the E2F1 gene, the differentiation level of osteosarcoma U2OS was significantly increased.

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Abstract

The invention provides application of E2F1 protein in preparation of a medicine for diagnosing and treating osteosarcoma, and application of the E2F1 protein in preparation of a product for treating osteosarcoma by virtue of differentiation therapy. The E2F1 protein is characterized in that the amino acid sequence is as shown in SEQ ID NO: 1. The experiment result shows that the expression of the E2F1 protein in an osteosarcoma tissue is obviously higher than that in a para-carcinoma tissue and is closely related to the differentiated degree of osteosarcoma. Therefore, E2F1 protein can be used as specific tag protein for diagnosing and treating osteosarcoma by virtue of the differentiation therapy; the diagnosis and treatment of osteosarcoma can be performed more accurately and quickly. The E2F1 protein can be used for providing a new treatment target and a new effective medicine for treating osteosarcoma.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals and relates to the application of E2F1 protein in the preparation of drugs for diagnosing and treating osteosarcoma. Background technique [0002] Osteosarcoma is the most common malignant bone tumor in childhood or adolescence. It is highly malignant, progresses rapidly, has a high incidence of hematogenous metastasis, and has a very poor prognosis. At present, the treatment of osteosarcoma mostly adopts a multidisciplinary comprehensive treatment mode with surgery as the mainstay and chemotherapy and radiotherapy as the auxiliary treatment. Although chemotherapy plays a pivotal role in the treatment of osteosarcoma, severe toxic side effects and primary or secondary drug resistance caused by high-dose chemotherapy based on the high degree of malignancy of osteosarcoma often lead to the failure of osteosarcoma chemotherapy. With the development of related disciplines and technologies, osteosarc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K38/00A61K45/00A61P35/00G01N33/68C12Q1/68
Inventor 何俏军杨波应美丹曹戟杨晓春周倩
Owner ZHEJIANG UNIV
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