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Application of pax3-fkhr and acta1 gene in prevention and treatment of rhabdomyosarcoma

A rhabdomyosarcoma and gene technology, applied in the field of biomedicine, can solve the problems that there are no exact and effective methods and means for the diagnosis, treatment and prognosis of this type of disease, and the mechanism of occurrence is not completely clear.

Active Publication Date: 2022-04-01
SHANGHAI EAST HOSPITAL EAST HOSPITAL TONGJI UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the pathogenesis of this disease is still not completely clear, and there are no exact and effective methods and means for the diagnosis, treatment and prognosis of this type of disease.

Method used

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  • Application of pax3-fkhr and acta1 gene in prevention and treatment of rhabdomyosarcoma
  • Application of pax3-fkhr and acta1 gene in prevention and treatment of rhabdomyosarcoma
  • Application of pax3-fkhr and acta1 gene in prevention and treatment of rhabdomyosarcoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1. Study on the expression of ACTA1 in different rhabdomyosarcoma cells

[0057] In this example, the inventors detected the protein expression of ACTA1 in different rhabdomyosarcoma cells. The specific experimental process is as follows: select acinar rhabdomyosarcoma cells RH30 (SJ-RH30), RH4 (SJ-RH4), H41 (SJ-RH41); embryonic rhabdomyosarcoma cells RD and mouse myoblasts C2C12, RIPA lysate extraction Total protein. The mouse skeletal muscle tissue lysate sample enriched in ACTA1 was used as a control. 10% SDS-PAGE electrophoresis, membrane transfer, Western blot analysis, and ACTA1 antibody was used to detect the distribution of ACTA1 expression in each sample.

[0058] The result is as figure 1 Shown, where m.sk is mouse skeletal muscle tissue lysate; RH4, RH30 and RH41 are alveolar rhabdomyosarcoma cells; RD is embryonal rhabdomyosarcoma; C2C12 is mouse myoblasts.

[0059] from figure 1 It can be seen that ACTA1 protein is expressed in different rhabdo...

Embodiment 2

[0060] Example 2. Study on the influence of PAX3-FKHR on the transcriptional activity of ACTA1

[0061] In this example, the inventors detected the regulatory effect of PAX3-FKHR on the transcriptional activity of ACTA1. The specific experimental process is as follows: using the polymerase chain reaction (PCR) method to amplify the ACTA1 gene promoter from the genomic DNA of RH30 cells, inserting it into the pLuc-MCS vector, and constructing the reporter gene plasmid; and the pRL-TK plasmid, transfected into RH30 cells using lipofetamine 2000 reagent, cultured in 0.3% low-concentration serum DMEM medium for 24-48hr, using Promega company dual luciferase activity detection kit to measure reporter gene activity, analysis Regulation of ACTA1 by PAX3-FKHR.

[0062] The inventors analyzed the transcriptional activity of the ACTA1 promoter in RH30 cells transfected with different doses of PAX3-FKHR expression plasmids as described above. The result is as figure 2 As shown, the i...

Embodiment 3

[0064] Example 3. Effect of PAX3-FKHR on ACTA1 protein expression

[0065] In this example, the inventors tested whether PAX3-FKHR has an inhibitory effect on ACTA1 protein expression. The specific experimental process was as follows: RH4 cells with high ACTA1 expression were selected and transfected with different doses of PAX3-FKHR gene expression plasmids. After 48 hours of low serum incubation, total protein was extracted and quantified. As described in Example 1, SDS-PAGE electrophoresis, membrane transfer, and Western blot analysis were performed, and α-Tubulin was used as an internal reference to detect and analyze the expression changes of ACTA1 protein and PAX3-FKHR protein itself.

[0066] The results are shown in image 3 . The Western blot analysis showed that in the alveolar rhabdomyosarcoma cells RH4 transfected with different amounts of PAX3-FKHR expression plasmids, the expression level of ACTA1 protein was inhibited with the increase of transfected plasmids....

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Abstract

The present invention provides inhibitors or antagonists of PAX3-FKHR gene or its encoded protein, activator or accelerator of RhoA / MKL1 / SRF signaling pathway and ACTA1 gene or its encoded protein in screening and preparing striated muscle Use in a therapeutic agent for sarcoma, preferably alveolar rhabdomyosarcoma. The present invention discloses a mechanism of the occurrence and development of alveolar rhabdomyosarcoma and the potential clinical application value of the mechanism in rhabdomyosarcoma, preferably in the treatment and prognosis of alveolar rhabdomyosarcoma.

Description

technical field [0001] The invention relates to the field of biomedicine. Specifically, the present invention relates to the use of PAX3-FKHR gene in preventing and treating rhabdomyosarcoma, preferably alveolar rhabdomyosarcoma. Background technique [0002] Rhabdomyosarcoma is a malignant tumor originating from striated muscle cells or mesenchymal cells differentiated into rhabdomyosarcoma, and is the most common type of soft tissue sarcoma in children. The incidence of rhabdomyosarcoma is second to malignant fibrous histiocytoma and liposarcoma, ranking third in soft tissue sarcomas. Rhabdomyosarcoma is divided into embryonal rhabdomyosarcoma, alveolar rhabdomyosarcoma and pleomorphic rhabdomyosarcoma. The etiology of rhabdomyosarcoma is still unclear. It is a malignant soft tissue tumor composed of rhabdomyoblastic cells with different degrees of differentiation, which may be related to genetic factors, chromosomal abnormalities, gene fusion and other factors. [0003...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574A61K45/00A61P35/00
CPCA61K45/00A61P35/00G01N33/57407G01N33/57484C12Q1/6886C12Q2600/136C12Q2600/158
Inventor 徐俊胡千德
Owner SHANGHAI EAST HOSPITAL EAST HOSPITAL TONGJI UNIV SCHOOL OF MEDICINE
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