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Application of ROCK2 to preparation of TRAIL reverse tolerance medicines for treating reverse osteosarcoma cells

A technology for osteosarcoma cells and osteosarcoma, which is applied in the field of biomedical testing to improve the effect of targeted therapy

Active Publication Date: 2019-11-05
THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

Studies have confirmed that ROCK2 is highly expressed in many different tumor tissues such as liver cancer, breast cancer, prostate cancer, and lung cancer. Our previous studies have also proved that ROCK2 is highly expressed in liver cancer, kidney cancer, and colon cancer. However, ROCK2 The expression in osteosarcoma tissue, and its impact on the resistance of osteosarcoma cells to TRAIL, there is no related research and report

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  • Application of ROCK2 to preparation of TRAIL reverse tolerance medicines for treating reverse osteosarcoma cells
  • Application of ROCK2 to preparation of TRAIL reverse tolerance medicines for treating reverse osteosarcoma cells
  • Application of ROCK2 to preparation of TRAIL reverse tolerance medicines for treating reverse osteosarcoma cells

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[0041] specific implementation plan

[0042] The present invention is described in further detail below through examples, but the present invention is not limited only to the following examples.

[0043] 1. Cell culture

[0044] Human osteosarcoma cells U2-OS and MG-63 were placed in DMEM high-glucose medium containing 10% fetal bovine serum (10% by volume) in 5% CO 2 , Cultured in a 37°C cell culture incubator.

[0045] 2 Expression of ROCK2 in osteosarcoma

[0046]In order to investigate the expression of ROCK2 in osteosarcoma tissues, we first collected 62 cases of osteosarcoma tissues and 10 cases of normal bone tissues, and analyzed the ROCK2 protein in 62 cases of osteosarcoma tissues and 10 cases of normal bone tissues by western blot. Compared with 10 cases of normal bone tissue, 40 cases of high expression of ROCK2 and 22 cases of low expression of ROCK2 were found. The analysis with clinical data is shown in Table 1.1. The expression of ROCK2 in osteosarcoma tissu...

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Abstract

The invention relates to an application of ROCK2 to preparation of TRAIL reverse tolerance medicines for treating reverse osteosarcoma cells. The application comprises the steps of (1) firstly, analyzing expression of ROCK2 in osteosarcoma specimens and precancerous lesions by using fluorescent quantitative PCR, western blot and immunohistochemistry, and then analyzing the relationship of the expression and prognosis; and (2) adopting an interference technique, respectively transmitting negative control and plasmid for ROCK2 interference into osteosarcoma cells U2-OS and MG-63, adding TRAIL, then transmitting negative control and raised ROCK2 slow viruses into an osteosarcoma cell line U2-OS, and analyzing the sensibility of TRAIL in the osteosarcoma cells by western blot, immunofluorescence, CCK8, Edu and flow cytometry. Through the experiment, the activity of ROCK2 kinase is restrained after clear interference of ROCK2 to reinforce the sensibility of TRAIL, so that scientific basis is provided for revealing a new treatment target point and effectively reinforcing the sensibility of the TRAIL. The ROCK2 has the advantages that reverse osteosarcoma cells have reverse tolerance to TRAIL, the new target point is provided for osteosarcoma treatment, and scientific basis is provided for effective confrontation of the TRAIL reverse tolerance.

Description

technical field [0001] The invention relates to the technical field of biomedical detection, in particular to the technical field of research on factors affecting drug resistance of osteosarcoma cells to TRAIL, in particular to a Rho-associated coiled-coil forming protein kinase 2 used in the preparation of drugs for reversing drug resistance of osteosarcoma cells to TRAIL Applications. Background technique [0002] Osteosarcoma (OS) is a primary malignant tumor of bone characterized by spindle-shaped stromal cells that can directly form bone and produce bone matrix. Poor, the 5-year survival rate is about 65%. Because the occurrence of OS seriously threatens the health of adolescents, although the research on the mechanism of chemotherapeutic drug resistance in osteosarcoma has been carried out for many years, there is still no major breakthrough, and the long-term survival rate of chemoresistant osteosarcoma patients is still less than 20%. [0003] Clinically, patients ...

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/867G01N33/68G01N33/574
CPCC12Q1/6886C12N15/86G01N33/6863G01N33/57407C12N2740/15043C12Q2600/118C12Q2600/106G01N2510/00G01N2333/70575
Inventor 郝亮邓雪强易轩
Owner THE SECOND AFFILIATED HOSPITAL TO NANCHANG UNIV
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