Application of miR-135b inhibitor to medicament for treating lung metastasis and relapse of osteosarcoma

1. The technology of mir-135b and inhibitors, applied in gene therapy, antineoplastic drugs, drug combination, etc., can solve problems such as difficulties in the treatment of osteosarcoma lung

Inactive Publication Date: 2017-05-24
许成雄 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the present invention is to solve the current problem of difficult treatment of osteosarcoma lung metastasis and recurrence, and provides a use of miR-135b inhibitors in the treatment of osteosarcoma lung metastasis and recurrence. The use is to promote osteosarcoma lung metastasis and recurrent miR-135b as therapeutic targets, using miR-135b inhibitors to treat lung metastasis and recurrence of osteosarcoma, wherein miR-135b inhibitors include miR-135b antisense or antagomiR or plasmids expressing miR-135b antisense

Method used

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  • Application of miR-135b inhibitor to medicament for treating lung metastasis and relapse of osteosarcoma
  • Application of miR-135b inhibitor to medicament for treating lung metastasis and relapse of osteosarcoma
  • Application of miR-135b inhibitor to medicament for treating lung metastasis and relapse of osteosarcoma

Examples

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Effect test

Embodiment 1

[0023] Example 1: Treatment of miR-135b antisense significantly inhibits tumor formation of osteosarcoma tumor stem cells in animals

[0024] (1) Using the antibody of the tumor stem cell marker protein CD133, the tumor stem cells with high expression of CD133 were screened from osteosarcoma cells Saos-2 by flow cytometry.

[0025] (2) The tumor stem cells with high expression of CD133 screened above were divided into two groups, one group of cells transfected with negative control nucleotide chain (control group); the other group of cells transfected with miR-135b antisense (treatment group).

[0026] (3) After 12 hours of transfection, the transfected cells were inoculated subcutaneously on the back of five 6-week-old female nude mice (1×10 5 cells / mouse). The control cells were inoculated into the right side of the back of the nude mice; the cells of the treatment group were inoculated into the left side of the back of the nude mice, and observed for 2 months. Two months af...

Embodiment 2

[0027] Example 2: miR-135b antisense significantly inhibits lung metastasis of osteosarcoma

[0028] (1) In order to establish an animal model of osteosarcoma lung metastasis, a stable osteosarcoma cell line with low expression of miR-135b was first established.

[0029] 1 × 10 in a six-well plate 6 The density of cells / well was seeded with LM5 osteosarcoma cells. On the second day after cell plating, the plasmid expressing miR-135b antisense or the control plasmid expressing non-specific nucleotide single strand was transfected into the cells. Selection of stable cells began 48 hours after transfection. The cells were treated with 2ug / ml puromycin every three days for a total of 3 times. On the twelfth day of screening, some cells were harvested, and RNA was extracted to detect the expression level of miR-135b. After confirming the low expression of miR-135b ( figure 2 A), using the stable cells of this line to establish an animal model of osteosarcoma lung metastasis. ...

Embodiment 3

[0031] Example 3: miR-135b antagomiR significantly inhibits the recurrence of osteosarcoma caused by tumor stem cells. Establishment of tumor bearing tumor model in nude mice: The tumor stem cells with high expression of CD133 were selected from Saos-2 osteosarcoma cells by flow cytometry using the antibody of tumor stem cell marker protein CD133, and the cells were inoculated into 18 6-week-old female nude mice. Subcutaneously on the back of the mouse (1×10 5 cells / mouse). Two weeks later, the nude mice were divided into three treatment groups according to the tumor size, and the drugs were treated by intraperitoneal injection. The first group was injected with phosphate buffered saline (100ul) (untreated); the second group of animals was injected with cisplatin (5mg / Kg body weight) and 10uM negative control nucleotide chain (CDDP); the treatment group of animals was injected with cisplatin and 10 uM miR-135bantagomiR (CDDP-miR-135b-in). Drug treatment was performed every ...

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Abstract

The invention provides application of a miR-135b inhibitor to a medicament for treating lung metastasis and relapse of osteosarcoma. According to the application, miR-135b is taken as a treatment target point, and the lung metastasis and relapse of the osteosarcoma are inhibited by using the miR-135b inhibitor (antisense or antagomiR or a plasmid expressing miR-135b antisense).

Description

technical field [0001] The invention relates to the application of a miR-135b inhibitor in the medicine for treating lung metastasis and recurrence of osteosarcoma. Background technique [0002] Osteosarcoma is a primary malignant bone tumor that mainly occurs in the 10-25 age group with high morbidity and mortality. In the early 1980s, the 5-year survival rate of osteosarcoma patients had reached 60%-70%, but it has not been further improved in the past 30 years. The main causes of treatment failure in osteosarcoma are metastasis and recurrence. According to reports, the survival rate of patients with osteosarcoma is only 20-30% once metastasis or recurrence occurs. Lung metastasis is the main way of osteosarcoma metastasis. Among newly diagnosed patients, 20% of patients had distant metastasis, 90% of which were lung metastases; 50% of patients receiving regular treatment had recurrence, of which 80% were pulmonary metastases, suggesting resolution of osteosarcoma lung ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K31/7105A61P35/04
CPCA61K31/7105
Inventor 许成雄金花许猛
Owner 许成雄
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