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Application of miR-491 in preparing medicines for treating osteosarcoma

A mir-491, 1.mir-491 technology, applied in the field of medicine and biology, to achieve the effect of strengthening the anti-cancer effect

Inactive Publication Date: 2018-02-16
THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, its effect on lung metastasis of osteosarcoma has not been reported

Method used

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  • Application of miR-491 in preparing medicines for treating osteosarcoma
  • Application of miR-491 in preparing medicines for treating osteosarcoma
  • Application of miR-491 in preparing medicines for treating osteosarcoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Example 1: miR-491 significantly promotes the inhibitory effect of cisplatin (CDDP) on the growth of osteosarcoma cells

[0042] (1) Using transfection reagent lipofectamine 2000, artificially synthesized negative control nucleotide chain (NC) or miR-491mimic, which mimics miR-491, was transfected into osteosarcoma cells U2OS and Saos-2.

[0043] (2) 24 hours after transfection, the cells were transferred to a 96-well plate (5000 cells / well, 12 wells / group) and incubated. After incubation for 12 hours, the cells were treated with 0.1% DMSO or 10uM CDDP (cisplatin) for 48 hours, and the cell growth was detected by MTT kit.

[0044] The osteosarcoma tumor cells transfected with chemically synthesized negative control nucleotide chain and miR-491 were used to observe the sensitivity of miR-491 to osteosarcoma cell invasion and cisplatin. Experimental results such as figure 1 As shown, miR-491 significantly inhibited the invasion ability of osteosarcoma cells and enhanced...

Embodiment 2

[0045] Example 2: miR-491 significantly promotes the pro-apoptotic effect of cisplatin on osteosarcoma cells

[0046] (1) Using transfection reagent lipofectamine 2000, artificially synthesized negative control nucleotide chain (NC) or miR-491mimic was transfected into osteosarcoma cells U2OS and Saos-2.

[0047] (2) After 24 hours of transfection, the cells were transferred to a 6-well plate culture dish, and the cell density in the culture dish was about 70% (70% refers to the area of ​​the culture dish occupied by the cells after growth), and carried out Incubation. After incubation for 12 hours, the cells were treated with 0.1% DMSO or 10 uM CDDP (cisplatin) for 24 hours, and apoptotic cells were detected using an apoptosis kit.

[0048] Experimental results such as figure 1 As shown, the treatment of miR-491 significantly promoted the pro-apoptotic effect of cisplatin on osteosarcoma cells.

Embodiment 3

[0049] Example 3: miR-491 inhibits the invasion ability of osteosarcoma cells

[0050] (1) U2OS and Saos-2 osteosarcoma cells were transfected with miR-491 mimic or negative control nucleotides and cultured for 24 hours.

[0051] (2) Treat the Transwell for invasion with serum-free cell culture medium at 37 degrees for 3 hours.

[0052] (3) Add 600 ul of cell culture medium with 10% serum to the culture dish of the 24-well plate, and put the Transwell into the 24-well plate.

[0053] (4) Digest the transfected cells and suspend them in serum-free culture medium. Adjust the cell concentration to 4×10 using serum-free medium 5 cells / ml, and add 100ul to each Transwell, place at 37°C, 5% CO 2 incubated in a cell culture incubator.

[0054] (5) After 24 hours of incubation, suck off the culture medium in the Transwell, wipe off the non-penetrated cells on the upper surface of the Transwell with a wet cotton swab, and soak the Transwell with 0.1% methanol crystal violet solutio...

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Abstract

The invention relates to an application of miR-491 in preparing medicines for treating osteosarcoma. According to the application, miR-491 is used for preparing medicines for inhibiting osteosarcoma growth, pulmonary metastasis and pulmonary metastatic relapse. MiR-491 and a pharmaceutically acceptable carrier are bonded, and the bonded MiR-491 and pharmaceutically acceptable carrier are compounded with antitumor drugs and / or pharmaceutically acceptable auxiliary materials to prepare the medicinal composition. For the medicinal composition, miR-491 is adopted as the active ingredient, whereinthe nucleotide sequence of miR-491 is shown as SEQ ID No.1. The medicinal composition contains miR-491 bonded to the carrier and / or the pharmaceutically acceptable auxiliary materials. The in-vivo andin-vitro experiments prove that after overexpression of miR-491, the growth of osteosarcoma and pulmonary metastasis can be effectively inhibited, the inhibiting effect and apoptosis-promoting effectof cis-platinum for osteosarcoma cells can be promoted, which means that miR-491 enhances the anticancer effect of platinum-containing medicines, and can be used for preparing novel treatment medicines for treating osteosarcoma cell pulmonary metastasis, and thus possibility is provided for clinical treatment of pulmonary metastatic osteosarcoma.

Description

technical field [0001] The invention relates to the fields of medicine and biotechnology, in particular to the application of a microRNA in the preparation of medicine for treating osteosarcoma. Background technique [0002] Osteosarcoma (Osteosarcoma) is a malignant tumor originating from the bone. It is the most common type of bone malignancy. -25 years old age group. 75% of osteosarcoma patients are young male patients, and the prognosis is extremely poor. The 5-year survival rate of patients after surgery is 60%-70%, which has not been further improved in recent years. The main causes of treatment failure are metastasis, relapse and drug resistance. According to literature reports, 30% of osteosarcoma patients develop drug resistance to chemotherapy, and 80% of osteosarcoma patients die due to metastasis. According to reports, the survival rate of osteosarcoma patients once metastasized is only 20-30%. Lung metastasis is the main way of osteosarcoma metastasis. Amon...

Claims

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Application Information

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IPC IPC(8): A61K31/7088A61K31/7105A61K33/24A61P35/00A61P35/04
CPCA61K31/7088A61K31/7105A61K33/24A61K2300/00
Inventor 许成雄金花许猛罗松李清王东
Owner THE THIRD AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIV OF PLA
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