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Application of a kind of mir-100 inhibitor in preparation of medicine for treating breast cancer

An inhibitor and breast cancer technology, applied in gene therapy, drug combination, antineoplastic drugs, etc., can solve problems such as physical damage, unresectable cancer, high risk of cancer recurrence, etc., and achieve the effect of increasing sensitivity

Active Publication Date: 2016-04-20
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the main means of treating breast cancer are radiotherapy, chemotherapy and surgical resection, and both radiotherapy and chemotherapy will cause great damage to the body, which is a great hidden danger to the follow-up survival of patients. Surgery is very important for patients with early breast cancer. The treatment is more effective. The middle and advanced cancer cells have spread and cannot be surgically removed. Moreover, many clinical cases have shown that surgical resection of cancer has a relatively high risk of recurrence. It is imminent to develop new treatments that are different from traditional methods. For The treatment of breast cancer and even all cancers is of vital value and significance

Method used

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  • Application of a kind of mir-100 inhibitor in preparation of medicine for treating breast cancer
  • Application of a kind of mir-100 inhibitor in preparation of medicine for treating breast cancer
  • Application of a kind of mir-100 inhibitor in preparation of medicine for treating breast cancer

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1: synthetic miR-100 antisense nucleic acid chain and negative control

[0030] The miR-100 antisense nucleic acid chain is named miR-100 inhibitor (ie AMO-miR-100), its sequence is: 5'-TTCGGATCTACGGGTT-3', and its base modification method is: (1): bold letters bases (3rd, 6th, 9th) LNA modification, (2) bases in italic letters 2'OME modification (12th, 15th), (3) all bases sulfo modification . (refer to patent CN102311956A).

[0031] The negative control (Scrambledcontrol, NC) is a sequence in which the base sequence is randomly scrambled. The GC content and base modification method of the base are consistent with the miR-100 inhibitor, and the sequence is 5'-GTCGGTTCTGATGTCA-3'.

[0032] The above sequences were all synthesized by Exiqon Company in Denmark.

Embodiment 2

[0034] (1) Human breast cancer cells SK-BR-3 (Shanghai Cell Bank, Chinese Academy of Sciences) were taken and cultured in 1640 medium containing 10% FBS to a cell density of about 40%.

[0035] (2) Use according to the instructions RNAiMAX transfection reagent, transfect miR-100 inhibitor and negative control, the final concentration is 50nM.

[0036](3) After 48 hours of transfection, the cells were examined under the microscope to observe the changes in cell morphology, and then the transfected cells were collected, and the inhibition efficiency of miR-100 inhibitor was detected by qPCR, and the AnnexinV kit (purchased from BD (Becton, USA) , Dickinson and Company) to detect the apoptosis of transfected cells, and the untransfected human breast cancer cell SK-BR-3 was used as a blank control.

[0037] see results figure 1 , figure 2 and image 3 . The results showed that miR-100 inhibitor (AMO-miR-100) could effectively inhibit the expression of miR-100 in breast canc...

Embodiment 3

[0039] (1) Human breast cancer cells SK-BR-3 were cultured in 1640 medium containing 10% FBS to a cell density of about 40%.

[0040] (2) Use according to the instructions RNAiMAX transfection reagent, transfected with different concentrations of miR-100 inhibitor (AMO-miR-100) and negative control.

[0041] (3) After 48 hours of transfection, the transfected cells were collected and treated with MTS[3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H -tetrazolium, innersalt] assay kit (purchased from Promega, USA) to detect cell viability and proliferation activity.

[0042] see results Figure 4 . The results show that miR-100 inhibitors can effectively inhibit the viability and proliferation of breast cancer cells, and with the increase of inhibitor concentration, the inhibition of cell viability and proliferation is more obvious, indicating that miR-100 inhibitors can inhibit breast The viability and proliferative activity of cancer cells, the...

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Abstract

The invention discloses application of a miR-100 inhibitor in preparation of a medicine for treating a breast cancer. The miR-100 inhibitor is a miR-100 antisense nucleic acid chain, and the sequence is 5'-TTCGGATCTACGGGTT-3'. The miR-100 inhibitor disclosed by the invention can induce apoptosis of tumor cells, growth of tumors in tumor transplanted mice can be effectively induced, the sensitivity of breast cancer cells on chemotherapy medicines is increased, the effect of chemotherapy is improved, and the miR-100 is a latent new target for the medicine for treating the breast cancer, and is a new path for medical therapy.

Description

(1) Technical field [0001] The present invention relates to an inhibitor for treating breast cancer, in particular to the application of a miR-100 inhibitor in the preparation of drugs for treating breast cancer. (2) Background technology [0002] Tumors are essentially malignant proliferations of cells with high metastatic capacity. Exogenous factors that lead to tumorigenesis include chemical, physical, tumorigenic viruses, and mycocins, etc. Endogenous factors include the body's immune status, genetic factors, hormone levels, DNA damage, activation of proto-oncogenes, and tumor suppressors. Gene inactivation, etc. In this process, the inhibition of apoptosis plays a key role. Apoptosis is a kind of programmed cell death. When a cell is stimulated by the outside world or itself, when it is damaged and cannot be repaired, it will undergo apoptosis to avoid damage. The accumulation of cells leads to the occurrence of diseases. Cancer is essentially a genetic disease. Cells...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61P35/00
Inventor 章晓波龚燚杨璐
Owner ZHEJIANG UNIV
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