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Application of long-chain non-coding RNA CRCAL-3 inhibitor in preparation of product for treating radiotherapy-resistant breast cancer

A long-chain non-coding and inhibitor technology, applied in the field of biomedicine, can solve problems such as unclear mechanism of action, lack of treatment methods, lack of research on radiotherapy-resistant breast cancer, etc., to improve radiotherapy sensitivity, improve radiotherapy effect, and solve Poor results from treatment

Active Publication Date: 2022-08-02
SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the relative lack of research on radioresistant breast cancer, its specific mechanism of action is not clear
This has led to a lack of proven treatments for breast cancer patients who develop radiation resistance

Method used

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  • Application of long-chain non-coding RNA CRCAL-3 inhibitor in preparation of product for treating radiotherapy-resistant breast cancer
  • Application of long-chain non-coding RNA CRCAL-3 inhibitor in preparation of product for treating radiotherapy-resistant breast cancer
  • Application of long-chain non-coding RNA CRCAL-3 inhibitor in preparation of product for treating radiotherapy-resistant breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Acquisition and analysis of long non-coding RNA CRCAL-3

[0047] 1. Construction of breast cancer radiotherapy-resistant cell lines

[0048] In order to solve the clinical problem of radiotherapy resistance in breast cancer, the inventors repeatedly irradiated breast cancer cell lines in a dose-escalation manner until the cells had a stable radiotherapy-resistant phenotype. Specifically, a linear accelerator is used for irradiation, 2 Gy / time, about 2-3 times a week. Further, the radiotherapy-resistant phenotype of the constructed breast cancer radiotherapy-resistant cell lines was verified, that is, the radiotherapy-resistant cell lines and their corresponding parental cell lines were respectively given doses of 0Gy, 2Gy, 4Gy, 6Gy, and 8Gy for radiotherapy. Final colony counts were done according to plate colony formation experiments and cell survival fractions were calculated. Further, the inventors also adopted the single-click multi-target model to fit the dose-su...

Embodiment 2

[0057] Design of specific LNA to knock down CRCAL-3 and validation of its efficiency

[0058] One, if image 3 As shown, CRCAL-3 is located in the nucleus, and it is difficult for ordinary siRNA to enter the nucleus to knock down CRCAL-3;

[0059] 2. Using LNA technology to design and synthesize LNA-CRCAL-3 that knocks down CRCAL-3

[0060] The LNA sequence is: 5'-T*A*C*T*G*G*T*T*G*C*A*G*G*A*T*T-3'(LNA GapmeR); such as Figure 4 As shown, the efficiency of LNA-CRCAL-3 knocking down CRCAL-3 reaches 80-90%; the common siRNA is only 50-55%, and LNA-CRCAL-3 has a more efficient and stable knockdown effect.

Embodiment 3

[0062] cell experiment

[0063] 1. Assess the effect of LNA-CRCAL-3 on the proliferation of breast cancer cells

[0064] The effect of LNA-CRCAL-3 on the proliferation ability of breast cancer cells was evaluated by MTS cell proliferation assay and plate clone formation assay. The experimental results showed that LNA-CRCAL-3 could attenuate the proliferation and clone formation ability of breast cancer cells. like Figure 5 Shown, A and B are the reduced proliferation ability of breast cancer cells after knockdown of CRCAL-3 in MDA-MB-231-Resistant and MCF-7-Resistant (detected by MTS method); C and D are in MDA-MB After knockdown of CRCAL-3 in -231-Resistant and MCF-7-Resistant, the clonogenic ability of breast cancer cells was weakened. (**p<0.01).

[0065] 2. Assessing the effect of LNA-CRCAL-3 on the radioresistance of breast cancer cells

[0066] (1) Label five six-well plates with 0Gy, 2Gy, 4Gy, 6Gy, and 8Gy respectively; collect breast cancer cells transfected with ...

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Abstract

The invention relates to the technical field of biological medicines, in particular to application of a long-chain non-coding RNA CRCAL-3 inhibitor in preparation of a product for treating radiotherapy-resistant breast cancer. The CRCAL-3 inhibitor is used for inhibiting proliferation of radiotherapy-resistant breast cancer cells, and it is found that the CRCAL-3 inhibitor can reverse radiotherapy resistance of the radiotherapy-resistant breast cancer, so that the radiotherapy-resistant breast cancer can recover sensitivity to radiotherapy. A new strategy is provided for treatment and prevention of radiotherapy resistant breast cancer, the survival rate of a patient is increased, and the survival time of the patient is prolonged. Meanwhile, on the basis of CRCAL-3 mechanism research, various treatment strategies of IGF2BP2 inhibition and UBR5 inhibition are provided, so that more effective treatment modes and drugs for resisting breast cancer by radiotherapy are provided, and development of the direction of tumor treatment by radiotherapy is promoted.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, and more particularly, to the use of a long-chain non-coding RNA CRCAL-3 inhibitor in the preparation of a product for treating radiotherapy-resistant breast cancer. Background technique [0002] Breast cancer is the most common malignant tumor in women. The 2021 Cancer Statistics Report shows that the incidence of breast cancer ranks first among all cancers. Radiation therapy is an integral part of the current comprehensive treatment of breast cancer. Radiation therapy for patients with early breast cancer after breast conserving surgery has now become the standard treatment mode for early breast cancer and has been widely used in clinical practice. This treatment method can protect the organ function of patients with breast cancer to the greatest extent; For patients undergoing total mastectomy, as long as there is more than one axillary lymph node metastasis or the primary tumor ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K45/00A61P35/00
CPCA61K45/00A61P35/00
Inventor 游凯云姚和瑞胡海罗曼莉毕卓菲毛璐会
Owner SUN YAT SEN MEMORIAL HOSPITAL SUN YAT SEN UNIV
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