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Application of miR-125b and chemotherapeutic agent in preparation of drug for treating thyroid cancer

1. The technology of mir-125b and thyroid cancer, which is applied in the field of biomedicine, can solve problems such as tumor occurrence, and achieve the effect of enhancing sensitivity and increasing autophagy

Active Publication Date: 2018-11-02
GUANGDONG PHARMA UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Overexpression of some miRNAs may suppress tumor suppressor gene expression, while downregulation of other miRNAs may lead to tumorigenesis [6,7]

Method used

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  • Application of miR-125b and chemotherapeutic agent in preparation of drug for treating thyroid cancer
  • Application of miR-125b and chemotherapeutic agent in preparation of drug for treating thyroid cancer
  • Application of miR-125b and chemotherapeutic agent in preparation of drug for treating thyroid cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 MiR-125b is down-regulated in thyroid tumor tissue and thyroid cancer cells.

[0067] RNA from 30 pairs of thyroid tissues, including 12 follicular carcinomas and 18 follicular adenomas, was used to detect miR-125b expression levels by qRT-PCR. For details, see the "Materials and methods" section.

[0068] In the 4 pairs of follicular adenomas, the expression of miR-125b was not significantly different between tumor and non-tumor tissues. In 2 follicular carcinomas, the expression level of miR-125b was upregulated in tumor tissues compared with non-tumor tissues, while miR-125b was upregulated in the paracancerous tissues paired with tumor tissues in 10 follicular carcinomas and 14 follicular adenomas. Significantly down-regulated in tumor tissue compared to figure 1 b). We also evaluated the expression of miR-125b in human follicular thyroid carcinoma (WRO) and anaplastic thyroid carcinoma (FRO, KAT19)) and normal thyroid cells (Nthy-ori 3-1, Nthy1) cells ...

Embodiment 2

[0069] Example 2 Overexpression of miR-125b sensitizes thyroid cancer cells to cisplatin and sorafenib.

[0070] To further elucidate the role of miR-125b in thyroid cancer, we stably overexpressed miR-125b by transfecting miR-125b mimics and miR-125b inhibitors in thyroid cancer cells, and then screened the cells with puromycin . Additionally, we established a mock control (pre-con) and an inhibitor control (anti-con), and used the untransfected group (con) as a control. For details, see the "Materials and methods" section.

[0071] Overexpression / silencing of miR-125b in thyroid cancer cells confirmed by RT-PCR ( figure 2 a). These stable cells were seeded in 96-well plates, and the growth curves of the cells after administration of different concentrations of cisplatin or sorafenib for 48 hours were measured by MTT assay. Compared with the negative control, overexpressed miR-125b significantly sensitized thyroid cancer cells to cisplatin and sorafenib ( figure 2 b an...

Embodiment 3

[0072] Example 3 MiR-125b targets the 3'UTR of Foxp3 to inhibit its expression.

[0073] Bioinformatic analysis predicted Foxp3 to be a target of miR-125b. For details, see the "Materials and methods" section.

[0074] The sequence 508-526 of the 3'-UTR perfectly matched with miR-125b ( image 3 a). Targets were experimentally validated with a luciferase reporter system. like image 3 In b, co-transfected miR-125b inhibits reporter luciferase activity containing the 3′UTR sequence of wild-type Foxp3, but not mutant Foxp3, as shown by a dual-luciferase reporter assay. Further analysis revealed that miR-125b represses endogenous Foxp3 mRNA ( image 3 c) and protein levels ( image 3 d). Taken together, these data suggest that miR-125b can directly target the 3′-UTR sequence of Foxp3.

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Abstract

The invention belongs to the field of biomedicines and relates to an application of miR-125b or a mimic or an accelerant thereof in preparation of a chemotherapeutic drug synergist. Studies find thatthe expression of Foxp3 is up-regulated when miR-125b is down-regulated in a thyroid cancer tissue sample. Besides, miR-125b can be combined with 3'UTR of miR-125b to directly act on Foxp3 and inhibitexpression of Foxp3, and therefore, the negative correlation between miR-125b and Foxp3 is disclosed. Overexpressed miR-125b remarkably increases the sensitivity of sensitized thyroid cancer cells totreatment with a chemotherapeutic agent through induction, and in-vitro and in-vivo autophagy is performed through an Atg7 way. In general, miR-125b negatively controls autophagy of an Foxp3 promotednew mechanism and enhances the curative efficacy of the chemotherapeutic agent for thyroid cancer, and it indicates that miR-125b has a remarkable treatment significance in chemotherapy of the thyroid cancer.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of miR-125b or a miR-125b promoter and a chemotherapeutic agent in the preparation of antitumor drugs. Background technique [0002] Thyroid cancer accounts for 90% of endocrine malignancies and about 1% of all human malignancies. It is estimated that there were approximately 62,450 new cases of thyroid cancer in the United States in 2016. Although surgery has been the mainstay of treatment for thyroid cancer for decades, some patients still experience recurrence [1,2]. It is crucial to find potential markers and prognostic indicators for patients with thyroid cancer [3]. [0003] MicroRNA (miRNA) is an endogenous single-stranded non-coding RNA that plays an important role in various biological processes such as cells, including proliferation, adhesion and differentiation. It is estimated that more than one-third of human genes are targeted by miRNAs [4]. Increasing ev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61K33/24A61K31/44A61K31/282A61P35/00A61P13/08
CPCA61K31/44A61K31/555A61K31/7105A61K33/24A61P13/08A61P35/00A61K2300/00
Inventor 王珊珊
Owner GUANGDONG PHARMA UNIV
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