A kind of microrna and application thereof associated with tumor cell resistance to Sorafenib

A technology for fenib drug resistance and drug resistance, applied in the fields of biotechnology and medicine, can solve problems such as unclear microRNA personalized treatment, achieve the effect of improving efficiency, reducing IGF1R, and reversing tumor drug resistance

Active Publication Date: 2021-08-17
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, especially in the study of liver cancer, there is no systematic analysis of which of the numerous microRNAs in humans are the key factors regulating drug resistance, and it is not clear how to use microRNAs for personalized treatment

Method used

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  • A kind of microrna and application thereof associated with tumor cell resistance to Sorafenib
  • A kind of microrna and application thereof associated with tumor cell resistance to Sorafenib
  • A kind of microrna and application thereof associated with tumor cell resistance to Sorafenib

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: Differential expression of miR-378a-3p in Sorafenib sensitive and resistant cells

[0045] 1. Screening of differentially expressed microRNAs in Huh7 wild cells and Huh7 sorafenib-resistant cells

[0046] (1) Cell culture

[0047] The Huh7 wild cell line was cultured in DMEM medium containing 10% fetal bovine serum, and the Huh7 sorafenib-resistant cell line was cultured in a medium with a final concentration of 6 μmol / L of sorafenib. All cells were cultured in 5% CO 2 , 37 ℃ cell culture incubator.

[0048] Commissioned Shanghai Kangcheng Bioengineering Co., Ltd. to independently build microRNA libraries for Huh7 wild cells and Huh7 sorafenib-resistant cell lines (each cell provides three independent biological replicates), followed by next-generation sequencing. DESeq2 based on negative binomial distribution was used to analyze the expression of microRNAs between samples, and evaluated from two levels of difference fold (Foldchange) and p-value. The scr...

Embodiment 2

[0068] Example 2: Effect of interfering with miR-378a-3p on cell drug resistance

[0069] 1. Huh7 and HCC-LM3 drug-resistant cell lines transfected with miR-378a-3p mimic and verification

[0070] (1.1) Huh7 and HCC-LM3 drug-resistant cell lines transfected with miR-378a-3p mimic

[0071] Huh7 and HCC-LM3 drug-resistant cells in the logarithmic growth phase were digested with trypsin for 5 minutes, and then resuspended into a single cell suspension with medium. After cell counting, 2*10 5 The density of the cells was inoculated into 6-well plates, shaken evenly by cross method, and cultured in an incubator to make them adhere to the wall. Add the pre-mixed miR-378a-3p mimic and ctrl transfection reagents purchased from Guangzhou Ruibo Biotechnology Co., Ltd. (see Table 6) after attachment, and replace the DMEM medium with 10% fetal bovine serum after 8 hours . After 16 hours, Huh7 and HCC-LM3 drug-resistant cells transfected with miR-378a-3p mimic were obtained. The same a...

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Abstract

The invention discloses a microRNA related to sorafenib drug resistance in hepatocellular carcinoma and its application. The invention selects Huh7 and HCC‑LM3 wild and their Sorafenib drug resistant cell lines as research objects. Sequencing, verification and optimization, to obtain microRNA-378a-3p with low expression in Sorafenib-resistant cells Through the establishment of Sorafenib treatment model in tumor-bearing mice combined with qPCR method, the role of miR-378a-3p was further determined The expression level is negatively correlated with the degree of resistance of tumor cells to Sorafenib. Using miR-378a-3p transcription factor LXRα agonist GW3965 on cell drug resistance, IGF1R gene expression and apoptosis, it was determined that GW3965 can promote the transcription of miR-378a-3p and inhibit IGF1R, and reverse tumor cells Resistance to sorafenib. The proposal of the present invention provides a new target for tumor treatment, and provides an effective technical means for effectively combating sorafenib drug resistance.

Description

technical field [0001] The invention belongs to the fields of biotechnology and medicine, in particular, the invention relates to microRNAs related to anti-tumor drug resistance and applications thereof. Background technique [0002] Liver cancer is one of the most common malignant tumors in my country and the world. There are about 500,000 new cases every year in the world, of which hepatocellular carcinoma (HCC) accounts for 85%. my country is a big country with hepatitis B, the incidence of HCC is about 30.1 / 100,000, and the number of new cases and deaths account for more than 50% of the world. Although the 5-year survival rate of HCC has improved with the promotion of tumor markers and imaging examinations, the level of surgery and the development of various new treatment methods such as transcatheter arterial chemoembolization (TACE). But overall, the prognosis of HCC is still unsatisfactory. Surgery is the only cure for HCC patients. In addition, other treatments su...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883C12N15/113A61K45/00A61P35/00A61P1/16
CPCA61K45/00A61P1/16A61P35/00C12Q1/6886C12Q2600/106C12Q2600/158C12Q2600/178
Inventor 徐俊杰蔡秀军林中杰季琳岑栋梁霄姜是夏顺杰陶力野
Owner ZHEJIANG UNIV
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