Cyclic RNAcircMDK, application thereof and liver cancer treatment preparation

A liver cancer and circular technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, medical preparations of non-active ingredients, etc., can solve the problem of no relevant research reports on functions, achieve true and reliable results, rigorous test process, and easy operation convenient effect

Pending Publication Date: 2022-07-01
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Studies have shown that siRNA (including lncRNA-siRNA and mRNA-siRNA) delivered by nanomaterials can play an anti-tumor role in the treatme

Method used

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  • Cyclic RNAcircMDK, application thereof and liver cancer treatment preparation
  • Cyclic RNAcircMDK, application thereof and liver cancer treatment preparation
  • Cyclic RNAcircMDK, application thereof and liver cancer treatment preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: Screening of differentially expressed circRNAs by RNA-seq experiment

[0055] RNA-Seq sequencing was performed on liver cancer tissues to screen for differentially expressed circRNAs. This part of the experiment was completed by Guangzhou Ribo Biotechnology Co., Ltd. The specific steps are as follows: after RNA extraction and quality inspection, rRNA is removed by probe, and RNaseR delinearizes RNA to construct Illumina sequencing library. Sequencing was then performed, the sequencing mode was PE150, and the sequencing data volume was 12G / 15G. For the sequencing data, first perform Raw Data filtering, de-linker sequences and low-quality reads to obtain high-quality data (Clean Data). The ribosomal RNA sequences were then removed by aligning with the ribosomal database to obtain valid reads. The effective reads are compared with the reference genome, and the comparison results are used for the identification of circular RNAs in the next step. RiboBio uses tw...

Embodiment 2

[0060] Example 2: qPCR detection of circular RNA circMDK expression in liver cancer tissue

[0061] 35 pairs of fresh liver cancer and adjacent tissues were collected to detect the expression of circular RNA circMDK by qPCR. The results are as follows image 3 shown. Circular RNA circMDK is highly expressed in cancer tissues and low in paracancerous tissues.

[0062] The expression levels of circular RNA circMDK in human normal hepatocytes LO2 and hepatoma cells HepG2, Huh7, and Hep3B were detected by qPCR. like Figure 4 As shown, the expression level of circular RNA circMDK was significantly up-regulated in Hep3B, Huh7, and HepG2 cells compared with LO2 cells.

Embodiment 3

[0063] Example 3: RNase R and Actinomycin D treatment and in situ hybridization experiments to detect the structural characteristics of circular RNA circMDK

[0064] RNase R is a highly processed 3' to 5' exoribonuclease that digests linear RNA (i.e. MDK), and resistance experiments to its digestion confirmed that circMDK has a closed loop structure ( Figure 5 ); in addition, after actinomycin D treatment, the half-life of circMDK transcripts within 24 h was more stable than that of MDK ( Image 6 ). The results of nuclear and cytoplasmic separation and RNAFISH detection together showed that circMDK was mainly localized in the cytoplasm ( Figure 7 , Figure 8 ).

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Abstract

The invention discloses a circular RNAcircMDK (Ribonucleic Acid CircMDK), application thereof and a liver cancer treatment preparation. Researches prove that circMDK is a novel circular RNA and is highly expressed in liver cancer tissues, and knock-down of circMDK (siRNA) can inhibit proliferation, migration and invasion of liver cancer cells and promote apoptosis. According to the invention, poly (beta-amino ester) (PAEs) cationic nanoparticles are prepared by adopting a single-step nano precipitation method, and efficient encapsulation is carried out on siRNA. The PAE-siRNA compound down-regulates the expression of circMDK through siRNA silencing, effectively inhibits the occurrence and progress of liver cancer, and has profound clinical significance and application prospects in targeted therapy of liver cancer.

Description

technical field [0001] The invention belongs to the technical field of tumor molecular biology, and in particular relates to the application of a circular RNA circMDK, a reagent for detecting the expression of circMDK in the preparation of a diagnostic preparation for liver cancer, the application of a reagent for inhibiting the expression of circMDK in the preparation of a therapeutic preparation for liver cancer, and a novel nanoparticle polymer (β-aminoester) liver cancer therapeutic preparations carrying circMDK siRNA. Background technique [0002] Liver cancer is the sixth most common cancer worldwide and the second leading cause of cancer-related death. The disease poses a serious threat to health and imposes a heavy economic burden on society and individuals. Common treatment methods for liver cancer include surgery, radiotherapy, chemotherapy, etc. However, conventional treatment methods have large side effects, and often fail to achieve good results for patients wh...

Claims

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Application Information

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IPC IPC(8): C07H21/02A61K31/7088A61K9/51A61K47/34A61P35/00A61P35/04C12Q1/6886C12Q1/6851C12N15/11
CPCC07H21/02A61K31/7088A61K9/5153A61P35/00A61P35/04C12Q1/6886C12Q1/6851C12Q2600/178C12Q2600/158C12Q2600/166C12Q2525/307C12Q2525/207C12Q2561/113C12Q2531/113C12Q2521/107Y02A50/30
Inventor 夏赞贤杜阿帅李诗琴
Owner CENT SOUTH UNIV
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