cRNA-FUT8 detection applied to hepatocellular carcinoma screening and application thereof

A technology for hepatocellular carcinoma, which is applied in the field of circular RNA, can solve the problems of no treatment method and limited surgical efficacy, and achieve the effect of high sensitivity, good specificity and good application prospects

Inactive Publication Date: 2016-07-27
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At this time, in addition to surgery, there is often no treatment available, and the curative effect of surgery is very limited, and the median survival time is only a few months

Method used

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  • cRNA-FUT8 detection applied to hepatocellular carcinoma screening and application thereof
  • cRNA-FUT8 detection applied to hepatocellular carcinoma screening and application thereof
  • cRNA-FUT8 detection applied to hepatocellular carcinoma screening and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Analysis of cRNA-FUT8 chip expression in human hepatocellular carcinoma and paracancerous tissues

[0047] 1. Materials and methods

[0048] 1. Materials

[0049] Tissue samples were obtained from surgical resection samples of 3 pairs of inpatients with HCC (all tissue samples were obtained from the Department of Liver Tumor Surgery, Shanghai Zhongshan Hospital), and each pair contained HCC tissue and paired adjacent normal tissue.

[0050] 2. Method

[0051] (1) Extraction of total RNA from tumor tissue and normal tissue: Total RNA from hepatocellular carcinoma tissue and paracancerous tissue was extracted according to the instructions of Qiagen's RNA extraction kit (RNeasy Micro Kit, Cat. No. 74004).

[0052] (2) Cy5 fluorescent labeling of sample RNA (Shanghai Kangcheng Bioengineering Co., Ltd. is entrusted with "ArrayStarHumancRNA-FUT8MicroarrayV3.0Service" for labeling service)

[0053] (3) Synthesis of first-strand cDNA by reverse transcription: star...

Embodiment 2

[0061] Example 2: qRT-PCR preliminary verification of differential expression of cRNA-FUT8 in cancer tissues and paracancerous tissues of hepatocellular carcinoma

[0062] 1. Experimental materials

[0063] 28 pairs (different from the samples tested by the microarray) of human hepatocellular carcinoma tissues (provided by the Department of Liver Tumor Surgery, Shanghai Zhongshan Hospital) and paired paracancerous tissues were selected to verify the expression difference of cRNA-FUT8 by qRT-PCR.

[0064] 2. Experimental methods and results

[0065] 1. Identification of primer specificity

[0066] (1) Design of specific primers: extract the transcript sequence related to cRNA-FUT8 from the Ensemble database, and use the primer design tool (PrimerBLAST) of NCBI to design primers according to the sequence of the transcript;

[0067] The designed primer sequences are as follows:

[0068] Upstream primer: SEQIDNo.2

[0069] Downstream primer: SEQIDNo.3

[0070] (2) Extract tot...

Embodiment 3

[0109] Example 3: Using differential expression of cRNA-FUT8 to screen hepatocellular carcinoma tissues

[0110] 1. Experimental materials

[0111] 100 human hepatocellular carcinoma tissues and 100 para-cancerous tissues (provided by Shanghai Renji Hospital) were selected to detect the expression difference of cRNA-FUT8 by qRT-PCR.

[0112] 2. Experimental methods and results

[0113] 1. Identification of primer specificity

[0114] (1) Use the following specific primer sequences:

[0115] Upstream primer: SEQIDNo.2

[0116] Downstream primer: SEQIDNo.3

[0117] (2) Extract total RNA from human hepatocellular carcinoma tissue and paracancerous tissue according to the reagents and steps required by SIGMA's TRIZOL reagent (product number T9424), and then quantify the purity and concentration.

[0118] (3) The extracted total RNA was reverse-transcribed to synthesize cDNA using StarScript II One-step RT-PCR Kit (Product No. A215-01) from Beijing Kangrun Chengye Biotechnolo...

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Abstract

The invention mainly relates to cRNA-FUT8 detection applied to hepatocellular carcinoma screening and application thereof. A specific method comprises the steps that a gene chip technique is utilized to carry out cRNA-FUT8 expression profile differential analysis on carcinoma para-carcinoma tissue cells to screen out an obvious high-expression circular RNA, cRNA-FUT8, in a hepatocellular carcinoma tissue of a person. Compared with a para-carcinoma normal tissue, the circular RNA has obvious high expression in the hepatocellular carcinoma tissue of the person. Furthermore, a real-time quantitative PCR, Northern Blot experiment further proves that the expression quantity of the cRNA-FUT8 in the hepatocellular carcinoma tissue of the person is obviously higher than that in the normal tissue. The new-researched cRNA-FUT8 can provide a novel view for a research on a relapsing and metastasis mechanism of the hepatocellular carcinoma and also provides a new evaluating index and a new intervening target spot for diagnosing, preventing and curing the hepatocellular carcinoma.

Description

technical field [0001] The present invention belongs to the field of tumor molecular biology, and is mainly used for the detection and application of a circular RNA. Specifically, the present invention relates to a circular RNA, that is, cRNA-FUT8, which is used in the preparation of auxiliary diagnosis, prognosis evaluation and even Application in related preparations for the treatment of hepatocellular carcinoma. Background technique [0002] Hepatocellular carcinoma (HCC) (referred to as liver cancer) is one of the most common malignant tumors in the world. Among the causes of cancer death in men, liver cancer ranks second, surpassing gastric cancer and second only to lung cancer, and its incidence is steadily increasing , while the number of liver cancer patients in my country accounts for half of the global total. Liver cancer has the problems of "difficult early diagnosis", "high recurrence rate", and "poor prognosis". Its onset is rapid and hidden, and most patients w...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
CPCC12N15/11C12N2310/10C12Q1/6837C12Q1/6851C12Q1/6886C12Q2600/106C12Q2600/118C12Q2525/307C12Q2531/113C12Q2563/107
Inventor 李翀郝俊峰李书成孟姝田勇韩玉刚林开粟朱虹
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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