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Application of circular RNA CircNFIB in diagnosis, treatment and prognosis of intrahepatic cholangiocellular carcinoma

A technology for inner bile duct and cell carcinoma, applied in the prognosis of intrahepatic cholangiocarcinoma, treatment of intrahepatic cholangiocarcinoma, circular RNA CircNFIB in the field of diagnosis, can solve the problem of little research on the role of circular RNA, and achieve inhibition The effect of tumor growth, inhibition of proliferation, and enhancement of anti-tumor activity

Active Publication Date: 2022-02-11
WEST CHINA HOSPITAL SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few studies on the role of circRNAs in ICC metastasis
[0005] Circular RNA cNFIB (CircBase ID: hsa_circ_0086376) contains the 3rd to 6th exons of the gene NFIB, and its circularized nucleotide sequence has 363 bases. At present, there is no diagnosis of circular RNA cNFIB , treatment of intrahepatic cholangiocarcinoma, and application in the prognosis of intrahepatic cholangiocarcinoma

Method used

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  • Application of circular RNA CircNFIB in diagnosis, treatment and prognosis of intrahepatic cholangiocellular carcinoma
  • Application of circular RNA CircNFIB in diagnosis, treatment and prognosis of intrahepatic cholangiocellular carcinoma
  • Application of circular RNA CircNFIB in diagnosis, treatment and prognosis of intrahepatic cholangiocellular carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] In this example, the inventors designed a pair of specific primers capable of amplifying the circular RNA cNFIB, and used the primer pair to amplify cNFIB, and confirmed the splicing site (back-splice site) of the circular RNA by the method of generation sequencing. ), and then through the RNase R digestion experiment, it was determined that cNFIB is a circular RNA molecule with a closed circular structure, and the stability of cNFIB was proved by the actinomycin D experiment.

[0052] The experimental scheme of this embodiment is as follows:

[0053] Total RNA was extracted from RBE cells using Trizol reagent, and the purity and concentration of total cellular RNA was detected by an ultra-micro nucleic acid analyzer. Prepare the reverse transcription reaction system on ice. After the preparation is completed, add the prepared total cellular RNA for reverse transcription. The reverse transcription reaction system is:

[0054]

[0055] The reverse transcription react...

Embodiment 2

[0066] In this example, the expression difference of cNFIB in the tumor tissue and the corresponding non-tumor tissue was detected by fluorescent quantitative PCR, and it was found that the expression of cNFIB in the tumor tissue was significantly higher than that in the corresponding non-tumor tissue, indicating that the expression level of cNFIB can be detected by using for the diagnosis of intrahepatic cholangiocarcinoma.

[0067] The experimental scheme of this embodiment is as follows:

[0068] Total RNA was extracted from cells and tumor tissues using the Total RNA Isolation Kit according to the manufacturer's instructions, using ChamQ TM SYBR@qPCR Master Mix for real-time PCR analysis. Relative expression of RNA was normalized by β-actin or U6 and quantified by the 2-ΔΔCt method.

[0069] Specifically, the cDNA reverse transcription reaction in Example 1 was used to perform tissue RNA reverse transcription.

[0070] The PCR reaction system was prepared on ice (take ...

Embodiment 3

[0085] In this example, using the median value of relative cNFIB expression in ICC tissues, 120 ICC patients were divided into cNFIB low expression group (n=60) and cNFIB high expression group (n=60), β-actin Alternatively U6 was used as an endogenous control.

[0086] like Figure 5 As shown, by Kaplan-Meier survival analysis, ICC patients with lower cNFIB expression levels had shorter overall survival and recurrence-free survival after curative resection.

[0087] Table 1 shows the prognostic factors for overall survival and recurrence-free survival from the univariate COX proportional hazards regression model. In univariate analysis, serum CA19-9 level, tumor number, tumor volume, tumor differentiation, microvascular invasion, lymph node metastasis, TNM stage, and cNFIB expression level were associated with overall survival and recurrence-free survival.

[0088] Table 1:

[0089]

[0090] In multivariate analysis, low expression of cNFIB was associated with poorer tum...

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Abstract

The invention discloses application of circular RNA CircNFIB in diagnosis, treatment and prognosis of intrahepatic cholangiocellular carcinoma. The invention provides a kit for diagnosing the intrahepatic cholangiocellular carcinoma and a pharmaceutical composition for treating the intrahepatic cholangiocellular carcinoma based on the function and mechanism research of the inventor on the circular RNA cNFIB (CircBase ID: hsa_circ_0086376) in growth and metastasis of the intrahepatic cholangiocellular carcinoma. The invention also discloses application of the circular RNA cNFIB as a therapeutic target in prognosis, diagnosis and treatment of intrahepatic cholangiocellular carcinoma.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of circular RNA CircNFIB in the diagnosis and treatment of intrahepatic cholangiocarcinoma and the prognosis of intrahepatic cholangiocarcinoma. Background technique [0002] Intrahepatic cholangiocarcinoma (ICC) is the second most common primary liver malignancy. The incidence of intrahepatic cholangiocarcinoma is increasing worldwide, with an average annual increase of 4.4% over the past decade, due to the increasing prevalence of nonalcoholic fatty liver disease and hepatitis C. ICC is usually asymptomatic in the early stage, and most patients are diagnosed with ICC at an advanced stage, and the available treatment options are limited, resulting in poor clinical outcomes. [0003] Currently, radical resection is still the main means of curing ICC, but 60% of surgical patients have recurrent or metastatic disease. Therefore, there is an urgent need to furt...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12N15/113
CPCC12Q1/6886C12N15/113C12Q2600/118C12Q2600/178C12Q2600/158C12N2310/532Y02A50/30
Inventor 曾勇杜锦鹏黄纪伟兰天袁克非廖皓天冯旭萍陈向征
Owner WEST CHINA HOSPITAL SICHUAN UNIV
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