Method for detecting circular RNA for bladder cancer screening and application thereof

A technology of bladder cancer and circle, applied in the field of tumor molecular biology, can solve the problems of unsatisfactory curative effect of bladder cancer and achieve good application prospects, high sensitivity and good specificity

Pending Publication Date: 2016-10-12
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Drugs currently used to prevent bladder cancer recurrence are suboptimal

Method used

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  • Method for detecting circular RNA for bladder cancer screening and application thereof
  • Method for detecting circular RNA for bladder cancer screening and application thereof
  • Method for detecting circular RNA for bladder cancer screening and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Example 1: circRNA chip expression analysis of human bladder cancer and paracancerous tissues

[0045] 1. Materials and methods

[0046] 1. Materials

[0047] The tissue samples were obtained from surgical resection samples of 3 pairs of inpatients with bladder cancer (all tissue samples were obtained from the Urology Department of the Second Affiliated Hospital of Kunming Medical University), and each pair contained bladder cancer tissue and paired paracancerous tissue.

[0048] 2. Method

[0049] (1) Extraction of total RNA from tumor tissue and normal tissue: Total RNA from bladder cancer tissue and paracancerous tissue was extracted according to the instructions of Qiagen's RNA extraction kit (RNeasy Micro Kit, Cat. No. 74004).

[0050] (2) Cy5 fluorescent labeling of sample RNA (Shanghai Kangcheng Bioengineering Co., Ltd. is entrusted with "ArrayStar Human CircRNA Microarray V3.0Service" for labeling service)

[0051](3) Synthesis of first-strand cDNA by reverse...

Embodiment 2

[0059] Example 2: qRT-PCR preliminary verification of differential expression of cRNA-ZFR in bladder cancer tissues and paracancerous tissues

[0060] 1. Experimental materials

[0061] 35 pairs (different from the samples tested by the microarray) of human bladder cancer tissues (provided by the Second Affiliated Hospital of Kunming Medical University) and paired paracancerous tissues were selected to verify the expression difference of cRNA-ZFR by qRT-PCR.

[0062] 2. Experimental methods and results

[0063] 1. Identification of primer specificity

[0064] (1) Design of specific primers: extract cRNA-ZFR-related transcript sequences from the Ensemble database, and use the primer design tool (Primer BLAST) of NCBI to design primers according to the sequence of the transcript;

[0065] The designed primer sequences are as follows:

[0066] Upstream primer: SEQ ID No.2

[0067] Downstream primer: SEQ ID No.3

[0068] (2) Extract total RNA from human bladder cancer tissues...

Embodiment 3

[0105] Example 3: Utilizing differential expression of cRNA-ZFR to screen bladder cancer tissue

[0106] 1. Experimental materials

[0107] 100 human bladder cancer tissues and 100 paracancerous tissues (provided by the Second Affiliated Hospital of Kunming Medical University) were selected to detect the expression difference of cRNA-ZFR by qRT-PCR.

[0108] 2. Experimental methods and results

[0109] 1. Identification of primer specificity

[0110] (1) Use the following specific primer sequences:

[0111] Upstream primer: SEQ ID No.2

[0112] Downstream primer: SEQ ID No.3

[0113] (2) Extract total RNA from human bladder cancer tissues and paracancerous tissues according to the reagents and steps required by SIGMA's TRIZOL reagent (product number T9424), and then quantify the extracted RNA using a 7300 real-time PCR system nucleic acid quantifier (Applied Biosystems AB). RNA purity and concentration.

[0114] (3) Using StarScript II One-step RT-PCR Kit (Product No. A215...

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Abstract

The invention relates to a method for detecting a circular RNA for bladder cancer screening and an application thereof; in particular, with use of a circular RNA chip, through difference analysis, the circRNA significantly highly expressed in a human bladder cancer tissue is screened and is named as cRNA-ZFR. Compared with a normal tissue, the cRNA-ZFR is significantly highly expressed in the human bladder cancer tissue, and the cRNA-ZFR is further confirmed to have the expression quantity in the human bladder cancer tissue significantly higher than that of the normal tissue in large-sample fluorescent quantitative PCR experiments. The novel cRNA-ZFR provides a new idea for further study of pathogenesis of bladder cancer, and also provides a new target for early screening and prognosis monitoring of bladder cancer.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to a circular RNA and its application. Specifically, the present invention relates to the application of a circular RNA in preparing preparations for auxiliary diagnosis or prognosis monitoring of bladder cancer. Background technique [0002] Bladder cancer is the most common malignant tumor in the urinary system, about 95% of which originate from the bladder epithelium. The efficacy of drugs currently used to prevent recurrence of bladder cancer is not satisfactory. Up to 40%-80% of patients will experience one or more recurrences, and 10%-15% of patients will develop higher-grade tumors or develop metastases. Therefore, it is particularly important to discover new bladder tumor markers for early and effective detection of tumor occurrence, development and prognosis monitoring. Circular RNA (circRNA) is a closed circular non-coding RNA molecule composed of multi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 李翀王剑松李书成郝俊峰田勇韩玉刚张勇牛海涛
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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