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Detection method and application of long-chain non-coded RNA for screening bladder cancer

A technology for bladder cancer and its application, which is applied in the field of tumor molecular biology, can solve the problems of unsatisfactory overall curative effect and poor specificity, and achieve good application prospects, high sensitivity and good specificity

Active Publication Date: 2015-02-18
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The drugs currently used for intravesical infusion to prevent recurrence can reduce the recurrence rate of bladder cancer to a certain extent, but the overall curative effect is not ideal due to poor specificity and the existence of tumor multidrug resistance (MDR).

Method used

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  • Detection method and application of long-chain non-coded RNA for screening bladder cancer
  • Detection method and application of long-chain non-coded RNA for screening bladder cancer
  • Detection method and application of long-chain non-coded RNA for screening bladder cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Analysis of lncRNA chip expression in human bladder cancer and paracancerous tissues

[0048] 1. Materials and methods

[0049] 1. Materials

[0050] Tissue samples were obtained from 3 pairs of inpatient surgical resection samples of bladder cancer patients, each pair containing bladder cancer tissue and paired paracancerous tissue.

[0051] 2. Method

[0052] (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).

[0053] (2) Cy5 fluorescent labeling of sample RNA (entrust Shanghai Kangcheng Bioengineering Co., Ltd. to carry out "ArrayStar Human LncRNA Microarray V3.0Service" for labeling service)

[0054] (3) Synthesis of first-strand cDNA by reverse transcription: start with Total RNA, Oligo(dT)Primer (Shanghai Kangcheng Bioengineering Co., Ltd.) company) to synthesize firs...

Embodiment 2

[0062] Example 2: qRT-PCR preliminary verification of differential expression of lncRNA-AC1 in bladder cancer tissues and paracancerous tissues

[0063] 1. Experimental materials

[0064] 30 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 lncRNA-AC1 by qRT-PCR.

[0065] 2. Experimental methods and results

[0066] 1. Identification of primer specificity

[0067] (1) Design of specific primers: extract lncRNA-AC1-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;

[0068] The designed primer sequences are as follows:

[0069] Upstream primer: SEQ ID No.2

[0070] Downstream primer: SEQ ID No.3

[0071] (2) Extract total RNA from human bladder cancer t...

Embodiment 3

[0112] Example 3: Screening of bladder cancer tissue by differential expression of lncRNA-AC1

[0113] 1. Experimental materials

[0114] 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 lncRNA-AC1 by qRT-PCR.

[0115] 2. Experimental methods and results

[0116] 1. Identification of primer specificity

[0117] (1) Use the following specific primer sequences:

[0118] Upstream primer: SEQ ID No.2

[0119] Downstream primer: SEQ ID No.3

[0120] (2) Extract total RNA from human bladder cancer tissue and paracancerous tissues according to the reagents and steps required by SIGMA's TRIZOL reagent (product number T9424), and then quantify the extracted RNA with a 7300 real time PCR system nucleic acid quantifier (Applied Biosystems AB). RNA purity and concentration.

[0121] (3) Using StarScript II One-step RT-PCR Kit (Product No. A215...

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Abstract

The invention relates to a detection method and application of a long-chain non-coded RNA for screening a bladder cancer. Specifically, an lncRNA which is remarkably and highly expressed in human bladder cancer tissues is screened through variation analysis by utilizing a long-chain non-coded expression profile chip technology, and named as lncRNA-AC1. In comparison with the expression in normal tissues, the lncRNA-AC1 is remarkably and highly expressed in the human bladder cancer tissues, and a large sample fluorescent quantitative PCR experiment further proves that the expression quantity of the lncRNA-AC1 in the human bladder cancer tissues is obviously higher than that in the normal tissues. Researches on pathogenesis of the bladder cancer can be further enriched by the new lncRAN-AC1, and a new tumor marker and a new therapeutic target are provided for early diagnosis and prognosis monitoring of the bladder cancer.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to a long-chain non-coding RNA and its application. Specifically, the present invention relates to the application of a long-chain non-coding RNA in preparing preparations for auxiliary diagnosis or prognosis of bladder cancer. Background technique [0002] Bladder cancer is the most common malignant tumor of the urinary system, and its incidence rate is listed as the fourth among male malignancies and the seventh among female malignancies in the United States, second only to prostate cancer. In my country, the incidence rate is the eighth, ranking first in urinary system tumors. About 95% of bladder cancer originates from the bladder epithelium, most of which are malignant, among which transitional cell carcinoma is the most common, accounting for more than 80%. "Prevention of recurrence" of bladder cancer after tumor resection is an important clinical problem. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 范祖森李翀杜颖杨昭何璐云王彦英阎新龙朱平平
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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