Specific high-expression UCR sequence in non-small cell lung cancer, detecting kit and detecting method

A non-small cell lung cancer and detection kit technology, applied in the field of tumor molecular biology, to achieve good application prospects, rapid and accurate quantification, and good stability

Active Publication Date: 2018-09-07
YANGZHOU FIRST PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no literature report on the expression of UCR an

Method used

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  • Specific high-expression UCR sequence in non-small cell lung cancer, detecting kit and detecting method
  • Specific high-expression UCR sequence in non-small cell lung cancer, detecting kit and detecting method
  • Specific high-expression UCR sequence in non-small cell lung cancer, detecting kit and detecting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 Analysis of UCR microarray differential expression between non-small cell lung cancer tissue and paired normal tissue

[0058] 1. Materials and methods

[0059] 1. Materials

[0060] The tissue samples were obtained from inpatient surgical resection samples of 20 non-small cell lung cancer patients (numbered 1-20), 15 of which were paired non-small cell lung cancer tissues and adjacent normal tissues.

[0061] 2. Method

[0062] 2.1 Extraction of total RNA from tumor tissue and paired normal tissue

[0063] The total RNA extracted from non-small cell lung cancer tumor tissue and paired normal tissue was operated according to the instructions of Qiagen's RNA extraction kit (RNeasy Micro Kit, #74004). The kit contains RNase-Free DNase I (lyophilized), which can Effectively removes DNA from the genome. The purity and concentration of the extracted RNA were then quantified using a NanoDrop ND 1000 Nucleic Acid Quantifier (NanoDrop Technologies, Wilmington, Del...

Embodiment 2

[0083] Example 2 Preliminary verification of real-time fluorescent quantitative qRT-PCR uc.293 - Differential expression in non-small cell lung cancer tissues and paired adjacent normal tissues

[0084] 1. Experimental materials

[0085] Then select another 40 pairs (numbered 21-60) of non-small cell lung cancer tissues and paired adjacent normal tissues, for uc.293 - The expression difference was verified by qRT-PCR for the first time.

[0086] 2. Experimental methods and results

[0087] 1. Primer specificity screening and identification

[0088] according to uc.293 - Gene loci extracted from UCSC Genome Brower database uc.293 - Related transcript gene sequence, and use Primer 5 primer design software for uc.293 - design primers;

[0089] (1) The designed primers were evaluated by Oligo7, and 3 pairs of designed primer sequences were obtained;

[0090] The first pair: upstream primer SEQ ID NO.2

[0091] Downstream primer SEQ ID NO.3

[0092] The second pair: upstr...

Embodiment 3

[0126] Embodiment 3qRT-PCR further verified uc.293 - Differential expression in tumor tissues and paired normal tissues of non-small cell lung cancer

[0127] 1. qRT-PCR kit composition

[0128] 1.1 dyes uc.293 - PCR kit composition:

[0129] (1) Upstream primer: SEQ ID NO.4

[0130] (2) Downstream primer: SEQ ID NO.5;

[0131] Other reagents refer to SYBR Premix Ex Taq TM II (Tli RNaseH Plus) fluorescence quantitative kit (Code No.RR820A).

[0132] 2.uc.293 - detection

[0133] 2.1 Preparation of total RNA

[0134] Select another 80 pairs of non-small cell lung cancer (numbered 61-130) tumor tissue and paired normal tissue, according to the ThermoFisher Scientific company's Reagents (Product No. 15596018) Reagents and steps required to extract total RNA, please refer to the instructions for details. The purity and concentration of the extracted RNA were then quantified using a NanoDrop ND 1000 Microvolume UV-Vis Spectrophotometer (NanoDrop Technologies, Wilmington...

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Abstract

The invention discloses a specific high-expression UCR sequence in non-small cell lung cancer, a detecting kit and a detecting method. After the UCR expression profile of the non-small cell lung cancer is systemically explored, a set of UCRs related to the invasion and metastasis characteristic of the non-small cell lung cancer is found, and as a result, one UCR, namely, uc.293 remarkably and highexpressed in non-small cell lung cancer tissue is screened out. According to the specific high-expression UCR sequence in the non-small cell lung cancer, the detecting kit and the detecting method, asuperconserved gene sequence 293 related to the specificity of the non-small cell lung cancer is detected, and tumor diagnosis, prognosis and treatment are assisted.

Description

technical field [0001] The invention belongs to the field of tumor molecular biology, and in particular relates to a UCR sequence, a kit and a detection method for detecting highly expressed in non-small cell tissues. Background technique [0002] Non-small cell lung cancer is a common malignant tumor in my country, and its morbidity and mortality are at the forefront of tumors. According to clinicopathological characteristics, it can be divided into non-small cell lung cancer and small cell lung cancer. Among them, non-small cell lung cancer accounts for about 80% of all lung cancers. Most patients are in the middle and advanced stages when they are discovered, and the 5-year survival rate is only 16%. The reasons for treatment failure in patients with non-small cell lung cancer are mainly related to early diagnosis, invasion, metastasis and recurrence of lung cancer cells, which are the main reasons for treatment failure and patient death in non-small cell lung cancer. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12N15/113
CPCC12N15/113C12N2310/14C12Q1/6886C12Q2600/158C12Q2600/178
Inventor 周俊
Owner YANGZHOU FIRST PEOPLES HOSPITAL
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