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Marker for detecting prostate

A prostate cancer and marker technology, applied in the field of biomedicine, can solve problems such as affecting the effect of tumor treatment

Inactive Publication Date: 2016-11-23
北京致成生物医学科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chemoradiotherapy drugs not only act on the tumor, but can also act on the healthy tissues around the tumor. Therefore, while killing the tumor, they also bring great side effects to the body, which ultimately affects the therapeutic effect on the tumor.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 High-throughput sequencing and screening of differentially expressed genes

[0043] 1. Sampling

[0044] Twenty-seven tissue specimens obtained during the urological surgery of Peking Union Medical College Hospital from October 2011 to December 2015 were collected. All specimens were confirmed by pathological examination, including 8 samples of adjacent tissues and 19 specimens of prostate cancer. Store in a low temperature refrigerator at -80℃.

[0045] 2. Extract total RNA from tissue samples

[0046] use Reagent (invitrogen, article number 15596-018) is used to extract RNA from the sample. The experimental operation is carried out according to the product manual. The specific operation is as follows:

[0047] Collect the sample and freeze it in liquid nitrogen. After taking it out, place the tissue in a pre-cooled mortar for grinding. After the tissue sample is powdered:

[0048] ①Add Trizol and store at room temperature for 5 minutes;

[0049] ②Add 0.2 mL of chlorof...

Embodiment 2

[0058] Example 2 RT-PCR verification of gene expression in prostate cancer tissues and adjacent tissues

[0059] 1. Material

[0060] Thirty-four prostate cancer tissue samples and 8 adjacent tissue samples were collected from prostate cancer samples during urological operations of Peking Union Medical College Hospital from 2011 to 2015, and they were grouped and numbered. All samples were confirmed by pathological examination.

[0061] 2. Method

[0062] 2.1 Extract total RNA from tissue samples, the same as the extraction method in Example 1.

[0063] 2.2 Reverse transcription synthesis of cDNA

[0064] use III Reverse Transcriptase (invitrogen, article number 18080-044) was used for cDNA reverse transcription. The experimental operation was carried out according to the product manual. The specific operation is as follows:

[0065] Using reverse transcription kit, reverse transcription of lμg total RNA with reverse transcription buffer to synthesize cDNA. A 25μL reaction system was u...

Embodiment 3

[0085] Example 3 Preparation of a kit for prognosis assessment of prostate cancer

[0086] Based on the primer set obtained in Example 2, the kit for detecting prostate cancer of the present invention was assembled, and the kit includes a primer set for specifically amplifying one or more genes in PPIEL, RAET1K and / or MBL1P, as shown in the table As shown in 1, specifically:

[0087] 1. The kit includes specific amplification PPIEL and RAET1K;

[0088] 2. The kit includes specific amplification PPIEL, RAET1K and MBL1P.

[0089] The primer pair for specific amplification of housekeeping gene (GAPDH) is shown in SEQ ID NO: 7 and SEQ ID NO: 8; it also includes SYBR Green polymerase chain reaction system, such as PCR buffer, SYBR Green fluorescent dye, dNTPs. The composition of the PCR buffer is 25mM KCl, 2.5mM MgCl 2 , 200mM(NH 4 ) 2 SO 4 ; Also includes normal prostate tissue cDNA: as a negative control and the test sample cDNA together for quantitative PCR detection, each reaction sys...

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Abstract

The invention discloses a marker for detecting prostate. The marker is selected from one or more genes in PPIEL, RAET1K and / or MBL1P. The invention further discloses application of the marker in preparation of products of diagnosis and prognosis of prostate. The invention further discloses a kit for the diagnosis or the prognosis evaluation of the prostate. The kit comprises one or multiple groups of primer sequences in specifically amplified PPIEL, RAET1K and / or MBL1P. According to the marker, one or more genes are utilized as the marker for the combined detection of the marker, so that the early detection and the prognosis evaluation can be rapidly and efficiently realized, so that the accuracy is greatly improved, and furthermore, a therapeutic target and the important basis are provided for the clinical application including gene therapy, drug therapy and the like.

Description

Technical field [0001] The present invention relates to the field of biomedicine, in particular to the application of the expression level of prostate cancer-related genes to diagnose cancer and monitor the therapeutic effect. Background technique [0002] Prostate cancer is one of the common malignant tumors of the male reproductive system. Worldwide, the incidence of prostate cancer ranks second among all malignant tumors in men. In the United States, the incidence of prostate cancer has exceeded that of lung cancer, becoming the first tumor that endangers men’s health. The incidence of prostate cancer in Asia is far lower than that in European and American countries, but it has shown an upward trend in recent years, and it has grown more rapidly than in developed countries in Europe and America. Prostate cancer patients are mainly elderly men, usually after the age of 50, 95% of them occur in elderly men over 60, and the incidence continues to increase with age. Prostate can...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/158
Inventor 刘昊
Owner 北京致成生物医学科技有限公司
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