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LOC100128675 serving as molecular marker for detecting prostate cancer and application of molecular marker to diagnostic kit

A technology of molecular markers and diagnostic kits, applied in the field of molecular markers and their application in diagnostic kits, can solve the problems of undiscovered prostate cancer related reports, and achieve the effect of rapid and effective early detection

Inactive Publication Date: 2017-02-01
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

[0004] LOC100128675, also known as HPN-AS1, belongs to lncRNA (longnon-coding RNA, long-chain non-coding RNA), located on chromosome 19, is a type of non-coding RNA with a transcript length of more than 200 bp nucleotides, in liver, pancreas, kidney It is overexpressed in the cortex, and there is no report about it in prostate cancer

Method used

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  • LOC100128675 serving as molecular marker for detecting prostate cancer and application of molecular marker to diagnostic kit
  • LOC100128675 serving as molecular marker for detecting prostate cancer and application of molecular marker to diagnostic kit
  • LOC100128675 serving as molecular marker for detecting prostate cancer and application of molecular marker to diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 High-throughput sequencing screening of differentially expressed genes

[0038] 1. Sampling

[0039] Select 10 cases of fresh prostate cancer tissues and corresponding paracancerous tissues, samples from prostate cancer patients who underwent radical prostatectomy and bilateral pelvic lymph node dissection from October 2012 to December 2015 in Peking Union Medical College Hospital. Under the guidance of the pathologist, the prostate cancer and paracancerous tissues were immediately taken and put into liquid nitrogen, and stored in a -80°C low-temperature refrigerator after serial numbering.

[0040] 2. Total RNA extraction from tissue samples

[0041] use Reagent (invitrogen, Cat. No. 15596-018) was used to extract sample RNA, and the experimental operation was carried out according to the product manual. The specific operation was as follows:

[0042] Collect the samples and freeze them in liquid nitrogen. After taking them out, put the tissue samples in...

Embodiment 2

[0053] Example 2 RT-PCR verification of the expression of LOC100128675 in prostate cancer tissue

[0054] 1. Materials

[0055] 10 prostate cancer patients were selected, and the samples were collected from the prostate cancer patients who underwent radical prostatectomy and bilateral pelvic lymph node dissection from October 2012 to December 2015 in Peking Union Medical College Hospital. Immediately take prostate cancer and paracancerous tissues and put them into liquid nitrogen, and store them in a -80°C low-temperature refrigerator after serial numbering.

[0056] 2. Method

[0057] 2.1 Extract the total RNA from the tissue samples of the subjects, the same as the extraction method in Example 1.

[0058] 2.2 Synthesis of cDNA by reverse transcription

[0059] use III Reverse Transcriptase (invitrogen, Cat. No. 18080-044) was used for cDNA reverse transcription, and the experimental operation was carried out according to the product manual. The specific operation was a...

Embodiment 3

[0081] The preparation of embodiment 3 kits

[0082] Based on the primer set obtained in Example 2, assemble the prostate cancer diagnostic kit of the present invention, specifically amplify the primer pair of the nucleotide sequence shown in SEQ ID NO:2 such as SEQ ID NO:3 and SEQ ID NO Shown in: 4, and the primer pair of specific amplification housekeeping gene (GAPDH) as shown in SEQ ID NO: 5 and SEQ ID NO: 6; Also comprise SYBR Green polymerase chain reaction system, as PCR damping fluid, SYBR Green Fluorescent dyes, dNTPs. The composition of described PCR buffer is 25mMKCl, 2.5mM MgCl 2 , 200mM (NH 4 ) 2 SO 4 .

[0083] Through the optimization of primer concentration and annealing temperature, the composition of the prostate cancer diagnostic kit is finally determined as follows:

[0084](1) Total RNA extraction reagents from tissue samples, blood or urine:

[0085] Use per 50 mg of tissue or 200 μL of blood or 200 μL of urine:

[0086]

[0087]

[0088] (2...

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Abstract

The invention discloses a molecular marker related to prostate cancer, wherein the molecular marker is LOC100128675. The invention further discloses a diagnostic kit for the prostate cancer and an application of the molecular marker to screening, diagnosis, therapy, monitoring and prognosis for prostate cancer high-risk groups. The diagnostic kit comprises the molecular marker LOC100128675 related to the prostate cancer. By the aid of the molecular marker and the diagnostic kit comprising the molecular marker for detecting the prostate cancer, early detection can be rapidly and effectively realized, and therapeutic targets and important basis are provided for clinical application of gene therapy, drug therapy and the like.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to LOC100128675 as a molecular marker for detecting prostate cancer and its application in a diagnostic kit. Background technique [0002] Prostate cancer is a malignant tumor that occurs in the prostate tissue of men. It has become the second most common tumor in men worldwide and ranks sixth in cancer-related mortality. In developed countries and regions such as Europe and the United States, it is the most common malignant tumor in men, and its mortality rate ranks second among various cancers; in Asia, its incidence rate is lower than that of Western countries, but it has shown a rapid upward trend in recent years faster than developed countries in Europe and America. The incidence of prostate cancer in my country is also increasing year by year, ranking third among male genitourinary system malignancies, second only to bladder cancer and kidney cancer. Prostate cancer generally oc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113A61K45/00A61P35/00C12N15/11
CPCA61K45/00C12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178
Inventor 刘昊
Owner 北京致成生物医学科技有限公司
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