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Prostatic cancer markers from prostatic secretion

A prostate cancer and prostatic fluid technology, applied in the fields of biotechnology and clinical diagnosis, can solve the problems of waste of medical resources and costs, ineffective results, low specificity and sensitivity of clinical diagnosis, etc.

Inactive Publication Date: 2013-03-27
SHANGHAI INST OF PLANNED PARENTHOOD RES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] At present, the main diagnostic marker for the diagnosis of prostate cancer (PCa) is PSA, but its specificity and sensitivity are not high in clinical diagnosis, resulting in a large number of unnecessary biopsies in clinical practice
These problems bring many side effects to patients on the one hand, and cause a lot of waste of medical resources and expenses on the other hand
Currently, the dynamic range of proteins that can be detected by mass spectrometry is only 10 2 -10 4 , far from enough, even if the protein antibody column is used to remove high-abundance proteins, the effect is still not obvious

Method used

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  • Prostatic cancer markers from prostatic secretion
  • Prostatic cancer markers from prostatic secretion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1, collection and processing of prostatic fluid

[0055] 1.1 Add 5-10ul, 300 times working solubility, 3 kinds of proteolytic enzyme inhibitor mixture (working solubility is as follows: Leupepetin 5ug / ml, Antipain 5ug / ml and Chymo statin 10ug / ml) to 1.5ml Eppendorf tube.

[0056] After aliquoting, store the tubes in a -20°C refrigerator for later use, or use them directly in the next steps.

[0057] 1.2 Use conventional clinical techniques to collect prostate fluid and drop it into the tube of 1.1 (about 50ul).

[0058] 1.3 Mix briefly (avoid Votexing). The following steps are all carried out at low temperature.

[0059] 1.4 centrifugation. 3000rpm / min, 5-10 minutes.

[0060] 1.5 Collect the supernatant and transfer it to another pre-cooled Eppendorf tube, this part is the prostate secretion (EPS). When transferring the liquid, do not disturb the sedimented fraction.

[0061] 1.7 All samples above should be stored in -80°C refrigerator.

[0062] 1.8 Ma...

Embodiment 2

[0063] Embodiment 2, the proteomics research method of sample

[0064] 2.1 EPS sample processing

[0065] 2.1.1 Quantitatively determine the protein concentration of each sample by conventional methods.

[0066] 2.1.2 Mix 20 EPS samples of prostate cancer patients and non-cancer patients of the same age in equal amounts (20ug protein / part), that is, the samples of the experimental group (30 samples of prostate cancer patients) and the control Group samples (mixture of 30 samples from non-cancer patients).

[0067] 2.1.2 Add trichloroacetic acid to the sample with a final concentration of 50% (weight / volume) to precipitate proteins and achieve the purpose of moderately purifying proteins. The samples were centrifuged at 5000 rpm / min for 5 minutes, and the supernatant was removed. Take the routine sample hydration buffer stored at -20°C from the refrigerator, and dissolve it at room temperature. Add a total volume of 250ul sample hydration buffer to each group of samples and...

Embodiment 3

[0083] Example 3, Detection of Hepatoglobin Levels in Prostate Secreted Fluid of Prostate Cancer Patient Group and Non-cancer Control Group

[0084] Prostate cancer patient group and non-cancer control group were randomly divided into three copies of prostate secretion fluid, and 10 micrograms of protein in each copy were used for Western Blotting analysis. The rabbit polyclonal antibody (primary antibody) of Hepatoglobin is the product of AbCam Company (ab85846), and the dilution factor is 3000-5000 times when used. In addition, other details involved in the Western Blotting analysis of prostatic fluid are conventional techniques. figure 2 Shows the results of Western Blotting analysis of prostatic fluid.

[0085] According to literature reports, the full-length human Hepatoglobin protein consists of 406 amino acids. Among them, the leader peptide consists of 18 amino acids, and the alpha chain consists of 144 amino acids. The beta chain consists of 244 amino acids. The ...

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Abstract

The invention relates to prostatic cancer markers from a prostatic secretion, and discloses a group of new prostatic cancer markers comprising Haptoglobin alpha-chains, Fibronectin1, Albumin and / or SERPINB1. In the prostatic secretion, the expression of the above proteins in prostatic cancer patients is substantially higher than the expression in non-cancer patients, so reagents or kits for the diagnosis, the assessment and the prognosis of the prostatic cancer can be developed based on the proteins.

Description

technical field [0001] The invention belongs to the fields of biotechnology and clinical diagnosis; more specifically, the invention relates to a prostate cancer marker derived from prostate secretion. Background technique [0002] At present, the main diagnostic marker for the diagnosis of prostate cancer (PCa) is PSA, but its specificity and sensitivity are not high in clinical diagnosis, which leads to a large number of unnecessary biopsies in clinical practice. These problems bring many side effects to patients on the one hand, and cause a lot of waste of medical resources and expenses on the other hand. [0003] Early diagnosis of PCa, selective treatment or simple follow-up observation of confirmed PCa, and avoiding overtreatment are the keys to the successful diagnosis and treatment of PCa. At present, clinical methods mainly rely on PSA detection, digital rectal examination, imaging examination and histopathological examination, but they all have certain limitations...

Claims

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Application Information

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IPC IPC(8): G01N33/574
Inventor 李润生林标扬王健刘媛媛
Owner SHANGHAI INST OF PLANNED PARENTHOOD RES
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