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Biomarkers for the detection of colorectal cancer and their applications

A technology of colorectal cancer and detection kits, applied in the field of diagnostic markers, can solve problems such as low sensitivity and specificity, poor patient compliance, invasiveness, and high cost, and achieve high specificity, high sensitivity, and simple operation Effect

Active Publication Date: 2022-04-19
江西佰延生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Clinically, the diagnosis of colon adenocarcinoma mainly relies on laboratory tests such as fecal occult blood (FOBT) test, cytological diagnosis, histopathological examination, serum carcinoembryonic antigen (CEA) determination, fiber optic colonoscopy, imaging diagnosis such as colon Air-barium double-contrast radiography, CT scan, MRI, ultrasound section imaging diagnosis, nuclide diagnosis, and colonoscopy are the most reliable methods for the diagnosis of colon adenocarcinoma, but they are invasive, expensive, and poor patient compliance; CT and ultrasound examinations are difficult to detect Small lesions, the early diagnosis of colon adenocarcinoma is limited; CEA is a widely used serum marker in clinical practice, its sensitivity and specificity are low, and it is mainly used for treatment monitoring of patients with colon adenocarcinoma

Method used

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  • Biomarkers for the detection of colorectal cancer and their applications
  • Biomarkers for the detection of colorectal cancer and their applications
  • Biomarkers for the detection of colorectal cancer and their applications

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1 Online big data analysis

[0035] Using online gene analysis data (www.oncomine.org), PAPPA expression was significantly increased in colon adenocarcinoma compared with normal tissue (p=3.29E-11, figure 1 ), the expression was significantly increased in colonic mucinous adenocarcinoma (p=1.95E-7, figure 2 ), the expression was significantly increased in cecum adenocarcinoma (p=9.89E-8, image 3 ), the expression was significantly increased in sigmoid colorectal cancer (p=1.75E-6, Figure 4 ) was significantly increased in rectal adenocarcinoma (p=1.60E-5).

Embodiment 2

[0036] Example 2 The mRNA expression level of PAPPA by qRT-PCR

[0037] 1. Collection of human colon cancer and colitis tissue samples

[0038] Twenty-eight pairs of colorectal cancer tissue specimens and their matched normal tissue specimens, 10 pairs of colitis tissue specimens and their matched normal tissue specimens were collected from the Department of Gastrointestinal Surgery of the First Affiliated Hospital of Jining Medical College, and then the specimens were stored at -80°C.

[0039] 2. RNA extraction

[0040] RNA was extracted from frozen colon cancer and adjacent normal colon tissues using Trizol reagent (Invitrogen, Carlsbad, CA) according to the manual.

[0041] 3. Detection of PAPPA mRNA expression level by qRT-PCR

[0042] Use qRT-PCR to detect the mRNA expression level of PAPPA, reaction system: forward primer (20 μm) 0.15 μl, reverse primer (20 μm) 0.15 μl, cDNA 0.7 μl, H 2 O 9 μl, fluorescent dye SYBR Green (2×) 10 μl; reaction conditions: 50°C for 2 min...

Embodiment 3

[0052] Example 3 Using enzyme-linked immunosorbent assay (ELISA) to detect PAPPA in serum

[0053] 1. Collection of human serum samples

[0054] In this study, a total of 40 serum samples were obtained from Xinxiang Central Hospital, including 20 colon cancer serum samples and 20 normal human serum samples. All colorectal cancer patients were confirmed by pathological examination. All research subjects were fasted for more than 8 hours before blood drawing, and about 5ml of fasting venous blood was collected, left at room temperature for 30 minutes, and then centrifuged at 3000r / min for 10 minutes to collect serum, which was frozen at -20°C for PAPPA concentration monitoring.

[0055] 2. Use enzyme-linked immunosorbent assay (ELISA) to detect the concentration of PAPPA in serum

[0056] Enzyme-linked immunosorbent assay (ELISA) was used to detect the concentration of PAPPA in serum samples. Three repetitions were set for each group of serum samples, and the OD value was dete...

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Abstract

The invention relates to a diagnostic marker, and specifically provides a biomarker for detecting colorectal cancer and its application. The present invention finds through experimental research that the mRNA expression level of pregnancy-associated plasma protein A (PAPPA) gene in colon adenocarcinoma tissue, colonic mucinous adenocarcinoma tissue, cecum adenocarcinoma tissue, sigmoid colorectal cancer tissue and rectal adenocarcinoma tissue and healthy control There were significant differences between them; the expression level of PAPPA protein in the serum of patients with colorectal cancer was also significantly higher than that of normal controls. The present invention proposes the correlation between PAPPA and colorectal cancer for the first time, and verifies through experiments that the available PAPPA can be used to diagnose colorectal cancer, and has the characteristics of simple operation, high specificity and high sensitivity.

Description

technical field [0001] The invention relates to a diagnostic marker, in particular to the application of pregnancy-associated plasma protein A (PAPPA) in detecting colorectal cancer. Background technique [0002] Pregnancy-associated plasma protein A (PAPPA) was originally found in the plasma of pregnant women. During pregnancy, PAPPA is produced by placental syncytiotrophoblast cells and secreted into serum, and its levels are maintained throughout pregnancy. Abnormal circulating PAPPA levels have been associated with fetal disease; for example, low PAPPA levels have been associated with fetal Down syndrome (trisomy 21), preterm birth, low birth weight, and preeclampsia in early pregnancy. In contrast, PAPPA levels were elevated in patients with diabetic nephropathy. [0003] The PAPPA gene is located on chromosome 9q33.1 and encodes a 180 kDa protein of 1627 amino acids. PAPPA plays an important role in bone formation, inflammation and response to injury such as wound h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574C12N15/11
CPCG01N33/57446G01N33/57484C12Q1/6886C12Q2600/158
Inventor 郭永臣鲍永华杨万才
Owner 江西佰延生物技术有限公司
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