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Tumor markers associated with pancreatic cancer and application thereof

A pancreatic cancer marker technology, applied in the fields of genetic engineering and oncology, can solve the problems of low sensitivity, inaccurate and elevated pancreatic cancer indicators

Active Publication Date: 2014-03-26
THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These markers are imprecise and less sensitive as indicators for the diagnosis of pancreatic cancer alone, and are also elevated in other gastrointestinal tumors and benign diseases, so the value of early diagnosis of pancreatic cancer is limited
In addition, some genetic tumor markers are also used in the early diagnosis of clinical pancreatic cancer, but the purpose of diagnosis has not been achieved

Method used

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  • Tumor markers associated with pancreatic cancer and application thereof
  • Tumor markers associated with pancreatic cancer and application thereof
  • Tumor markers associated with pancreatic cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1 sample collection and processing

[0055] Tissue samples of 3 patients with pancreatic cancer and 3 cases of control group were clinically collected, and the clinical data of the patients were collected systematically.

Embodiment 2

[0056] Roche NimbleGen Chip Detection of mRNA in Example 2 Pancreatic Cancer and Normal Tissue

[0057] 3 cases of pancreatic cancer patients and 3 cases of healthy controls were tested by Roche NimbleGen chip to obtain relevant results. The specific steps are:

[0058] 1. Take the pancreatic tissues of the "pancreatic cancer case" group and the "healthy control" group respectively, add Trizol reagent, and use the chloroform-isopropanol method to extract total RNA;

[0059] 2. Quantitatively measure total RNA concentration by spectrophotometer, and detect its integrity by agarose gel electrophoresis;

[0060] 3. Reverse transcription to synthesize single-stranded cDNA. The reverse transcription reaction system includes 5 μL of single-strand synthesis Mix (4 μL of single-strand buffer Mix, 1 μL of single-strand synthetase), 5 μl (0.2 μg) of total RNA, and the system is incubated at 42°C for 2 hours;

[0061] 4. Two-strand cDNA synthesis. Prepared on ice, the reaction system...

Embodiment 3

[0067] CST1, CST4, CST2mRNA qRT-PCR experiment in the tissue of embodiment 3

[0068] 1. According to the results of the gene chip, the differentially expressed genes CST1, CST4, and CST2 were selected for further verification by qRT-PCR.

[0069] 2. Extraction of total RNA from tissue samples, using the chloroform extraction method to extract total RNA from tissue samples, the specific steps are as follows:

[0070] (1) Take the frozen pancreas tissue sample from the patient, knock out a tissue block with a size of about 2mm×2mm×2mm, add 1ml Trizol reagent to lyse the tissue, and repeatedly blow it with a gun or shake vigorously to lyse the tissue;

[0071] (2) Transfer the Trizol lysate of the above tissue into an EP tube and place it at room temperature 15-30°C for 5 minutes;

[0072] (3) In the above EP tube, add 0.2ml chloroform for every 1ml TRIZOL, cover the EP tube cap, vibrate vigorously in your hands for 15 seconds, and place it at room temperature (15°C-30°C) for 2...

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Abstract

The invention belongs to the field of genetic engineering and medicine, relates to a novel tumor marker for early diagnosis of pancreatic cancer, and more specifically discloses a new serum / tissue tumor marker associated with pancreatic cancer and application thereof. The marker is a combination of CystatinSN (CST1), CystatinS (CST4) and CystatinSA (CST2). The marker, primers and antibodies can be used for a diagnostic kit for the auxiliary diagnosis of pancreatic cancer.

Description

technical field [0001] The invention belongs to the fields of genetic engineering and oncology, and relates to a serum / tissue tumor marker related to pancreatic cancer and its application. Background technique [0002] Pancreatic cancer is a tumor with a high degree of malignancy and a very poor prognosis, and its incidence is increasing year by year worldwide. According to data from the American Cancer Society, the incidence of pancreatic cancer ranks 10th among malignant tumors, and its case fatality rate ranks 4th among malignant tumors. The 5-year survival rate is only 5%. Due to the deep anatomical location of the pancreas, clinically specific symptoms such as abdominal pain, weight loss, and jaundice are difficult to detect, and most patients have already developed cancer metastasis when they are diagnosed. Therefore, early diagnosis is a key factor in improving the cure rate of pancreatic cancer, and finding tumor markers with high sensitivity and specificity has bec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574C12N15/11
CPCC12Q1/6886C12Q2600/158G01N33/57438G01N2333/8139
Inventor 吴斌
Owner THE THIRD AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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