Application of PCSK2(proprotein convertase 2) in preparation of products for detection and treatment of pancreatic neuroendocrine tumors

A neuroendocrine and pancreatic technology, applied in the field of tumor molecular biology, can solve the problems that the clinical significance of function and expression changes has not been reported, and the mechanism of action is unclear.

Inactive Publication Date: 2019-06-21
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, the mechanism of action of PCSK2 gene or protein in pancreatic neuroendocrine tumors is s

Method used

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  • Application of PCSK2(proprotein convertase 2) in preparation of products for detection and treatment of pancreatic neuroendocrine tumors
  • Application of PCSK2(proprotein convertase 2) in preparation of products for detection and treatment of pancreatic neuroendocrine tumors
  • Application of PCSK2(proprotein convertase 2) in preparation of products for detection and treatment of pancreatic neuroendocrine tumors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] This example proves that the content of PCSK2 mRNA in the samples of patients with pancreatic neuroendocrine tumors (Patients) is significantly lower than that in the samples of patients' adjacent normal pancreatic tissues (Normal).

[0032] 1. Sampling

[0033] 23 paraffin samples were obtained from the Department of Pathology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, including 7 samples of paracancerous normal pancreatic tissue (that is, normal pancreatic tissue around the tumor, which contains pancreatic neuroendocrine tissue; as a negative control) in patients with pancreatic neuroendocrine tumors , 16 patients with pancreatic neuroendocrine tumor tissue samples. The paraffin tissue was sliced, and the part exposed to the air was discarded. The thickness of the slice was about 10 μm. Every 4 wax rolls were put into a 1.5ml centrifuge tube, and 6 tubes were marked for each sample. If RNA extraction is not to be performed within a s...

Embodiment 2

[0088] Example 2 Expression of PCSK2 in pancreatic neuroendocrine tumor cell lines

[0089] Firstly, human pancreatic neuroendocrine tumor cell lines BON-1, QGP-1 and normal pancreatic epithelial cell line H6C7 were cultured, in which the cell culture medium consisted of: RPMI-1640 medium + 10% fetal bovine serum (FBS) + 1% double antibody . Culture in an incubator at 37°C, 5% CO2, and 90% relative humidity. The medium was changed once every 2 to 3 days, and 0.25% EDTA-containing trypsin was used for routine digestion and passage.

[0090] When the cell culture density reaches 80-90%, first discard the culture medium, then wash it again with PBS, suck up the residual liquid, add 1ml Trizol, blow the cells down completely and suck them into a 1.5ml centrifuge tube. If RNA extraction will not be performed within a short period of time, the samples can be stored at -80°C for a short period of time.

[0091] Extract total RNA from cultured cells as follows:

[0092] 1) Pre-coo...

Embodiment 3

[0104] Example 3 Pancreatic neuroendocrine tumor detection or prognosis assessment kit

[0105] Based on the primer set in Table 1 in Example 1, assemble the test kit for detecting pancreatic neuroendocrine tumors according to the present invention, which includes a primer pair for specific amplification of human PCSK2 gene (SEQ ID NO: 1 The forward primer shown and the reverse primer shown in SEQ ID NO:2), and the primer pair (forward primer shown in SEQ ID NO:3 and shown in SEQ ID NO:4) of specific amplification housekeeping gene (GAPDH) reverse primer); also includes SYBR Green polymerase chain reaction system, such as PCR buffer, SYBR Green fluorescent dye, dNTPs. The composition of described PCR buffer is 25mM KCl, 2.5mM MgCl 2 , 200mM (NH 4 ) 2 SO 4 ; Also includes normal pancreatic tissue cDNA: used as a negative control for quantitative PCR detection together with the cDNA of the sample to be tested, and the same amount of cDNA as the test sample is used for each r...

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Abstract

The invention discloses application of a pancreatic neuroendocrine tumor molecular marker PCSK2 (proprotein convertase 2) in the preparation of products for the detection and treatment of pancreatic neuroendocrine tumors. It is also discovered herein that expression of PCSK2 in pancreatic neuroendocrine tumor tissues or pancreatic neuroendocrine tumor cells is down-regulated. The invention also discloses a kit for detection or prognosis of pancreatic neuroendocrine tumors; the kit comprises a primer sequence for specific amplification of PCSK2. The application of the kit in the auxiliary diagnosis and prognosis estimation of pancreatic neuroendocrine tumors has guide significance to subsequent clinical treatment. In addition, pharmaceutical compositions prepared with PCSK2 to treat pancreatic neuroendocrine tumors are also clinically applicable to genetic therapy, drug therapy and other therapies of pancreatic neuroendocrine tumors.

Description

technical field [0001] The invention relates to the technical field of tumor molecular biology, in particular to the application of PCSK2 in the preparation of products for the detection and treatment of pancreatic neuroendocrine tumors. Background technique [0002] Neuroendocrine neoplasms (NEN) originate from multipotent neuroendocrine stem cells, which have neuroendocrine markers, can produce bioactive amines and / or polypeptide hormones, and have significant heterogeneity. In the past 30 years, the prevalence of NEN has increased from 1.09 / 100,000 to 5.25 / 100,000. Compared with other tumors, NEN increased more rapidly, which may be related to the advancement of diagnostic technology, more frequent tumor screening, and environmental factors. [0003] Pancreatic neuroendocrine tumors (pancreatic neuroendocrine tumors, PNET) account for about 1 / 3 of all neuroendocrine tumors, accounting for about 2% to 5% of pancreatic tumors. Although PNET is rare, it often acts on multi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886G01N33/68G01N33/574A61K45/00A61P35/00
Inventor 王文泽石秀玉肖雨
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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