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New application of polynucleotide-5'kinase-3'phosphatase

A polynucleotide and phosphatase technology, which is used in medical preparations containing active ingredients, microbial determination/inspection, instruments, etc., to achieve accurate diagnosis.

Active Publication Date: 2020-11-10
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no relevant research reports on PNKP in the diagnosis and treatment of pancreatic cancer.

Method used

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  • New application of polynucleotide-5'kinase-3'phosphatase
  • New application of polynucleotide-5'kinase-3'phosphatase
  • New application of polynucleotide-5'kinase-3'phosphatase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1, PNKP low expression in pancreatic cancer tissue

[0044] The postoperative cancer tissues and paracancerous tissues of 124 pancreatic cancer patients were collected, and the expression of PNKP protein in different tissues was analyzed by immunohistochemical staining.

[0045] (1) Experimental materials

[0046] The postoperative cancer tissues and paracancerous tissues collected from 124 pancreatic cancer patients were used to construct pancreatic cancer tissue chips; immunohistochemical kits through a commercial company.

[0047] (2) Experimental method

[0048] Before dewaxing, the tissue chips of pancreatic cancer patients should be placed in a 60°C incubator at room temperature and baked for 30 minutes. Soak in 95% ethanol for 5 minutes, soak in 85% ethanol for 5 minutes, soak in 75% ethanol for 5 minutes, rinse with tap water, and finally rinse with distilled water, 3min×2 times; heat 0.01M sodium citrate buffer solution (pH6.0) in a microwave oven Af...

Embodiment 2

[0052] Example 2, PNKP is highly expressed in normal tissues, but lowly expressed in pancreatic cancer cells

[0053] (1) Experimental materials

[0054] Human pancreatic cancer cells Patu-8988, Panc1, SW1990, CFPAC-1, ASPC-1 and BXPC-3 were preserved in our laboratory; the cell culture medium DMEM (high glucose) was produced by Hyclone Company. Trizol reagent: purchased from Invitrogen, USA; reverse transcription kit: Fermentas, USA; PCR kit: ABI company product; normal tissue cDNA was purchased from a commercial company; primers were provided by Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0055] (2) Experimental method

[0056] Human pancreatic cancer cells in the logarithmic growth phase were taken, total RNA was extracted by Trizol, and cDNA was synthesized by reverse transcription. The PCR reaction system was 25 μl, and the PCR conditions were pre-denaturation at 94°C for 3 min; denaturation at 94°C for 30 s, annealing at 60°C for 30 s, extension at 72...

Embodiment 3

[0059]Example 3. Overexpression of PNKP inhibits DNA damage of tumor cells.

[0060] (1) Experimental materials

[0061] Human pancreatic cancer cells Patu-8988 were cryopreserved in our laboratory. The cell culture medium DMEM (high sugar) is a product of Hyclone Company. comet kit: Trevigen, USA.

[0062] (2) Experimental method

[0063] The human PNKP gene was cloned into the venus vector, and venus and venus-PNKP were respectively infected into tumor cells, and were screened to become tumor cell lines with stable expression. Take the above cells in the logarithmic growth phase, inoculate them in a culture dish, and carry out UVB irradiation (2mJ / cm 2 ), to prepare cell suspension, take 30 microliters of cell suspension and mix well in 200 μL of 0.6% low-melting point agarose gel, and gently drop the mixed solution on the carrier layer covered with 1% ordinary agarose On the glass slide, after the gel is solidified, place it in the 4°C solution for 20 minutes, then ele...

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PUM

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Abstract

Provided is a new use of polynucleotide-5' kinase-3' phosphatase. In particular, provided is the use of polynucleotide-5' kinase-3' phosphatase in the preparation of products for diagnosing pancreatic cancer. Also provided is the use of triptonide as a polynucleotide-5' kinase-3' phosphatase inhibitor. Further provided is the use of polynucleotide-5' kinase-3' phosphatase as a target in the preparation of a drug for treating pancreatic cancer.

Description

technical field [0001] The present invention belongs to the field of medicine, specifically relates to the new application of polynucleotide-5'kinase-3'phosphatase, and further relates to the preparation and diagnosis of pancreatic cancer by polynucleotide-5'kinase-3'phosphatase and the application of polynucleotide-5'kinase-3'phosphatase in the preparation of drugs for treating pancreatic cancer. Background technique [0002] Pancreatic cancer is a malignant tumor derived from pancreatic cells and ductal cells. Epidemiological studies have revealed that the incidence of pancreatic cancer continues to increase. Pancreatic cancer is the malignant tumor with the worst prognosis, with a five-year survival rate of only 6%. In the past 30 years, the number of pancreatic cancer patients has increased by 6 times, and its incidence rate has increased year by year. The number of new cases of pancreatic cancer is increasing year by year. Epidemiological experts predict that in the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886G01N33/574A61K31/585A61P35/00
CPCA61K31/585A61P35/00C12Q1/68G01N33/574
Inventor 周泉生曹志飞
Owner SUZHOU UNIV
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