Application of BRSK2 in preparation of reagents for diagnosing pancreatic cancer

A technique for pancreatic cancer and reagents, which is applied in the fields of medicine and genetic engineering, and can solve problems such as the abnormal formation of tubulin, the influence of axon formation, and the loose combination of MAPs and tubulin.

Inactive Publication Date: 2013-10-23
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

One possible mechanism is that SAD phosphorylates Tau, which causes MAPs to not bind tightly to tubulin, resulting in the abnormal formation of tubulin and ultimately affecting axon formation.

Method used

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  • Application of BRSK2 in preparation of reagents for diagnosing pancreatic cancer
  • Application of BRSK2 in preparation of reagents for diagnosing pancreatic cancer
  • Application of BRSK2 in preparation of reagents for diagnosing pancreatic cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1 Immunohistochemical Analysis of the Expression of BRSK2 in Pancreatic Cancer Tissue

[0044] 1) Sample processing:

[0045] Take a tissue block of moderate size, generally 2.5cm×2.5cm×0.2cm. Tissues were fixed with 4% paraformaldehyde, and the fixation time was preferably within 12 hours, and generally the fixation time should not exceed 24 hours.

[0046] 2) Tissue dehydration, transparency, wax soaking:

[0047] After fixation, the tissues were dehydrated, cleared, dipped in wax and embedded. The principle to be mastered is that the dehydration and transparency should be sufficient but not overdone, the wax immersion time should be long enough, and the temperature should not be high, otherwise the tissue will be hard and brittle and it will be difficult to slice the tissue. during dyeing process

[0048] It is easy to peel off, which is not conducive to the positioning and background of immunohistochemical staining antigens, so the time for dehydration w...

Embodiment 2

[0054] Example 2 Tissue microarray analysis of the distribution of BRSK2 in pancreatic cancer and normal pancreatic tissue

[0055] Pancreatic cancer samples from different sources and normal pancreas samples were made into tissue microarrays, and the cells were stained brownish yellow as a positive standard. The scoring system is divided into 0 to 3 points, 0 points for colorless, 1 point for yellow, 2 points for brown, and 3 points for brown. 0,1 is negative, 2,3 is positive. Each group of samples was scored 3 times, and scored separately according to positive and negative. The statistical results showed that 25 of the 27 tumor samples were BRSK2-positive (96.2%), and 2 were BRSK2-negative (3.8%); among the 23 normal exocrine pancreatic tissues, 2 were BRSK2-positive (8.7%), and 21 were BRSK2-positive (8.7%). BRSK2 negative (91.3%). Gradpad Prism software chi-square test showed that pancreatic cancer was significantly correlated with BRSK2 expression (P<0.001).

Embodiment 3

[0056] Example 3 Western blot detection of endogenous BRSK2 protein in various pancreatic cancer cells

[0057] 3.1 Collection of various pancreatic cancer cell protein samples

[0058] 1) Take out the cryotube from the liquid nitrogen tank, put it directly into warm water at 37°C, and shake it from time to time to melt it as soon as possible.

[0059] 2) Take out the cryotube from the 37°C water bath, suck out the cell suspension with a pipette, pour into the centrifuge tube and add more than 10 times of culture medium, mix and centrifuge at low speed, discard the supernatant, and wash with culture medium again.

[0060] 3) After properly diluting with the culture medium, inoculate the culture bottle, place it in a 37°C incubator for static culture, replace the culture medium the next day, and continue the culture. Subculture when cultured to a certain concentration. (PANC-1 cells were cultured in DMEM high-glucose medium containing 10% Gibco fetal bovine serum, Miapaca-2, ...

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Abstract

The invention belongs to the fields of medicine and gene engineering, and relates to new application of a neural differentiation related gene BRSK2. The invention provides new application of human BRSK2, namely application of human BRSK2 in preparation of reagents for diagnosing pancreatic cancer. Particularly, the BRSK2 can be used as an auxiliary molecular marker for diagnosing pancreatic cancer. The BRSK2 is highly expressed in pancreatic cancer tissues or cells, and is lowly or not expressed in normal tissues or para-carcinoma tissues. The pancreatic cancer risk can be judged by detecting the BRSK2 content in the sample. The invention also provides a kit and method for detecting the BRSK2 content.

Description

technical field [0001] The invention belongs to the fields of medicine and genetic engineering, and relates to a new application of a gene related to nerve polarity differentiation-BRSK2. Background technique [0002] Pancreatic cancer is a malignant tumor formed by the malignant transformation of pancreatic cancer epithelium. Although its incidence rate ranks only 13th among malignant tumors, its lethality ranks 8th among malignant tumors. According to the World Health Organization, more than 216,000 people worldwide are diagnosed with pancreatic cancer each year. In my country, the incidence of this cancer was originally very low, but it has also shown an increasing trend in recent years, and has risen to the fifth place in recent years. Even with chemotherapy, radiotherapy, surgery and other treatment methods, the survival rate of pancreatic cancer is still less than 5%. Because metastases are already present when primary symptoms appear, pancreatic cancer is often diag...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/574
Inventor 余龙王颖黎唐丽莎
Owner FUDAN UNIV
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