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Application of protocadherin (PCDH) 17 genes

A gene and methylation technology, applied in the application field of PCDH17 gene, can solve problems such as biological function and possible mechanism of action are not elucidated

Active Publication Date: 2012-08-08
THE AFFILIATED SIR RUN RUN SHAW HOSPITAL OF SCHOOL OF MEDICINE ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Based on the above research basis, we believe that PCDH17 may play the role of a tumor suppressor gene in gastrointestinal tumors, but its biological function and possible mechanism of action in gastrointestinal tumors have not yet been elucidated

Method used

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  • Application of protocadherin (PCDH) 17 genes
  • Application of protocadherin (PCDH) 17 genes
  • Application of protocadherin (PCDH) 17 genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Detection of expression level and methylation status of PCDH17.

[0028] 1. Tissue Isolation

[0029] Human gastrointestinal tissue samples were obtained from patients with primary adenocarcinoma confirmed by pathology who underwent direct surgical resection without neoadjuvant therapy from February 2004 to June 2006 at Run Run Shaw Hospital Affiliated to Zhejiang University. surgical specimens. Each surgical specimen included tumor tissue, paracancerous tissue, and corresponding normal tissue far from the tumor (more than 5 cm away from the tumor). After the gastrointestinal tissue specimens were obtained from the operating room, the specimens at each site were immediately divided into three parts, and the two parts were immediately stored in liquid nitrogen, one of which was used to extract RNA and the other was used to extract protein. The third part was fixed in formalin for 24 h, and then sent to the pathology department for paraffin embedding for later...

Embodiment 2

[0049] Example 2 Demethylation reagent 5-Aza and histone deacetylase inhibitor TSA treated cells.

[0050] The PCDH17 expression silenced cell line was inoculated in a 10 cm diameter petri dish, and 1×106 cells were inoculated in each dish. After overnight culture, the cells were treated with demethylation reagent 5-Aza (final concentration 10 μmol / L) for three days, and then treated with histone deacetylase inhibitor TSA (final concentration 300 nmol / L) for 24 hours. Finally, the above-mentioned treated cells were collected, and DNA and RNA were extracted for future use.

[0051] The experimental results showed that after the gastric cancer cell lines YCCEL1 and SNU719 with silenced PCDH17 expression and the intestinal cancer cell lines HCT116 and SW480 were treated with 5-Aza and TSA, RNA and DNA were extracted again to detect the expression level and methylation status of PCDH17, and the results found four The expression of PCDH17 was repaired to varying degrees in all cel...

Embodiment 3

[0052] Example 3 Representative BGS Results.

[0053] We detected the detailed methylation profiles of 37 CpG island sites in the PCDH17 gene by BGS, and the detected 37 CpG island sites also included the CpG island sites analyzed by MSP. The results showed that the methylation rate of PCDH17 CpG island was very low in normal gastric mucosal tissue and in the cell line HCT116 / DKO in which DNMTs were knocked out, while in gastric cancer cell lines YCCEL1, SNU719 and intestinal cancer cell lines that did not express PCDH17 Densely methylated CpG island sites in HCT116, SW480 were detected in large numbers. Moreover, the methylation status of the CpG island was detected again after treating the gastric cancer cell lines YCCEL1, SNU719 and intestinal cancer cell lines HCT116 and SW480 that did not express PCDH17 with 5-Aza and TSA, and the BGS results showed that the PCDH17 CpG island site was obviously demethylated , which is consistent with the MSP results of Example 1. The re...

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Abstract

An application of protocadherin (PCDH) 17 genes belongs to the technical field of genes. On one hand the PCHD17 genes can be used as a mark of early diagnosis of gastric cancer and colorectal cancer and is favorable for early monitoring and prevention of gastric cancer and colorectal cancer, and on the other hand the gene can be used as a new target spot for chemotherapy sensibility detection of gastric cancer and colorectal cancer and is favorable for guiding chemotherapy of gastric cancer and colorectal cancer.

Description

technical field [0001] The invention belongs to the field of gene technology, and in particular relates to an application of PCDH17 gene. Background technique [0002] Gastrointestinal tumors are the most common malignant tumors in humans. According to statistics, gastrointestinal tumors account for about 20% of all tumors in the world. The incidence of gastric cancer and colorectal cancer still ranks third to fourth in the world, and they also occupy the second to third in the death rate caused by cancer. Although comprehensive tumor treatment based on surgery, chemotherapy, and radiotherapy has significantly improved and increased the cure rate of patients with gastrointestinal tumors, there is still a lack of specific detection methods and treatments for early diagnosis and early treatment of gastrointestinal tumors. method, so it is very urgent to find a new tumor marker. The current research has found that the occurrence of gastric cancer goes through four stages: atr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 何超胡晓彤陶谦隋新兵
Owner THE AFFILIATED SIR RUN RUN SHAW HOSPITAL OF SCHOOL OF MEDICINE ZHEJIANG UNIV
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