Application of FBXO31 gene and related products in preparing gastric cancer diagnostic reagent

A diagnostic reagent and gene technology, applied in the field of bioengineering, can solve problems such as unclear expression level and function of FBXO31

Inactive Publication Date: 2014-09-03
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is unclear what the expression level and function of FBXO31

Method used

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  • Application of FBXO31 gene and related products in preparing gastric cancer diagnostic reagent
  • Application of FBXO31 gene and related products in preparing gastric cancer diagnostic reagent
  • Application of FBXO31 gene and related products in preparing gastric cancer diagnostic reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] RT-PCR, qRT-PCR and Western blot showed that the mRNA and protein levels of FBXO31 were significantly decreased in human gastric cancer tissues;

[0061] 1. Materials and methods

[0062] (1) Patient

[0063] 56 cases of gastric cancer and their surrounding tissues were obtained from patients who underwent gastric tumor resection in Shandong Cancer Hospital during 2012-2013, and the specimens were stored at -80°C. The age, gender, pathological type, degree of differentiation, tumor size (diameter), invasion, local and distant metastasis of the patient at the time of surgery were statistically analyzed. The details are shown in Table 1:

[0064] Table 1. Patient and tumor characteristics and the expression level of FBXO31 protein in normal mucosa and gastric cancer tissues

[0065]

[0066]

[0067] 1. Pathological tumor-node-metastasis.

[0068] 2. Histological differentiation status: LDA, poorly differentiated adenocarcinoma; MDA, moderately differentiated a...

Embodiment 2

[0098] The relationship between the expression of FBXO31 detected by immunohistochemistry and the clinical parameters and postoperative survival of patients with gastric cancer

[0099] 1. Materials and methods

[0100] (1) Tissue chip

[0101] Paraffin-embedded human gastric cancer tissue chips were purchased from Shanghai Xinchao Co., Ltd. One chip contains 180 spots, including 90 specimens of gastric cancer patients. Each patient contains two points, one point represents the cancer tissue, and the other point represents the corresponding paracancerous tissue. Information about the patient's sex, age, tumor size, clinical data, and survival period were all provided by the company.

[0102] (2) Immunohistochemical detection

[0103] ① Dewaxing and hydration: Bake tissue sections at 60°C for 2 hours before dewaxing

[0104] a. Xylene dewaxing 5mim×3 times

[0105] b. Soak in absolute ethanol for 5min×2 times

[0106] c. Soak in 95% ethanol for 5min×2 times

[0107] d. ...

Embodiment 3

[0133] Effect of FBXO31 on the proliferation ability and cell cycle progression of gastric cancer cells detected by colony formation assay and flow cytometry

[0134] 1. Materials and methods

[0135] (1) Experimental materials and cell culture

[0136] Human gastric cancer cell lines BGC-823, HGC-27 were purchased from Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences.

[0137] Cell culture at 37°C, 95% air, 5% CO 2 constant temperature incubator.

[0138] The medium was 1640 medium (purchased from Invitrogen, Carlsbad, CA) containing 10% fetal bovine serum (FBS), 100 U / mL penicillin and 2 mmol / L L-glutamine.

[0139] Human miR-20a mimics, miR-17 mimics, blank control mimics, miR-20a inhibitors, miR-17 inhibitors, and blank control inhibitors were purchased from Ribobio (Guangzhou, China).

[0140] Both the pCMV-myc-FBXO31 vector and the F-box sequence deletion mutation vector pCMV-myc-FBXO31ΔF were constructed according to the methods in th...

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Abstract

The invention relates to application of an FBXO31 gene and related products in preparing a gastric cancer diagnostic reagent. The related products of the FBXO31 gene are one or more of spliceosome of the FBXO31 gene, antisense oligodeoxynucleotide of the FBXO31 gene, mRNA (Messenger Ribonucleic Acid) of the FBXO31 gene, a protein of the FBXO31 gene, a peptide fragment encoded by the FBXO31 gene, an antibody of the protein of the FBXO31 gene, a regulating small molecular RNA related to the FBXO31 gene or an inhibitor of the regulating small molecular RNA related to the FBXO31 gene. The invention provides a novel concept of diagnosing gastric cancer and provides a novel path for researching and developing the gastric cancer diagnostic reagent.

Description

technical field [0001] The invention relates to the application of FBXO31 gene and related products in the preparation of gastric cancer diagnostic reagents, and belongs to the technical field of bioengineering. Background technique [0002] Gastric cancer is one of the most common malignant tumors worldwide, and its mortality rate ranks second in cancer-related deaths. According to statistics in 2008, the incidence of gastric cancer accounts for 8% of the total incidence of cancer, while the mortality rate of gastric cancer accounts for 10% of cancer-related deaths. East Asia is the region with the highest incidence of gastric cancer, especially China. Because gastric cancer is difficult to diagnose in its early stages, most patients are diagnosed at an advanced or metastatic stage. With traditional surgical treatment, chemotherapy or radiotherapy do not do much to improve the survival rate of gastric cancer patients. Therefore, it is critical to find new diagnostic or p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68G01N33/574
CPCC12Q1/6886C12Q2600/158C12Q2600/178G01N33/57446G01N2333/47
Inventor 刘志方张新潮徐霞孔叶邹水燕
Owner SHANDONG UNIV
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