Application of substance for detecting FGL2 gene or FGL2 protein expression level in products for diagnosing and indicating kidney cancer

A protein expression and material technology, applied in disease diagnosis, biochemical equipment and methods, detection of programmed cell death, etc., can solve the problems of low diagnostic value of plain films, missed diagnosis of small renal cell carcinoma, misdiagnosis, and non-specificity of ultrasound examination images And other issues

Inactive Publication Date: 2017-08-04
THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

X-ray examinations usually show characteristic imaging changes in typical late-stage cases, so the diagnostic value of plain films is not great; ultrasonography images are non-specific, and it is difficult to diagnose for those with a diameter of less than 2 cm or atypical cases. Usually, experienced technicians are needed to make an accurate diagnosis, which is greatly affected by subjective factors; CT and MRI have similar diagnostic values ​​for renal cancer, and are currently the most widely used diagnostic methods for renal cancer, but a few cases cannot be qualitatively determined before surgery Diagnosis, for atypical and small renal cell carcinoma may be missed and misdiagnosed, and MRI examination is not suitable for patients with metal in the body; renal cell carcinoma usually has rich blood supply, renal arteriography may clearly show the lesion, and can be selected at the same time Renal artery embolism is conducive to the subsequent operation, but it is an invasive examination, which may increase the pain of the patient; postoperative pathological biopsy is the gold standard for diagnosis, which is conducive to diagnosis, but it has far-reaching therapeutic significance for advanced patients. no diagnostic significance

Method used

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  • Application of substance for detecting FGL2 gene or FGL2 protein expression level in products for diagnosing and indicating kidney cancer
  • Application of substance for detecting FGL2 gene or FGL2 protein expression level in products for diagnosing and indicating kidney cancer
  • Application of substance for detecting FGL2 gene or FGL2 protein expression level in products for diagnosing and indicating kidney cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Detection of FGL2 mRNA expression level in renal carcinoma tissues

[0027] 1. Materials

[0028] The cancer tissues and corresponding paracancerous tissues of 13 patients with RCC were selected. The samples were collected from patients with RCC who underwent total or partial nephrectomy (pathologically confirmed) in Chongqing Southwest Hospital from October 2015 to December 2015. Immediately after surgical resection, Renal carcinoma and paracancerous tissues were separated, numbered and stored in liquid nitrogen.

[0029] 2. Extraction of RNA from tissue samples

[0030] Use the Trizol (Takara, Cat. No. 9109) method to extract RNA in the tissue. The experimental operation is carried out according to the product instructions, and the specific operations are as follows:

[0031] (1) Put the cancer tissues and corresponding paracancerous tissue samples of 13 patients with renal cancer into a mortar that had been pre-cooled with liquid nitrogen, grind them into powder, a...

Embodiment 2

[0057] Detection of FGL2 protein expression level in fresh renal carcinoma tissue

[0058] 1. Sample

[0059] Fresh kidney cancer tissues and paracancerous tissues that were pathologically diagnosed and surgically resected in Chongqing Southwest Hospital were collected. The included cases had not received any treatment before operation, and all clinical information was collected from the patient's medical records.

[0060] 2. Western blot detection (Western blot)

[0061] The collected kidney tissue was extracted with RIPA lysate (Beyotime Biotechnology) for total protein, and the total protein concentration was determined by BCA method. Sodium dodecyl sulfate (SDS) loading buffer and protein sample were mixed at a volume ratio of 1:4, the mixed sample was placed in boiling water for 10 min to denature the protein, and cooled to room temperature. Add the protein sample to the loading well of the SDS-PAGE gel (30μg protein / well). The initial voltage of electrophoresis is 80V....

Embodiment 3

[0065] Detection of FGL2 protein expression level in paraffin-embedded renal carcinoma tissue sections

[0066] 1. Sample

[0067] Renal carcinoma tissues (paraffin-embedded, 170 cases) and paracancerous tissues (paraffin-embedded, 40 cases) diagnosed by pathology and surgically resected in Chongqing Southwest Hospital from January 2010 to December 2011 were collected. None of the included cases received any treatment before operation, and all clinical information was collected from the patient's medical records.

[0068] 2. Immunohistochemical detection

[0069] Each paraffin-embedded block was serially sectioned at 4 μm and baked at 65°C for 2 hours. Dewaxing with xylene twice (10 min / time), rehydration in graded alcohol, 0.01M citrate buffer (pH 6.0) antigen retrieval for 20 min (microwave method), hydrogen peroxide with a volume fraction of 3% at room temperature for 10 min ( Remove endogenous peroxidase), block with 5% BSA antigen for 1 h, and incubate with FGL2 mouse ...

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Abstract

The invention relates to application of a substance for detecting FGL2 gene or FGL2 protein expression level in products for diagnosing and indicating kidney cancer. According to a detection result of the substance to FGL2 gene or FGL2 protein expression level, early diagnosis on the kidney cancer or prediction on prognosis of the kidney cancer can be realized to reduce death rate of patients with the kidney cancer; treatment effect of antitumor drug can be measured by detecting the FGL2 gene or FGL2 protein expression level at a certain period after testing drug is added to cancer cells or after the testing drug is applied to tumor model animals, and effective drug capable of treating the kidney cancer can be screened. The invention further provides application of a substance for regulating FGL2 gene or lowering FGL2 protein level in preparing drug for treating the kidney cancer. The drug can treat the kidney cancer mediated by over-expression of FGL2 protein from two aspects of blocking over-expression and secretion of endogenous FGL2 protein and lowering activity of FGL2 protein.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the application of a substance for detecting the expression level of FGL2 gene or FGL2 protein in diagnosing and predicting kidney cancer products. Background technique [0002] Renal cell carcinoma is a malignant tumor that occurs in the renal parenchyma, referred to as renal cancer, which originates from the renal tubular epithelium, accounting for 3% of adult malignant tumors and 90%-95% of renal malignant tumors. It is currently the most common renal malignant tumor. The onset of RCC is hidden, and the early clinical manifestations lack specificity. Many patients have metastases when they are first diagnosed. In the past, when the typical triad of "hematuria, mass, and pain" appeared, most of them belonged to advanced tumors. Sensitive, there is no effective treatment for advanced RCC, and the prognosis is poor. Therefore, the early diagnosis of RCC is of great signif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/112C12Q2600/118C12Q2600/136C12Q2600/158G01N33/57438G01N33/57484G01N2500/00G01N2800/52G01N2800/56
Inventor 唐铭曹漵张克勤
Owner THE FIRST AFFILIATED HOSPITAL OF THIRD MILITARY MEDICAL UNIVERSITY OF PLA
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