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Application of CGREF1 as marker in diagnosis and treatment of renal clear cell carcinoma

A technology of clear cell renal cell carcinoma and drugs, which is applied in the field of biomedicine, can solve the problems that cannot meet the clinical diagnosis and treatment of clear cell renal cell carcinoma, and achieve the effect of improving the accuracy rate

Pending Publication Date: 2018-06-29
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The development of science and technology has changed the way of diagnosis and treatment of diseases. With the proposal of precision medicine, the relationship between genes and diseases has been paid more and more attention. CN201710775687.X) found genes related to the occurrence and development of clear cell renal cell carcinoma, but the genes found so far still cannot meet the needs of clinical diagnosis and treatment of clear cell renal cell carcinoma. In order to improve the diagnosis rate and survival rate of clear cell renal cell carcinoma, search for It is of great significance to predict the biological markers of renal cancer and further study the molecular mechanism of renal clear cell carcinoma

Method used

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  • Application of CGREF1 as marker in diagnosis and treatment of renal clear cell carcinoma
  • Application of CGREF1 as marker in diagnosis and treatment of renal clear cell carcinoma
  • Application of CGREF1 as marker in diagnosis and treatment of renal clear cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Example 1 Screening for Gene Markers Related to Renal Clear Cell Carcinoma

[0078] 1. Sample collection

[0079] Tissues from 6 cases of clear cell renal cell carcinoma and corresponding paracancerous tissues, and renal tissues from 4 patients with non-clear cell renal cell carcinoma were collected. All patients had not received radiotherapy and chemotherapy before operation, and postoperative pathological examination was performed to confirm the diagnosis. The informed consent of the patients was obtained for the acquisition of tissue samples, and the consent of the organizational ethics committee was obtained.

[0080] 2. Preparation of RNA samples

[0081] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0082] 3. Remove rRNA

[0083] Ribosomal RNA was removed from total RNA using the Ribo-Zero kit.

[0084] 4. Construction of cDNA library

[0085] The cDNA library was constructed usin...

Embodiment 2

[0092] Example 2 QPCR sequencing to verify the differential expression of CGREF1 gene

[0093] 1. Large-sample QPCR verification of differential expression of CGREF1 gene. According to the sample collection method in Example 1, 50 cases of paracancerous tissues and clear cell carcinoma tissues of renal clear cell carcinoma patients were selected, and 18 cases of renal tissues of non-renal clear cell carcinoma patients were collected.

[0094] 2. The specific steps of RNA extraction are as described in Example 1.

[0095] 3. Reverse transcription: FastQuant cDNA First Strand Synthesis Kit (Product No.: KR106) was used for mRNA reverse transcription. Specific steps are as follows:

[0096] (1) Add 5×gDNA Buffer 2.0μl, total RNA 1μg, add RNase Free ddH 2 O to bring the total volume to 10 μl, heat in a water bath at 42°C for 3 minutes;

[0097] (2) Construct a 20μl reaction system, 10× Fast RT Buffer 2.0μL, RT Enzyme Mix 1.0μl, FQ-RT Primer Mix 2.0μl, RNase Free ddH 2 O 5.0 μ...

Embodiment 3

[0112] Example 3 Western blot assay to detect differential expression of CGREF1 protein

[0113] 1. Extraction of total tissue protein

[0114] Cut the tissue with scissors and put it into a glass homogenizer placed in ice. Mix the RIPA lysate and PMSF at a ratio of 100:1, and add the corresponding amount of RIPA lysate, crush the tissue with a glass homogenizer until it is fully lysed, suck the lysed liquid into an EP tube, centrifuge at 14,000 rpm for 5 min at 4°C, and collect the supernatant.

[0115] 2. Determination of total protein concentration

[0116] The protein concentration was determined according to the instructions of the BCA protein concentration determination kit.

[0117] 3. SDS-PAGE electrophoresis

[0118] Prepare 8% separating gel and 5% stacking gel according to the instructions of the SDS-PAGE gel preparation kit and perform electrophoresis.

[0119] 4. Western detection

[0120] 1) Electrotransfer

[0121] Put the PVDF membrane into methanol solut...

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Abstract

The invention discloses application of CGREF1 as a marker in diagnosis and treatment of renal clear cell carcinoma. The invention discloses a product including a reagent for detecting the expression level of the CGREF1 and application of relevant reagents in preparing the product for diagnosing the renal clear cell carcinoma; meanwhile, the invention discloses a pharmaceutical composition including an inhibitor for inhibiting the expression level of the CGREF1 and application of relevant inhibitors in preparing the pharmaceutical composition for treating the renal clear cell carcinoma; the invention also discloses application of the CGREF1 in screening candidate compounds for treating the renal clear cell carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the application of CGREF1 as a biomarker in the diagnosis and treatment of renal clear cell carcinoma. Background technique [0002] Renal cell carcinoma (Renal Cell Carcinoma, RCC) accounts for 2%-3% of all human malignant tumors, among which clear cell renal cell carcinoma (ccRCC) is the most common, accounting for 80% to 90% of the total (Siegel R. Naishadham D. Jemal A. Cancer statistics, 2012. CA: a cancer journal forclinicians 2012.62(1):10-29.). Renal cell carcinoma is not sensitive to radiotherapy and chemotherapy. Once advanced renal cell carcinoma metastasizes, the prognosis is extremely poor. In clinical practice, in addition to surgical resection of the primary tumor of renal cell carcinoma, tumor reduction surgery is currently clinically effective for metastatic renal cell carcinoma. Systemic targeted therapy for cell carcinoma, including TKI and mTOR inhibitors, as well as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/574
CPCC12Q1/6886C12Q2600/118C12Q2600/158C12Q2600/178G01N33/57407
Inventor 任静马翠
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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