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Biological marker related with clear cell renal cell carcinoma and application thereof

A clear cell renal cell carcinoma and cell technology, which is applied in the field of biomarkers, can solve the problems that molecular diagnosis and targeted therapy of renal clear cell carcinoma cannot be satisfied, and the prognosis is not very ideal.

Active Publication Date: 2018-05-29
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recently, for early or localized clear cell renal cell carcinoma, surgery is the preferred treatment, including partial nephrectomy or radical resection, and the prognosis of patients after surgery is good; however, for patients with locally advanced or distant organ metastasis The prognosis of patients with clear cell renal cell carcinoma is not ideal because renal cell carcinoma is not very sensitive to radiotherapy and chemotherapy (Ljungberg B, Bensalah K, Canfield S, et al. EAU guidelines on renal cell carcinoma: 2014 update. Ehr Urol. 2015;67(5):913-924.)
[0003] In recent years, with the development of molecular biology, the potential molecular mechanism in the occurrence and development of clear cell renal cell carcinoma has become a research hotspot. Genes related to the occurrence and development of clear cell renal cell carcinoma, but the discovery of the above genes is far from meeting the needs of clinical molecular diagnosis and targeted therapy of clear cell renal cell carcinoma. To further explore the potential molecular mechanism in the development of clear cell renal cell carcinoma, Finding new biomarkers and prognostic factors is of great significance for the clinical diagnosis and treatment of clear cell renal cell carcinoma

Method used

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  • Biological marker related with clear cell renal cell carcinoma and application thereof
  • Biological marker related with clear cell renal cell carcinoma and application thereof
  • Biological marker related with clear cell renal cell carcinoma and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Example 1 Screening for Gene Markers Related to Renal Clear Cell Carcinoma

[0088] 1. Sample collection

[0089]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.

[0090] 2. Preparation of RNA samples

[0091] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0092] 3. Remove rRNA

[0093] Ribosomal RNA was removed from total RNA using the Ribo-Zero kit.

[0094] 4. Construction of cDNA library

[0095] The cDNA library was constructed using...

Embodiment 2

[0102] Example 2 QPCR sequencing to verify the differential expression of the LILRB4 gene

[0103] 1. Large-sample QPCR verification of differential expression of LILRB4 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.

[0104] 2. RNA extraction

[0105] RNA samples were extracted using the QIAGEN Tissue RNA Extraction Kit. For details, please refer to the instruction manual.

[0106] 3. Reverse transcription

[0107] mRNA reverse transcription was performed using the FastQuant cDNA First Strand Synthesis Kit (Product No.: KR106). Specific steps are as follows:

[0108] (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;

[0109] (2) Construct a 20μl reaction...

Embodiment 3

[0124] Example 3 Western blotting assay to detect the differential expression of LILRB4 protein

[0125] 1. Extraction of total tissue protein

[0126] 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.

[0127] 2. Determination of total protein concentration

[0128] The protein concentration was determined according to the instructions of the BCA protein concentration determination kit.

[0129] 3. SDS-PAGE electrophoresis

[0130] Prepare 8% separating gel and 5% stacking gel according to the instructions of the SDS-PAGE gel preparation kit and conduct electrophoresis.

[0131] 4. Western detection

[0132] 1) Electrotransfer

[0133] Put the PVDF membrane into methan...

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Abstract

The invention discloses a biological marker related with clear cell renal cell carcinoma and application thereof. The biological marker is LILRB4. The invention discloses a reagent, a kit and a chip for detecting the level of LILRB4, a medicine composition with down-regulated level of LILRB4, and application of the reagent, the kit and the chip or the medicine composition in preparation of products for diagnosing or treating the clear cell renal cell carcinoma and the invasion and transfer of the clear cell renal cell carcinoma.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a biomarker related to renal clear cell carcinoma and its application. The specific biomarker involved is LILRB4. Background technique [0002] Renal cell carcinoma, referred to as kidney cancer, originates from the parenchymal tubular epithelial system of the kidney and is one of the most common malignant tumors of the genitourinary system. The incidence rate is only lower than that of prostate cancer and bladder cancer, accounting for about 3% of all adult malignant tumors. Left and right (DeSantis CE, Lin CC, Mariotto AB, et al. Cancer treatment and survival statistics 2014. CA Cancers J Clin. 2014; 64(4):252-271.). Renal carcinoma can be divided into many different histological subtypes, among which the clear cell type is the most common, accounting for about 80-90%. Recently, for early or localized clear cell renal cell carcinoma, surgery is the preferred treatment, including parti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886G01N33/574A61K45/00A61P35/00C12Q1/02
CPCA61K45/00C12N2503/02C12Q1/6886C12Q2600/158G01N33/5011G01N33/57438G01N33/57484G01N2333/70503G01N2500/10
Inventor 宋宏涛马翠任静
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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