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A biomarker associated with clear cell renal cell carcinoma

A technology of renal clear cell carcinoma and cells, applied in the field of biomedicine, can solve the problems of distant metastasis, fewer patients, and patient recurrence.

Active Publication Date: 2020-11-17
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The 5-year survival rate is 65%-93% for stage I and 47%-77% for stage II, but 20%-40% of patients still have recurrence or distant metastasis after surgery
Therefore, molecularly targeted drugs targeting this signaling pathway have become research hotspots and have come out successively: bevacizumab, sunitinib, sorafenib, temsirolimus , Everolimus (everolimus), pazopanib (pazopanib), and axitinib (axitinib) have become the first- and second-line drugs for the treatment of advanced metastatic renal cancer. Although they have brought encouraging results in treatment, but Few patients achieve complete remission and survive disease-free

Method used

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  • A biomarker associated with clear cell renal cell carcinoma
  • A biomarker associated with clear cell renal cell carcinoma
  • A biomarker associated with clear cell renal cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Example 1 Screening for Gene Markers Related to Renal Clear Cell Carcinoma

[0086] 1. Sample collection

[0087] Tissues from 6 cases of clear cell renal cell carcinoma and corresponding paracancerous tissues were collected. All patients had not received radiotherapy and chemotherapy before operation, and pathological examination was performed to confirm the diagnosis after operation. The informed consent of the patients was obtained for the acquisition of tissue samples, and the consent of the organizational ethics committee was obtained.

[0088] 2. Preparation of RNA samples

[0089] 1) Add liquid nitrogen to grind the tissue to powder, add 1ml TRIzol (Invitrogen) solution, mix by pipetting to fully lyse the tissue, and let stand for 5 minutes;

[0090] 2) Centrifuge at 12000rpm at 4°C for 5min, and transfer the supernatant to a 1.5ml RNase free EP tube;

[0091] 3) Add 200 μl of chloroform, vigorously shake and mix for 30 s, make the aqueous phase and organic ph...

Embodiment 2

[0108] Example 2 QPCR sequencing to verify the differential expression of SLC17A9 gene

[0109] 1. Large-sample QPCR verification of differential expression of SLC17A9 gene. According to the sample collection method in Example 1, 50 cases of paracancerous tissues and 50 cases of renal clear cell carcinoma tissues were selected.

[0110] 2. The specific steps of RNA extraction are as described in Example 1.

[0111] 3. Reverse transcription

[0112] 3. Reverse transcription: FastQuant cDNA First Strand Synthesis Kit (Product No.: KR106) was used for mRNA reverse transcription. Specific steps are as follows:

[0113] (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;

[0114] (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 μl was mixed and added to the mixture in (1) and mixed evenly;

[0115] (...

Embodiment 3

[0123] Example 3 Detection of differential expression of SLC17A9 protein by Western blotting

[0124] 1. Extraction of total tissue protein

[0125] 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.

[0126] 2. Determination of total protein concentration

[0127] The protein concentration was determined according to the instructions of the BCA protein concentration determination kit.

[0128] 3. SDS-PAGE electrophoresis

[0129] Prepare 8% separating gel and 5% stacking gel according to the instructions of the SDS-PAGE gel preparation kit and conduct electrophoresis.

[0130] 4. Western detection

[0131] 1) Electrotransfer

[0132] Put the PVDF membrane into methanol ...

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Abstract

The invention discloses a biomarker relating to renal clear cell carcinoma. The biomarker is SLC17A9. Experiments prove that the expression of the SLC17A9 is up-regulated in patients suffering from the renal clear cell carcinoma, proliferation and invasion of the renal clear cell carcinoma cells can be inhibited, and the SLC17A9 can serve as a diagnosis and / or treatment target to be applied to clinical renal clear cell carcinoma diagnosis and treatment.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a biomarker related to renal clear cell carcinoma, specifically the biomarker is SLC17A9. Background technique [0002] Renal cancer is one of the most common malignant tumors of the urinary system, and its incidence rate is second only to bladder cancer. At present, among the new cases of tumors in the world, the incidence of renal cancer is increasing at an average annual rate of 2.5%, accounting for 2%-3% of all new tumors. Kidney cancer originates from renal tubular epithelial cells in the renal parenchyma, also known as renal cell carcinoma. The pathological types of renal cell carcinoma are various, and the most common pathological type is clear cell renal cell carcinoma, accounting for about 80% to 85% of renal cell carcinomas. In recent years, the development of imaging has brought good news to patients, making most patients with RCC diagnosed as localized RCC. Surgical resectio...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K45/00A61K48/00C12Q1/6886A61P35/00A61K31/7105
CPCA61K31/7105A61K45/00A61K48/0066C12Q1/6886C12Q2600/136C12Q2600/158
Inventor 宋宏涛马翠任静
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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