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Diagnosis marker and treatment target of renal clear cell carcinoma

A renal clear cell carcinoma and substance technology, applied in the field of biomedicine, can solve the problems of unsatisfactory treatment effect and lack of treatment methods for renal cancer patients.

Inactive Publication Date: 2017-11-24
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because renal cancer is not sensitive to radiotherapy and chemotherapy, there is still a lack of very effective treatment methods for patients with advanced renal cancer and postoperative recurrence and metastasis of renal cancer, although the development of molecular targeted drug therapy has improved the prognosis of such patients , but the overall treatment effect of renal cancer patients is still unsatisfactory

Method used

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  • Diagnosis marker and treatment target of renal clear cell carcinoma
  • Diagnosis marker and treatment target of renal clear cell carcinoma
  • Diagnosis marker and treatment target of renal clear cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1 Screening for Gene Markers Related to Renal Clear Cell Carcinoma

[0066] 1. Sample collection

[0067]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.

[0068] 2. Preparation of RNA samples

[0069] 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;

[0070] 2) Centrifuge at 12000rpm at 4°C for 5min, and transfer the supernatant to a 1.5ml RNase free EP tube;

[0071] 3) Add 200 μl of chloroform, vigorously shake and mix for 30 s, make the aqueous phase and organic pha...

Embodiment 2

[0088] Example 2 QPCR sequencing to verify the differential expression of MRO gene

[0089] 1. Large-sample QPCR verification of differential expression of MRO genes. 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.

[0090] 2. RNA extraction The specific steps are as described in Example 1.

[0091] 3. Reverse transcription

[0092] 3. Reverse transcription: FastQuant cDNA First Strand Synthesis Kit (Product No.: KR106) was used for mRNA reverse transcription. Specific steps are as follows:

[0093] (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;

[0094] (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;

[0095] (3) Heat in...

Embodiment 3

[0109] Example 3 Western blot assay to detect differential expression of MRO protein

[0110] 1. Extraction of total tissue protein

[0111] 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.

[0112] 2. Determination of total protein concentration

[0113] The protein concentration was determined according to the instructions of the BCA protein concentration determination kit.

[0114] 3. SDS-PAGE electrophoresis

[0115] Prepare 8% separating gel and 5% stacking gel according to the instructions of the SDS-PAGE gel preparation kit and perform electrophoresis.

[0116] 4. Western detection

[0117] 1) Electrotransfer

[0118] Put the PVDF membrane into methanol solution...

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Abstract

The invention discloses a diagnosis marker and treatment target of renal clear cell carcinoma and belongs to the technical field of gene new application. The diagnosis marker and treatment target is MRO. The diagnosis marker and treatment target has the advantages that the fact that the expression of the marker MRO in a renal clear cell carcinoma is reduced is discovered for the first time, the proliferation, migration and invasion of cancer cells can be lowered by increasing the expression of the MRO, and the MRO serving as the target can be used for treating the renal clear cell carcinoma and renal carcinoma metastasis.

Description

technical field [0001] The invention belongs to the field of biomedicine, and relates to a diagnostic marker and a therapeutic target of renal clear cell carcinoma, specifically, the diagnostic marker and a therapeutic target are MRO. Background technique [0002] Renal cell carcinoma (RCC), also known as renal adenocarcinoma, referred to as kidney cancer for short, is a malignant tumor originating from renal tubular epithelial cells, and its incidence rate ranks second among urological tumors. In recent years, the incidence of kidney cancer has shown a clear upward trend. Clear cell renal cell carcinoma (ccRCC) is the most common histological type of renal cancer, accounting for about 80-85%. Radical nephrectomy is currently the most important method for the treatment of RCC. Surgical resection has the most definite curative effect on RCC whose mass is confined to the perirenal fascia, but has very limited curative effect on patients who have already infiltrated and metast...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/118C12Q2600/158G01N33/57438G01N33/68G01N2500/00
Inventor 任静马翠石小峰
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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