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A circRNA biomarker, screening method and diagnostic kit for renal clear cell carcinoma

A clear cell renal cell carcinoma, biomarker technology, applied in biochemical equipment and methods, DNA/RNA fragments, biostatistics, etc., can solve problems such as circRNA biomarkers for renal clear cell carcinoma that have not yet been found, and achieve a good consensus sexual effect

Active Publication Date: 2022-07-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] To date, no reliable circRNA biomarkers have been identified for the diagnosis of clear cell renal cell carcinoma

Method used

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  • A circRNA biomarker, screening method and diagnostic kit for renal clear cell carcinoma
  • A circRNA biomarker, screening method and diagnostic kit for renal clear cell carcinoma
  • A circRNA biomarker, screening method and diagnostic kit for renal clear cell carcinoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Analysis of circRNA expression profile in renal clear cell carcinoma

[0046] (1) Collection of serum samples from patients with renal clear cell carcinoma

[0047] After obtaining the informed consent of the patients, 6 ml of clear cell renal cell carcinoma patients and 4 healthy people with definite menstrual pathological diagnosis from April 2018 to May 2018 in the First Affiliated Hospital of Zhejiang University in Zhejiang Province were collected (as a control). . All patients with clear cell renal cell carcinoma were newly diagnosed patients and had not undergone surgery, radiotherapy and chemotherapy before blood collection. The 4 healthy controls were age-matched healthy people without malignant tumors and other diseases.

[0048] (2) Serum sample collection and processing

[0049] Draw 6ml of fasting venous blood in the morning and put it in a tube without anticoagulant, let it stand for 30 minutes, centrifuge at 3,000g for 15 minutes at 4°C, take ...

Embodiment 2

[0052] Example 2 Preliminary screening of target circRNA

[0053] (1) Collect tissue samples from patients with renal clear cell carcinoma

[0054] After obtaining informed consent from patients, 6 pairs of postoperative specimens from patients with pathologically confirmed clear cell renal cell carcinoma from April 2018 to May 2018 in the First Affiliated Hospital of Zhejiang University, Zhejiang Province were collected, including cancer tissue specimens and paired distance cancer tissue 3 All patients have not received chemotherapy and radiotherapy before surgery. All tissue samples were packed in RNase-free EP tubes and stored in a -80°C freezer.

[0055] 2. Extraction of total RNA from tissue samples

[0056] Specific steps are as follows:

[0057] (1) Weigh the tissue cut pieces with a weight between 0.3-0.5g, add 1ml of AG RNAex Pro Reagent (Aikeray Bioengineering Co., Ltd.) lysate, and add magnetic beads, use a tissue disruptor to disrupt the tissue, and homogenize t...

Embodiment 3

[0087] Example 3 Clinical small sample verification of target circRNA

[0088] 1. Collection of clinical samples

[0089] The present invention selects 7 pairs of renal clear cell carcinoma and adjacent tissue samples, they are respectively from 7 renal clear cell carcinoma patients, selects the serum of 6 healthy controls and 8 renal clear cell carcinoma patients, and the renal clear cell carcinoma tissue samples. The collection and preservation of samples were the same as those in Example 2, and the collection and preservation of serum samples were the same as those in Example 1.

[0090] 2. Extraction of total RNA

[0091] The extraction of total RNA from renal clear cell carcinoma tissue samples was the same as that in Example 2.

[0092] The extraction steps of total RNA from serum samples are as follows:

[0093] (1) Prepare 500 μl of thawed serum samples, add 2500 μl of AG RNAex Pro Reagent lysis reagent, mix well on a vortex shaker or invert several times, and let s...

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Abstract

The invention discloses a circRNA biomarker, a screening method and a diagnostic kit for renal clear cell carcinoma. The serum circRNA biomarkers of renal clear cell carcinoma provided by the present invention have the same expression in renal clear cell carcinoma tissue and serum, and the expression is abnormally significantly reduced, which is a very reliable marker for serum samples to diagnose renal clear cell carcinoma, and is a clinical The diagnosis of renal clear cell carcinoma provides a new approach. The kit provided by the invention makes the clinical diagnosis of renal clear cell carcinoma simple and fast, and the diagnosis result is reliable.

Description

technical field [0001] The invention specifically relates to a circRNA biomarker, a screening method and a diagnostic kit for renal clear cell carcinoma. Background technique [0002] Renal cell carcinoma (renal cell carcinoma for short) is one of the most common malignant tumors of the urinary system. The most common histological subtype of kidney cancer is renal clear cell carcinoma, accounting for approximately 80-90%. Currently, partial and radical nephrectomy is considered the mainstay of treatment for clear cell renal cell carcinoma. Although screening, diagnosis, and treatment of clear cell renal cell carcinoma have improved, patients with clear cell renal cell carcinoma have a high rate of local or distant metastasis and a poor prognosis, with a 5-year survival rate of less than 10%. It is worth noting that once the recurrence and metastasis of cancer in patients with clear cell renal cell carcinoma occurs, it means that the prognosis will be very unsatisfactory, r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6809C12N15/113C12N15/11G16B40/00
CPCC12Q1/6886C12Q1/6809G16B40/00C12Q2600/158C12Q2600/178C12Q2531/113C12Q2563/107
Inventor 林伟强王颖王平张云敬苏心婉
Owner ZHEJIANG UNIV
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