Renal clear cell carcinoma circRNA biomarker, screening method and diagnostic kit

A technology of clear cell renal cell carcinoma and biomarkers, applied in biochemical equipment and methods, DNA/RNA fragments, biostatistics, etc., can solve problems such as circRNA biomarkers of clear cell renal cell carcinoma that have not been discovered, and achieve good agreement sexual effect

Active Publication Date: 2021-06-18
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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  • Renal clear cell carcinoma circRNA biomarker, screening method and diagnostic kit
  • Renal clear cell carcinoma circRNA biomarker, screening method and diagnostic kit
  • Renal clear cell carcinoma circRNA biomarker, screening method and diagnostic kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0046] (1) Serum samples collected from patients with clear cell renal cell carcinoma

[0047] After obtaining the informed consent of the patients, 6 ml of early morning fasting venous blood was collected from 6 patients with clear cell renal cell carcinoma and 4 healthy subjects with clear pathological diagnosis in the First Affiliated Hospital of Zhejiang University in Zhejiang Province from April 2018 to May 2018 (as a control) . All patients with clear cell renal cell carcinoma were diagnosed for the first time, and had not undergone surgery, radiotherapy or chemotherapy before blood collection. The 4 healthy control groups were age-matched healthy people without malignant tumors or 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, centr...

Embodiment 2

[0052] Example 2 Preliminary screening of target circRNA

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

[0054] After obtaining the patient’s informed consent, 6 pairs of postoperative specimens were collected from the First Affiliated Hospital of Zhejiang University in Zhejiang Province from April 2018 to May 2018, which were confirmed by pathology as clear cell renal cell carcinoma, including cancer tissue samples and paired distance cancer tissues 3 None of the patients had received chemotherapy and radiotherapy before surgery. All tissue samples were packed into RNase-free EP tubes and stored in a -80°C refrigerator.

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

[0056] Specific steps are as follows:

[0057] ⑴Weigh the cut tissue with a weight between 0.3-0.5g, add 1ml AG RNAex Pro Reagent (Aikerui Bioengineering Co., Ltd.) lysate, and add magnetic beads, use a tissue disruptor to break the tissue, and homogeniz...

Embodiment 3

[0088] Example 3 Clinical small sample verification of target circRNA

[0089] 1. Collection of clinical samples

[0090] The present invention selects 7 pairs of clear cell renal cell carcinoma and paracancerous tissue samples, they come from 7 clear cell renal cell carcinoma patients respectively, selects the serum of 6 cases of healthy controls and 8 cases of clear cell renal cell carcinoma, and the samples of clear cell renal cell carcinoma tissue The collection and storage of samples are the same as in Example 2, and the collection and storage of serum samples are the same as in Example 1.

[0091] 2. Extraction of total RNA

[0092] The extraction of total RNA from renal clear cell carcinoma tissue samples was the same as in Example 2.

[0093] The extraction steps of total RNA from serum samples are as follows:

[0094] (1) Prepare 500 μl of thawed serum sample, add 2500 μl of AG RNAex Pro Reagent lysis reagent, mix well in a vortex shaker or upside down several time...

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Abstract

The invention discloses a renal clear cell carcinoma circRNA biomarker, a screening method and a diagnostic kit. The renal clear cell carcinoma serum circRNA biomarker provided by the invention is consistent in expression in renal clear cell carcinoma tissues and serum, and the expression is abnormally and remarkably reduced, so that the renal clear cell carcinoma serum circRNA biomarker is a very reliable marker for diagnosing the renal clear cell carcinoma in a serum sample, and a new way is provided for clinical diagnosis of the renal clear cell carcinoma. According to the kit provided by the invention, the clinical diagnosis of the renal clear cell carcinoma is simple, convenient and rapid, and the diagnosis result is reliable.

Description

technical field [0001] The present invention specifically relates to a circRNA biomarker, screening method and diagnostic kit for renal clear cell carcinoma. Background technique [0002] Renal cell carcinoma (referred to as renal cell carcinoma) is one of the most common malignant tumors of the urinary system. The most common histologic subtype of RCC is renal clear cell carcinoma, accounting for approximately 80-90%. Currently, partial and radical nephrectomy are considered the mainstay of treatment for clear cell renal cell carcinoma. Although the 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 cancer recurrence and metastasis occur in patients with clear cell renal cell carcinoma, it means that the prognosis will be very unsatisfactory, and the...

Claims

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Application Information

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Patent Type & Authority Applications(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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