miRNA biomarker and detection kit used for urethral carcinoma diagnosis

A technology for biomarkers and urethral cancer, which is applied in the determination/testing of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., and can solve the problems of no reliable microRNA biomarkers.

Inactive Publication Date: 2016-07-13
青海七彩花生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are few studies on microRNA biomarkers for the diagnosis of human urethr

Method used

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  • miRNA biomarker and detection kit used for urethral carcinoma diagnosis
  • miRNA biomarker and detection kit used for urethral carcinoma diagnosis
  • miRNA biomarker and detection kit used for urethral carcinoma diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1: Screening of differential miRNAs in urethral cancer tissues

[0016] 1 Objects and Methods

[0017] 1.1 Specimen source

[0018] After obtaining the informed consent of the patients, 8 pairs of postoperative specimens of patients with urethral cancer confirmed by pathology in Jiangsu Provincial People's Hospital from May 2013 to September 2014 were collected, including cancer tissue specimens and paired paracancerous tissues that were more than 3 cm away from the cancer tissue All patients had not received chemotherapy and radiotherapy before operation.

[0019] 1.2 Main instruments and equipment

[0020] Desktop centrifuge: Eppendorf Minispin (USA) Eppendorfcentrifuge5810R (USA)

[0021] Nucleic acid concentrator: Eppendorfconcentrator5301 (USA)

[0022] Ultraviolet spectrophotometer: DU640, balance (Beckman company product)

[0023] Ultraviolet crosslinking instrument: GSGENELINKERUVChamber (product of BIO-RAD company)

[0024] Water bath (product of ...

Embodiment 2

[0092] Example 2: Serological Analysis of Differential miRNAs in Urethral Cancer Tissues

[0093] 1 Objects and Methods

[0094] 1.1 Specimen source

[0095] After obtaining the informed consent of the patients, 6ml of fasting venous blood in the morning from May 2013 to July 2014 was collected from 165 patients with pathologically diagnosed urethral cancer and 120 healthy people in Jiangsu Provincial People's Hospital as a control. All patients with urethral cancer were diagnosed for the first time, and had not undergone surgery, radiotherapy or chemotherapy before blood collection. The 120 healthy control groups were age-matched healthy people without malignant tumors or other diseases.

[0096] 1.2 Serum sample collection and processing

[0097] 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 1300g (or 4000rpm) at 4°C for 15 minutes, take the upper serum and divide it into RNase-freeEP...

Embodiment 3

[0146] Embodiment 3: receiver operating characteristic curve (ROC) analysis

[0147] A ROC curve was constructed to compare the diagnostic ability of 5 serum miRNAs in distinguishing urethral cancer patients from healthy controls. The areas under the ROC curve (AUC) of the five miRNAs were: hsa-miR-384, 0.814 (95% confidence interval: 0.760-0.920); hsa-miR-155-5p, 0.812 (95% confidence interval: 0.736-0.902); hsa-miR-424-5p, 0.818 (95% confidence interval: 0.734-0.894); hsa-miR-132-3p, 0.805 (95% confidence interval: 0.765-0.920); hsa-miR-143-3p, 0.743 ( 95% confidence interval: 0.644-0.832). Under the optimal cutoff value, the sensitivity and specificity of miRNAs are as follows: hsa-miR-384, 83.3% and 82.7%, respectively; hsa-miR-155-5p, 64.8% and 94.2%, respectively; hsa-miR- 424-5p, respectively, 90.7% and 61.5%; hsa-miR-132-3p, respectively, 70.4% and 82.5%; hsa-miR-143-3p, respectively, 67.3% and 74.1%. The combined AUC of these five miRNAs can reach 0.983, and the se...

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Abstract

The invention discloses a miRNA biomarker and a detection kit used for urethral carcinoma diagnosis. The miRNA biomarker consists of the following microRNAs: hsa-miR-384, hsa-miR-155-5p, hsa-miR-424-5p, hsa-miR-132-3p and hsa-miR-143-3p. According to the miRNA biomarker and the detection kit, the five microRNAs, including the hsa-miR-384, the hsa-miR-155-5p, the hsa-miR-424-5p, the hsa-miR-132-3p and the hsa-miR-143-3p, remarkable in urethral carcinoma and para-carcinoma tissue expression difference are subjected to serological expression analysis; and a result shows that the five miRNAs are stably expressed in serum, the expression of the miRNAs in the serum has very good consistency with tissues, the expression of the hsa-miR-384, the hsa-miR-155-5p and the hsa-miR-424-5p is down-regulated, and the expression of the hsa-miR-132-3p and the hsa-miR-143-3p is up-regulated. The five miRNAs can serve as biomarkers for the urethral carcinoma diagnosis, and the sensitivity and specificity of combined diagnosis are remarkably higher than those of single miRNA diagnosis.

Description

technical field [0001] The invention relates to biological detection, and relates to a microRNA biomarker and a detection kit for diagnosing human urethral cancer. Background technique [0002] Urethral cancer is a urothelial tumor that can occur in both sexes. It is relatively rare clinically, and most of them are males. It is the most common tumor in the urethral tumor and is a malignant tumor. According to its origin, it is divided into squamous epithelial carcinoma, transitional epithelial carcinoma, adenocarcinoma, clear cell carcinoma, urethral cloacal carcinoma, adenoid cystic carcinoma, carcinoid, mixed carcinoma, cowper urethral accessory adenocarcinoma. Squamous cell carcinoma is the most common, mostly occurring in the anterior or middle urethra, transitional epithelial carcinoma mostly occurs in the posterior urethra, and adenocarcinoma mostly occurs in the middle urethra. Urethral cancer is mainly treated with surgery. Radiation therapy can be used as preopera...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 陈亮
Owner 青海七彩花生物科技有限公司
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