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MiRNA biomarker and detection kit for thyroid cancer diagnosis

A technology for biomarkers and thyroid cancer, applied in the field of microRNA biomarkers and detection kits

Inactive Publication Date: 2016-09-21
崔学俊
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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 thyroid cancer, and there is no reliable microRNA biomarker for the diagnosis of human thyroid cancer

Method used

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  • MiRNA biomarker and detection kit for thyroid cancer diagnosis
  • MiRNA biomarker and detection kit for thyroid cancer diagnosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

[0016] 1 Objects and Methods

[0017] 1.1 Specimen source

[0018] After obtaining the patient’s informed consent, 8 pairs of postoperative specimens of patients with thyroid 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 Mini spin (USA) Eppendorfcentrifuge 5810R (USA)

[0021] Nucleic acid concentrator: Eppendorfconcentrator 5301 (USA)

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

[0023] Ultraviolet cross-linking instrument: GS GENE LINKER UV Chamber (product of BIO-RAD company)

[0024] Water bath (product of...

Embodiment 2

[0092] Example 2: Serological Analysis of Differential miRNAs in Thyroid 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 thyroid cancer and 120 healthy people in Jiangsu Provincial People's Hospital as a control. All patients with thyroid 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] Take 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-free EP t...

Embodiment 3

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

[0147] A ROC curve was constructed to compare the diagnostic ability of five serum miRNAs to distinguish thyroid cancer patients from healthy controls. The areas under the ROC curve (AUC) of the five miRNAs were: hsa-miR-193b-3p, 0.802 (95% confidence interval: 0.760-0.920); hsa-miR-384, 0.809 (95% confidence interval: 0.736-0.902); hsa-miR-145-5p, 0.804 (95% confidence interval: 0.734-0.894); hsa-miR-424-5p, 0.803 (95% confidence interval: 0.765-0.920); hsa-miR-143-3p, 0.734 ( 95% confidence interval: 0.644-0.832). Under the optimal cut off value, the sensitivity and specificity of miRNAs are as follows: hsa-miR-193b-3p, 83.3% and 82.7%, respectively; hsa-miR-384, 64.8% and 94.2%, respectively; hsa-miR -145-5p, respectively 90.7% and 61.5%; hsa-miR-424-5p, 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 0.985, and the s...

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Abstract

The invention discloses an miRNA biomarker and a detection kit for thyroid cancer diagnosis. The miRNA biomarker comprises microRNAs, namely hsa-miR-193b-3p, hsa-miR-384, hsa-miR-145-5p, hsa-miR-424-5p and hsa-miR-143-3p. The five miRNAs, namely the hsa-miR-193b-3p, the hsa-miR-384, the hsa-miR-424-5p, the hsa-miR-145-5p and the hsa-miR-143-3p, in which expression differences between the thyroid cancer and para-carcinoma tissues are obvious (difference expression amount is larger than 2 folds, and the CT value in RT-PCR is smaller than 30), are subjected to serologic expression analysis; results show that the five miRNAs are expressed stably in serum, expression and organization of serum miRNAs have high uniformity, expression of the hsa-miR-193b-3p, the hsa-miR-384 and the hsa-miR-145-5p is regulated down, and expression of the hsa-miR-424-5p and the hsa-miR-143-3p is regulated up. The five microRNAs can serve as the miRNA biomarker for thyroid cancer diagnosis, and sensitivity and specificity of combined diagnosis are higher than those of single miRNA diagnosis remarkably.

Description

technical field [0001] The invention relates to biological detection, and relates to a microRNA biomarker and a detection kit for diagnosing human thyroid cancer. Background technique [0002] Thyroid cancer is a common endocrine gland malignancy, accounting for 2.59% of the cancer incidence, including papillary thyroid carcinoma (PTC), follicular thyroid carcinoma (FTC), anaplastic thyroid carcinoma (ATC) and medullary thyroid carcinoma (MTC) , the former three originate from follicular epithelial cells, and the latter originate from parafollicular cells, of which PTC accounts for about 80% of thyroid cancers. At present, the pathogenesis of thyroid cancer is not yet clear. Research on the etiology and development mechanism of thyroid cancer has important clinical significance for improving its diagnosis rate, guiding treatment and improving prognosis. [0003] MicroRNA (miRNA) is a newly discovered non-coding small molecule RNA with a length of about 18-25 nucleotides. It...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/113C12N15/11
CPCC12Q1/6886C12Q2600/158C12Q2600/178
Inventor 崔学俊
Owner 崔学俊
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