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Application of circulating nucleic acid serving as breast cancer biomarker

A breast cancer and nucleic acid technology, which is applied in the application field of circulating nucleic acid as a breast cancer biomarker, can solve the problems of small sample size, lack of research, poor repeatability of results, etc.

Active Publication Date: 2017-10-27
KUNSHAN RNAI INST
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AI Technical Summary

Problems solved by technology

[0009] However, although the research on breast cancer-related circulating miRNA has a certain foundation, the existing results mostly focus on the role of distinguishing breast cancer patients from healthy people, that is, the discussion of circulating miRNA as a marker for early diagnosis of breast cancer, and the role of circulating miRNA as a biomarker for early diagnosis of breast cancer. Although there is a small amount of research on breast cancer prognostic markers related to metastasis and molecular subtypes of breast cancer patients, the sample size in existing studies is small, and the results of different research groups are compared repeatedly Difference
There are few studies on the relationship between circulating miRNAs and the clinical outcome and survival of breast cancer patients, especially the studies on patients with long-term follow-up based on large samples are very scarce

Method used

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  • Application of circulating nucleic acid serving as breast cancer biomarker
  • Application of circulating nucleic acid serving as breast cancer biomarker
  • Application of circulating nucleic acid serving as breast cancer biomarker

Examples

Experimental program
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Effect test

Embodiment 196

[0141] Example 1 Determination of 96 candidate miRNAs

[0142] In order to find circulating miRNAs that can be effectively used as prognostic markers for breast cancer, this study screened multiple breast cancer-related miRNAs in reported databases or studies, but their circulating forms have not been studied yet. Using this as a candidate gene, the expression profile analysis was performed in the multi-sample training set. Sources and selection criteria of candidate genes: 1) miRNAs whose expression is different (>2-fold) in breast cancer and para-cancerous tissues, as indicated by microarray or functional experiments; 2) with prognostic indicators such as breast cancer type and lymph node status Related miRNAs; 3) MiRNAs that have been shown to participate in the development of breast cancer in cell or animal models; 4) Potentially effective miRNAs have been detected in the serum of small breast cancer populations; 5) Widely present in serum plasma miRNA. Through the above...

Embodiment 2

[0143] Example 2 Screening of internal reference and standardization of miRNA expression

[0144] 25 breast cancer patients and 20 age-matched healthy controls were randomly selected to detect the expression profiles of 96 miRNA micro-serum extraction-free multiplex qRT-PCR, and the obtained miRNA Ct values ​​were stabilized by NormFinder and geNorm software gender assessment.

[0145] NormFinder analysis showed that miR-132 was the most stable miRNA expressed in patients and healthy controls, while geNorm analysis suggested that miR-103a / miR-107 was suitable as an internal reference. The evaluation of the stability of the pairwise combination of the above three miRNAs (miR-132, miR-103a, miR-107) showed that miR-103a / miR-132 had the strongest stability and was suitable as an internal reference.

[0146] After selecting the internal control, normalization was performed on the remaining miRNAs from breast cancer patients and healthy controls. Such as figure 1 As shown, differe...

Embodiment 3

[0147] Example 3 Acquisition of circulating miRNA expression profile in breast cancer patients

[0148] In order to obtain an effective combination of miRNAs for predicting the outcome of breast cancer, the original data of 96 miRNAs from 189 breast cancer patients were screened according to the following criteria: 1) The Ct value of circulating miRNAs was between 15 and 35; The Ct value of miRNA in the repeated serum experiments was STD<2; 2) the Ct value of miRNA was between the average Ct±3 of 189 breast cancer patients. Finally, the expression profiles of 82 serum miRNAs from 182 breast cancer patients meeting the requirements were obtained. The clinical information of 182 breast cancer patients is shown in Table 1, with a median age of 52 years. Among them, 55 breast cancer patients had adverse clinical outcomes, including recurrence in situ, distant metastasis and death.

[0149] Using miR-103a / miR-132 as an internal reference, the Ct values ​​of the other 80 miRNAs wer...

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Abstract

The invention discloses application of circulating nucleic acid serving as a breast cancer biomarker. Via a large number of literature researches, 96 miRNAs which have differential expression in a breast cancer tissue and body fluid or have functional verification for a level of a breast cancer cell line are screened out and used as candidate genes for researching; particularly, a prognosis relationship between the circulating miRNAs and long-term follow-up people suffering from the breast cancer is researched in the application; for 182 patients, based on bivariate regression analysis of machine training, 2 to 10 miRNA combinations have a distinguishing effect on prognosis of the breast cancer patients; multi-factor Cox regression analysis shows that a combination of 2 to 7 miRNAs is used as an independent risk factor of disease free survival and distant disease free survival of the breast cancer patients; particularly in lymph node-negative patients, a combination of 4 to 5 miRNAs has a better survival forecasting effect on the low and high-risk breast cancer patients. The research provides possibility for the application of the circulating miRNA in serving as a tumor marker.

Description

technical field [0001] The present invention relates to the use of circulating nucleic acids as biomarkers for breast cancer. Specifically, the present invention relates to the application of circulating microRNA (miRNA) in the prognosis of breast cancer; more specifically, the present invention relates to the application of the combination of 2-10 different miRNAs in the prognosis of breast cancer, including the application based on the 2- The combination of 10 different miRNAs is used to prepare a kit for breast cancer prognosis, a microarray, and a method for predicting the prognosis of breast cancer by using the kit and the microarray. Background technique [0002] Breast cancer is the cancer with the highest incidence in women worldwide, accounting for 23% of all cancers, and breast cancer is also the leading cause of cancer death in women. According to the 2012 statistics of the World Health Organization (GLOBOCAN 2012), compared with 2008, in the past four years, the...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/118C12Q2600/166C12Q2600/178
Inventor 不公告发明人
Owner KUNSHAN RNAI INST
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