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Application of Circulating Nucleic Acids as Biomarkers for Breast Cancer

A breast cancer and nucleic acid technology, applied in the application field of circulating nucleic acid as a breast cancer biomarker, can solve the problems of lack of research, poor repeatability of results, small sample size, etc., and achieve the effect of predicting survival effect and valuable application value

Active Publication Date: 2021-07-27
KUNSHAN RNAI INST
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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 Acids as Biomarkers for Breast Cancer
  • Application of Circulating Nucleic Acids as Biomarkers for Breast Cancer
  • Application of Circulating Nucleic Acids as Biomarkers for Breast Cancer

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 effective as prognostic markers for breast cancer, this study screened multiple breast cancer-related miRNAs in reported databases or studies, while their circulating forms have not been studied. Using this as a candidate gene, expression profiling was performed in the multi-sample training set. Sources and selection criteria of candidate genes: 1) miRNAs with differential (>2-fold) expression indicated by microarray or functional experiments in breast cancer and adjacent tissues; 2) prognostic indicators such as breast cancer typing and lymph node status Relevant miRNAs; 3) miRNAs that have been proved to be involved in the occurrence and development of breast cancer in cell or animal models; 4) miRNAs that may be effective have been detected in the serum of a small sample of breast cancer populations; 5) are widely present in serum and plasma. miRNAs. Through the ...

Embodiment 2

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

[0144] Twenty-five breast cancer patients and 20 age-matched healthy controls were randomly selected for detection of 96 miRNA trace serum extraction-free multiplex qRT-PCR expression profiles, and the obtained miRNA Ct values ​​were stabilized by NormFinder and geNorm software Sexual 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 were suitable as internal controls. The stability evaluation 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. like figure 1 As shown, different m...

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 present invention is the application of circulating nucleic acid as a breast cancer biomarker. After a large amount of literature research, 96 miRNAs that are differentially expressed in breast cancer tissues and body fluids, or have functional verification at the level of breast cancer cell lines are selected as candidate genes for research; in particular, the present invention studies the relationship between circulating miRNAs and breast cancer Prognostic relationship of long-term follow-up population, 182 cases of patients based on binary regression analysis of machine training, revealed that 2‑10 miRNA combinations have a distinguishing effect on the prognosis of breast cancer patients, multivariate Cox regression analysis showed that 2‑7 miRNA combinations The combination is an independent risk factor for disease-free survival and distant disease-free survival in breast cancer patients, especially in patients with negative lymph nodes, the combination of 4-5 miRNAs has a better predictive effect on the survival of high- and low-risk breast cancer patients. This study opens up the possibility of using circulating miRNAs as tumor markers.

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 microRNAs (miRNAs) in breast cancer prognosis; more specifically, the present invention relates to the application of a combination of 2-10 different miRNAs in breast cancer prognosis, including based on the 2- A combination of 10 different miRNAs was used to prepare a kit for breast cancer prognosis, a microarray, and a method for predicting breast cancer prognosis using the kit and microarray. Background technique [0002] Breast cancer is the most common cancer in women worldwide, accounting for 23% of all cancers, and is also the leading cause of cancer death in women. According to the statistics of the World Health Organization in 2012 (GLOBOCAN 2012), compared with 2008, the global incidence of breast cancer has increased by 20% and the mortality rate has increased by...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/118C12Q2600/166C12Q2600/178
Inventor 不公告发明人
Owner KUNSHAN RNAI INST
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