Application of exosomes miRNA-485-3p and miRNA-885-5p as liver cancer diagnosis markers
A technology of diagnostic markers and exosomes, applied in the fields of biotechnology and medicine, can solve the problems of unsatisfactory early diagnosis indicators, and achieve the effects of economical and practical clinical development, accurate quantitative, and simple and fast methods.
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Embodiment 1
[0047] 1. Research objects: 21 patients with early-stage liver cancer from Beijing You'an Hospital affiliated to Capital Medical University from July 2018 to December 2020 were selected as the research group. There were 11 males and 10 females. The age ranged from 40 to 72 years old, with an average of (63.1±11.6) years old. The diagnostic criteria for early-stage liver cancer are based on the "Standards for the Diagnosis and Treatment of Primary Liver Cancer (2017 Edition)" issued by the National Health and Family Planning Commission. There were 28 patients in the chronic HBV infection group, including 15 males and 13 females, aged 42-69 years, with an average age of (54.2±8.7) years. The diagnostic criteria of chronic hepatitis B were based on the "Criteria for the diagnosis of chronic hepatitis B (2015 edition)" issued by the Chinese Society of Liver Diseases, Chinese Medical Association Infectious Diseases Branch. Thirty healthy volunteers diagnosed by the hospital's cli...
Embodiment 2
[0064] In the present invention, in order to study the plasma non-coding RNA molecular markers for the diagnosis of liver cancer, through literature research and the RT-PCR technology established in our laboratory, the plasma samples of 10 HBV carriers, liver cirrhosis patients, and liver cancer patients and 10 normal The control plasma exosome samples were detected, and the expression levels of 10 miRNAs were found to be different between the patient group and the normal group. After expanding the sample analysis of patients with early stage liver cancer, chronic HBV patients and healthy people, a candidate miRNA molecular marker group miRNA1323, miRNA-885-5p, miRNA-509-3p, miRNA-498, miRNA-485-3p, miRNA-516a-5p, miRNA-216B-5p, miRNA-194-3p, miRNA-455-3p, miRNA-498-3p. These 10 miRNA molecules were detected, and the detection results used bioinformatics analysis and medical statistical methods to find that the expression levels of miRNA molecules miR-485-3p and miR-885-5p bet...
Embodiment 3
[0066] The inspection methods used for the early screening of liver cancer are mainly various immunodetection technologies. Such technologies are mature and can perform accurate qualitative and quantitative detection of protein-based liver cancer markers. The expression profiles of miRNAs in different tumors are significantly different, so miRNA expression profiles can also be used as tumor diagnostic markers. In recent years, a lot of research has been done on the types of miRNAs and their expressions, aiming to explore the expression mechanism of miRNAs, in order to provide more effective markers for early clinical diagnosis of liver cancer.
[0067] The main steps of early diagnosis of liver cancer in the present invention are: plasma sample preparation; exosome extraction; exosome miRNA extraction; cDNA synthesis; cDNA product is subjected to fluorescence quantitative PCR reaction; 2-△△Ct analysis to assess the risk of liver cancer.
[0068] The beneficial effects of the ...
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