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Liver cancer circulating tumor cell marker and application thereof

A tumor cell and marker technology, applied in the field of biological detection, can solve the problems of missed detection of CTCs, and achieve the effects of high sensitivity, simple design, high specificity and sensitivity

Pending Publication Date: 2021-04-16
NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have shown that CTCs are one of the important driving factors of tumor recurrence and metastasis. Obviously, the dynamic biological process of EMT makes the markers of CTCs uncertain, and relying on epithelial markers alone to identify CTCs may lead to most Mesenchymal CTCs are missed, and these mesenchymal markers may have important clinical significance

Method used

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  • Liver cancer circulating tumor cell marker and application thereof
  • Liver cancer circulating tumor cell marker and application thereof
  • Liver cancer circulating tumor cell marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048]Example 1 Screening and verification of tumor cells and blood cells expressing lipid metabolic markers

[0049]Material and method

[0050](1) Cell strain

[0051]7 Liver Cell Cellular: MHCC97-L, MHCC97-H, Hep3B2.1-7 and MHCC97-LM3 Purchad in the Liver Cancer Research Institute, Zhongshan Hospital, Fudan University, SNU-182 is purchased in the Chinese Academy of Sciences, Hepg2, SK-Hep1 Divided by professor of Fuwei, Huang Ning graduate students (Southern Medical University);

[0052](2) Medium and fetal bovine serum

[0053]The fetal bovine serum, DMEM, RPMI-1640 is purchased in Guangzhou Jets Technology Co., Ltd. (GIBCO production), EMEM (containing NEAA), from Guangzhou Hao Road Biotechnology Co., Ltd. (ATCC production). DMEM was used to cultivate MHCC97-L, MHCC97-H, MHCC97-LM3 and SNU-182 cells, and 10% fetal bovine serum was added during culture; RPMI-1640 was used to cultivate HepG2 and SK-HEP1 cells; EMEM was used to cultivate Hep3B2.1 -7, add 15% fetal bovine serum during culture.

[00...

Embodiment 3

[0096]Example 3 Lipin metabolic markers for CTCS identification and layout in peripheral blood samples in tumor patients

[0097]1, materials and methods

[0098](1) Recruitment of patients with hepatocyte cancer and accelerated patients with hepatocarcinoma patients, while recruiting 5 patients with liver benign diseases (including chronic hepatitis, fatty liver, cirrhosis, etc.), and all patients Agree, 10 ml of peripheral blood was collected, and the expression of the number of CTCs and its lipid metabolism such as the above method (Example 2) was collected.

[0099](2) After removing plasma, use RosettesepTM The kit is negatively enriched CTCs. MRNA in situ lock hybridization and fluorescence label were performed in a CTCS fixed to the slide using Hoechst33342 cell nuclear dye and AOPB, Alb, and CD45, CTCs fixed to the slide, and the peripheral blood CTCs were detected according to the foregoing method (Method of Example 2). APOB and ALB marker signals to identify CTCs and are typed.

[010...

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Abstract

The invention belongs to the technical field of biological detection, and particularly relates to a liver cancer circulating tumor cell marker and application thereof. Circulating tumor cells (CTCs) are subjected to functional identification from the perspective of abnormal tumor lipid metabolism activity, and due to the fact that lipid metabolism related signal pathway molecules are relatively stable in expression in cancer progression, the detection omission of liver cancer circulating tumor cells can be avoided, so that the monitoring accuracy is improved, and the biological behaviors of CTCs can be better reflected. Meanwhile, the primer and the probe (reverse transcription primer and lock type probe) can be used for mRNA detection of liver cancer cells APOB and / or ALB, and have the characteristics of simple design, strong detection signal in in-situ hybridization, no interference and the like, and in-situ lock type probe hybridization experiments show that the primer and the probe can be used for CTCs identification of clinical liver cancer patient blood samples, and have the characteristics of high specificity and high sensitivity, and are not limited to the use of blood samples.

Description

Technical field[0001]The present invention belongs to the technical field of biological detection, and is specifically involved in a liver cancer circulating tumor cell marker and its application thereof.Background technique[0002]Circulatingtumor Cells (CTCs) is a rare cell from the in-situ solid tumor or the transfer tumor portion or from clinical diagnosis and treatment, and released to a rare cells of the blood circulation. In recent years, as one of the markers of "liquid biopsy" techniques expected to replace traditional tissue biopsy, CTCs have been reported and metastatic breast cancer, colorectal cancer, prostate cancer, small cell lung cancer, non-small cell lung cancer Transfer, recurrence, no progress, and total survival period, etc. The greatest advantage of "liquid biopsy" is to continuously sampling, minimally invasive, convenient, convenient, and can overcome traditional organizational biopsy difficult to sample, transfer stoves, and bleeding risk, etc., there is a br...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6841C12Q1/682
Inventor 陈书坤郑磊孙春霞何叶
Owner NANFANG HOSPITAL OF SOUTHERN MEDICAL UNIV