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Kit for carrying out early warning and screening on liver cancer

An early warning and kit technology, applied in the field of molecular biology, can solve the problems of unclear reasons for the down-regulation of ELF expression, and achieve the effects of saving DNA amount and time consumption, good application prospects, and high sensitivity

Active Publication Date: 2016-03-23
SICHUAN UNIV
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Problems solved by technology

It has been reported in the literature that the tumor suppressor gene ELF is expressed in normal liver tissue, while its expression is down-regulated in liver cancer stem cells and liver cancer tissues, and 40% of mice with knockout ELF gene spontaneously develop liver cancer, indicating that ELF plays an important role in the early stage of liver cancer. However, the reason for the down-regulation of ELF in liver cancer tissue has not been clarified

Method used

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  • Kit for carrying out early warning and screening on liver cancer
  • Kit for carrying out early warning and screening on liver cancer
  • Kit for carrying out early warning and screening on liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1 Composite amplification methylation-specific PCR capillary electrophoresis detection method test

[0059] 1. Specificity

[0060] K562 cells were used as LINE1 positive control, Hep3B cells were used as SOCS3 positive control, placental DNA modified by Sssl methyltransferase (purchased from NBI Company) was used as positive control for the methylation primers of RASSF1A, p16, SFRP1 and ELF four genes , while normal human liver tissue DNA was used as a positive control for unmethylated primers. The above six genes were all modified by bisulfite and subjected to multiple amplification methylation-specific PCR. see results figure 1 , 2 , 3, 4, 5, where figure 1 In order to use placental DNA modified by SssI methyltransferase and bisulfite as a template, 5-gene methylation primers were used for multiple amplification methylation-specific PCR, and capillary electrophoresis was used to detect bands. figure 2 In order to use normal human liver tissue DNA modifi...

Embodiment 2

[0065] Example 2 Multiple amplification of methylation-specific PCR and related detection of RASSF1A, p16, SFRP1, LINE1, ELF and SOCS3 gene promoter regions in human liver cancer, liver cirrhosis, benign liver lesions, and normal human plasma DNA

[0066] Tumor suppressor genes RASSF1A, p16, SFRP1, ELF and SOCS3 were expressed in normal human plasma, but were down-regulated in plasma and liver cancer tissues of liver cancer patients, while oncogene LINE1 was in contrast. From the perspective of DNA methylation, the present invention uses efficient and convenient multiplex amplification PCR technology and highly sensitive PAGE silver staining and capillary electrophoresis detection technology to explore the CpG islands in the promoter regions of RASSF1A, p16, SFRP1, LINE1, ELF and SOCS3 gene promoters The relationship between the methylation status of HCC and HCC lays an experimental foundation for further analysis of the methylation status of the gene RASSF1A, p16, SFRP1, LINE1...

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Abstract

The invention belongs to the field of molecular biology (tumor screening), particularly relates to a technology for carrying out early warning and screening on liver tumor, and aims to solve the technical problem to provide a new choice for early prediction of the liver tumor. The technical scheme of the invention is that a kit for carrying out early warning and screening on liver tumor comprises a primer which is used for amplifying the following genes of an RASSF1A (Ras-association Domain Family 1A) gene of which a promoter is in a CpG island methylation state, a p16 gene, ELF (Embryonic Liver Fodrin), an SOCS (Suppressors of Cytokine Signaling)-3 gene, an SFRP1 (Secreted Frizzled-Related Protein 1) gene and a genome repetitive sequence LINE1 (Long Interspersed Nuclear Elements 1). According to the kit disclosed by the invention, a new effective inspection method is provided for carrying out early warning and screening on early liver tumor and liver cirrhosis; the cost is low, the use is very convenient, the flexibility is high, the specificity is good, and the application prospect is very good.

Description

technical field [0001] The invention belongs to the field of molecular biology (tumor screening), and specifically relates to a technology that can be used for early warning and screening of liver cancer. Background technique [0002] Human primary hepatocellular carcinoma (hepatocellular carcinoma, HCC) refers to the cancer of liver cells or intrahepatic cholangiocytes. and esophageal cancer. There are about 120 million hepatitis B surface antigen carriers in my country, and about 3 million people may develop from acute hepatitis B to chronic hepatitis every year, and 300,000 people die from liver diseases, of which 120,000 people die from HCC. In my country, hepatitis B Hepatitis virus and hepatitis C virus infection are the most direct causes of the development of primary liver cancer (HCC). development process. Among the newly discovered liver cancer patients in the world every year, 42.5% of them occur in China, so China is a big country with liver cancer. Early surgi...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/154C12Q2600/158C12Q2600/16
Inventor 覃扬黄元魏玲孙爱民李波孙成均梁伟波刘秋英俞小琴何靖炀
Owner SICHUAN UNIV
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