Kits for early warning and screening of 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
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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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