A group of mutants suitable for classification and determination of patient prognosis in liver cancer histology

A technology for prognosis and mutation of liver cancer, applied in biochemical equipment and methods, analytical materials, scientific instruments, etc., can solve problems such as unclear effect

Active Publication Date: 2015-03-25
CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to effectively treat and prevent tumors (such as liver cancer), people have paid more and more attention to the early diagnosis and prognosis of tumors, but it is still unclear whether LLGL1 plays a role in the occurrence and development of liver cancer

Method used

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  • A group of mutants suitable for classification and determination of patient prognosis in liver cancer histology
  • A group of mutants suitable for classification and determination of patient prognosis in liver cancer histology
  • A group of mutants suitable for classification and determination of patient prognosis in liver cancer histology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0181] Example 1 Detection of LLGL1 mutation in liver cancer

[0182] Using the total RNA extraction reagent Trizol (Invitrogen Company) according to the operating instructions, extract 78 pairs of patients' liver cancer and paracancerous tissues and 5 strains of liver cancer cell lines (hepG2, huh7, SKhep1, SMMC-7721, BEL-7404, from the Chinese Academy of Sciences According to the total RNA, the total RNA was reacted with RT at 25°C for 5 minutes, 42°C for 60 minutes, and 70°C for 15 minutes. Four primers (P1: 5'-GGAATTCCGCAAGATGATGAAGTTTCG-3' (SEQ ID NO: 13) and P2: 5'-GACTGGCGCAGTGACTTCTTG-3' (SEQ ID NO: 14); P35'-ACTCTTCACCCCAATGACT-3' (SEQ ID NO: 15) and P45'-GCTCTAGAGATGCAACAGGCGTGGGCCT-3' (SEQ ID NO :16)) (eg figure 1 Shown in A) LLGL1 was amplified by two-stage PCR, and the cDNA of these samples was obtained from the reaction, the amplified PCR products were detected by 1% agarose gel, and the recovered PCR products were sequenced and analyzed (Boya Company). The in...

Embodiment 2

[0193] Example 2 Preparation of Three LLGL1 Mutants

[0194] Using primers 5'-CGGAATTCCAAGATGATGAAGTTTCGGTTC-3' (SEQ ID NO: 7) and 5'-GCTCTAGAT GATGGCCACGCTGACCGCC-3' (SEQ ID NO: 8), the cDNA of 7721 cells was amplified by PCR, and the amplified product was double-treated by EcorI enzyme and XbaI enzyme. After digestion, clone on the plasmid pEGFP-C1 (purchased from Invitrogene Company) to obtain the recombinant plasmid pEGFP-C1-7721 ( Figure 4 LLGL1-7721-EGFP in ). Two pairs of primers P1: 5'-GGAATTCCGCAA GATGATGAAGTTTCG-3, (SEQ ID NO: 9) and P2: 5'-GACTGGCGCAGTGACTTCTTG-3' (SEQ ID NO: 10); P3 5'-ACTCTTCACC CCAATGACT-3' (SEQ ID NO: 10) were used respectively. ID NO: 11) and P4 5'-GCTCTAGAGATGCAACAGGCG TGGGCCT-3' (SEQ ID NO: 12), PCR amplified 37k and 128k abnormal transcripts in two patient specimens respectively, and the amplified products were double-passed by EcorI enzyme and SmaI enzyme Digest the 5' (P1 / 2 amplification product) end and the 3' (P3 / P4 amplification prod...

Embodiment 3

[0196] Embodiment 3 detection kit

[0197] A liver cancer prognosis detection kit is prepared, wherein the following primer pair that can amplify the nucleotides from the 92nd to 1627th bases of SEQ ID NO: 1 is deleted: 5'-ACTCTTCACCCCAATGACT-3' (SEQ ID NO: 11); 5'-GCTCTAGAGATGCAACAGGCG TGGGCCT-3' (SEQ ID NO: 12).

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Abstract

The invention discloses a diagnosing method and detecting agent box of tumour susceptibility or prognosis, which comprises the following steps: detecting individual LLGL1 gene, transcription and or protein to compare normal LLGL1 gene, transcription and or protein; displaying difference higher than normal personal group and or tumour patient.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular, the invention relates to a group of LLGL1 variants, which are suitable for the histological grading of liver cancer and the judgment of patient susceptibility and / or prognosis. Background technique [0002] Hepatocellular carcinoma (hepatocellular carcinoma, HCC) is one of the common malignant tumors. The incidence of liver cancer ranks fifth among malignant tumors worldwide (>500,000 / year), and the mortality rate of liver cancer is high, ranking first in cancer deaths. Three major reasons (Block, T.M. et al., Molecular viral oncology of hepatocellular carcinoma. Oncogene 22(33):5093-107.2003). This has a lot to do with the pathogenesis of liver cancer. The onset of liver cancer is usually hidden, the early diagnosis rate is low, the clinical treatment effect is poor, and the five-year survival rate after diagnosis is less than 5%. The pathogenesis of liver cancer is currently ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68G01N33/574G01N33/68
Inventor 陈正军芦雪峰
Owner CENT FOR EXCELLENCE IN MOLECULAR CELL SCI CHINESE ACAD OF SCI
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