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Luc-GFP marked high-transitivity human hepatoma cell line and application thereof in in situ hepatoma model

A technology of human liver cancer cell lines and liver cancer cells, which is applied in the field of biomedicine, can solve problems such as the absence of human liver cancer cell lines, and achieve the effect of great application prospects

Active Publication Date: 2018-07-17
GUANGDONG PHARMA UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

[0005] There is no report on the successful construction of a human liver cancer cell line stably expressing luciferase (Luc) and green fluorescent protein (GFP).

Method used

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  • Luc-GFP marked high-transitivity human hepatoma cell line and application thereof in in situ hepatoma model
  • Luc-GFP marked high-transitivity human hepatoma cell line and application thereof in in situ hepatoma model
  • Luc-GFP marked high-transitivity human hepatoma cell line and application thereof in in situ hepatoma model

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Experimental program
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Embodiment 1

[0037] Example 1 Construction of human liver cancer cell line HCCLM3-Luc-GFP

[0038] 1. PCR amplification of luciferase gene

[0039] (1) According to the luciferase (Luc) gene sequence (SEQ ID NO: 3) on the psiCHECK-2 vector and the multi-cloning site on the lentiviral expression vector pCDH-CMV-MCS-EF1-GFP-T2A-Puro, design two specificity Primers:

[0040] P1: 5'-TGC TCTAGA ATGGAAGACGCCAAAAACATAAAGA-3' (SEQ ID NO: 1)

[0041] P2: 5'CGC GGATCC TTACACGGCGATCTTTTCCGCCCTTC-3' (SEQ ID NO: 2)

[0042] (Note: " in the P1 primer TCTAGA "is the XbaI restriction site,;" in the primer P2 GGATCC ” is the BamHI restriction site).

[0043] (2) Using the psiCHECK-2 vector as a template, primers P1 and P2 amplify the 1653bp luciferase gene sequence. Restriction endonuclease XbaI and BamHI sites were respectively introduced at both ends of the luciferase sequence. Primers were synthesized by Shanghai Yingwei Jieji Company.

[0044] The results showed that the psiCHECK-2 vector...

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Abstract

The invention discloses an Luc-GFP marked high-transitivity human hepatoma cell line. Hepatoma cells are taken as mother cells, a virus solution packaged with pCDH-CMV-Luc-EF1-GFP-Puro is transfectedthrough slow viruses, and then through puromycin screening, the high-transitivity human hepatoma cell line with stable GFP expression and luciferase activity is obtained. By using the Luc-GFP marked high-transitivity human hepatoma cell line, an in situ hepatoma model for a naked mouse is established through a cell suspension method and a tumor tissue block conversion method respectively, the tumor formation rate of the hepatoma model is 100% respectively, the rates of lung metastasis are 100% and 80% respectively, a foundation is laid for monitoring the growth of tumor in the level of livinganimals and evaluating the curative effect of hepatocellular carcinoma metastasis resistant medicine, and the cell line has greater application prospect.

Description

technical field [0001] The invention belongs to the field of biomedicine, and more specifically relates to a Luc-GFP-labeled highly metastatic human liver cancer cell line and its application in an orthotopic liver cancer model. Background technique [0002] Primary hepatocellular carcinoma (hepatocellular carcinoma, HCC, referred to as liver cancer) is a malignant tumor with the sixth highest incidence rate and the third highest mortality rate in the world. my country is a country with a high incidence of liver cancer. There are about 350,000 new cases of liver cancer every year, and about 320,000 people die from liver cancer every year. Invasion, metastasis and recurrence of HCC are one of the main causes of death in patients. Therefore, it is of great significance to explore the mechanism of liver cancer invasion, metastasis and recurrence, and to find drugs that can effectively inhibit the metastasis and recurrence of liver cancer. The establishment of a nude mouse mode...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12Q1/02A01K67/027
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/03C12N5/0693C12N2503/02C12N2510/00G01N33/5011G01N2500/10
Inventor 马艳
Owner GUANGDONG PHARMA UNIV
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