Liver Cancer Cells ER Stress Model
When cells are disturbed by different environmental factors, the folding of peptide chains in endoplasmic reticulum will be blocked, and a large number of misfolded proteins will gather, so that cells are in a state of stress, which is called endoplasmic reticulum stress (ERS). The regulation mechanism of virus on endoplasmic reticulum stress and signal pathway is of great significance for the study of new antiviral drugs and new antiviral strategies. Hepatitis B virus (HBV) is a partially double stranded DNA virus with envelope. It is considered to cause ERS and is closely related to the occurrence and development of hepatocellular carcinoma. In order to further explore the relationship between hepatitis B virus and liver cancer, Creative Bioarray established a plasmid vector that can stably over express the HBx protein of hepatitis B virus.
Technology Principle
The expression of HBx can successfully induce ER stress in host cells.
HBV is a lysogenic virus of hepatotropic DNA virus family. Its genome has a circular structure. The product HBx (hepatitis B virus X protein) encoded by the X region of its open reading frame is a multifunctional regulatory protein. We cloned the coding region of HBx gene from hepatoma cell line by RT-PCR and inserted it into eukaryotic expression plasmid.
The vector can be transfected into human HepG2 cell line by liposome. After screening, the expression of HBx was verified by WB method, and the proliferation activity of cells was detected by MTT method.
RT-PCR was used to detect the expression of Er stress-related genes in cell lines. The results showed that the expression of BIP, PERK, ATF-6, XBP-1, and MANF were up-regulated.
Service Content
Construction, amplification and marketing of plasmids
Host cell culture and plasmid transfection services
Construction of human HepG2 cell line model overexpressing HBx protein
Endoplasmic reticulum stress model modeling effect and subsequent index detection service
Model Application
Different from the acute oxidative damage of cells caused by H2O2, the eukaryotic expression plasmid we provided can establish the endoplasmic reticulum stress model in hepatoma cell lines in a more mild and stable way
Through the analysis of ER stress related proteins BIP, PERK, ATF-6, XBP-1, and MANF, the model can be used to study the effect of hepatitis B virus on ER.
To further explore the relationship between hepatitis B virus and liver cancer.
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