Recombinant expression of hepatoma associated gene and its protein function.
10.3969/j.issn.1672-7347.2011.07.013
- Author:
Jie SONG
1
;
Yanhong LIU
;
Yalin LI
;
Guancheng LI
Author Information
1. Cancer Research Institute, Central South University, Changsha 410078, China.
- Publication Type:Journal Article
- MeSH:
Base Sequence;
Cell Proliferation;
drug effects;
Escherichia coli;
genetics;
metabolism;
Genes, Neoplasm;
physiology;
Genetic Vectors;
Hep G2 Cells;
Humans;
Maltose-Binding Proteins;
biosynthesis;
genetics;
pharmacology;
Molecular Sequence Data;
Neoplasm Proteins;
biosynthesis;
genetics;
pharmacology;
Open Reading Frames;
genetics;
Recombinant Fusion Proteins;
biosynthesis;
genetics;
pharmacology
- From:
Journal of Central South University(Medical Sciences)
2011;36(7):655-661
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To recombinant express hepatoma associated gene(HTA) and pre-test the function of HTA to determine the role of HTA in the development of liver cancer.
METHODS:HTA338-616 was amplified from HepG2 cells and cloned into the prokaryotic expression vector pET21a(+)-MBP. The proteins MBP and MBP-HTA were induced, purified by His-tag magnetic bead purification kit and identified by Western blot and ELISA. HepG2 cells were stimulated with MBP or MBP-HTA proteins. MTT assay and colony formation assay were employed to examine the proliferation of these cells and the changes of cell cycle distribution were determined by flow cytometry.
RESULTS:The prokaryotic expression plasmid pET21a(+)-MBP-HTA was successfully constructed. We got a 52 kD purified purpose protein.The proliferation of HepG2 cells stimulated with MBP-HTA was significantly higher than those stimulated with MBP and negative controls. HepG2 cells stimulated with MBP-HTA showed significant decrease fraction in G1 phase and increase fraction in S phase, and the cell proliferation was enhanced.
CONCLUSION:HTA protein can significantly promote the proliferation of HepG2 cells, which may be related to the promotion of G1 phase to S phase.
