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Updated: May 29, 2026
Cell Type-specific Gene Expression Profiling in the Mouse Liver
Published on: September 17, 2019
[Recombinant expression of hepatoma associated gene and its protein function]
Jie Song1, Yanhong Liu, Yalin Li
1Cancer Research Institute, Central South University, Changsha 410078, China.
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.
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