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HeLa Based Cell Free Expression Systems for Expression of Plasmodium Rhoptry Proteins
Published on: June 10, 2015
[Construction and package of the expression plasmid pAdEasy-1 system encoding the human papillomavirus 16 E7 gene and
Shu-zhen Dai1, Shou-fang Kong, Li-li Huang
1Department of Obstetrics and Gynecology, Affiliated Hospital of Medical College of Qingdao University, Qingdao 266003, China.
Objective:
To investigate the effect of human papillomavirus 16 E7 gene on cell cycle of cervical cancer HeLa cell, through construction and expression of human papillomavirus 16 E7 gene with adenovirus vector.
Methods:
Recombinant adenovirus which expressed E7 gene was constructed and packed. Flow cytometry (FCM) was used to detect the changes of cell cycle phase and cyclin D1 between cells infected and uninfected by recombinant adenovirus. The 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphennyltetrazolium bromide (MTT) colorimetric assay was used in the detection of the alteration of cell growth.
Results:
The recombinant adenovirus had stable efficiency of infection and E7 genes could be expressed stably. The result of MTT showed the multiplication of HeLa cells was accelerated from 0.27 +/- 0.03 before infection to 0.38 +/- 0.02 after infection (P < 0.01). FCM showed the number of cells in S phase increased from (26.0 +/- 0.4)% to (36.0 +/- 2.0)% at 12 hours and (49.9 +/- 4.2)% at 24 hours after infection (P < 0.05). Cyclin D1 expression was 22.4% before infection, and decreased to 55.2% after infection (P < 0.01).
Conclusions:
The recombinant adenovirus expressing E7 gene could infect target cells. E7 gene can influence cell-cycle of HeLa cells, which can be used to restrain cervical cancer.
