[Antisense targeting to human papillomavirus 18 E6/E7 affects the proliferation and apoptosis of human cervical carcinoma: an in vitro experiment with HeLa cells].
Zhonghua Yi Xue Za Zhi
; 87(23): 1618-21, 2007 Jun 19.
Article
en Zh
| MEDLINE
| ID: mdl-17803852
ABSTRACT
OBJECTIVE:
To investigate the effect of eukaryotic fluorescent expression vector carrying antisense human papillomavirus (HPV) 18 E6/E7 on the growth and proliferation of human cervical carcinoma.METHODS:
The HPV18 E6E7 with the length of 716 bp was amplified by PCR, the PCR product was inversely inserted into the eukaryotic fluorescent expression vector pEGFP-C1 so as to construct the recombinant eukaryotic expression plasmid pEGFP-HPV18E6E7as (EGFP-18AS). Human cervical carcinoma cells of the line HeLa were cultured and randomly divided into 3 groups Group A transfected with the recombinant plasmid EGFP-18AS, Group B transfected with the blank plasmid pEGFP-C1, and Group C without transfection used as control group. The mRNA expression of HPV 18 E6/E7 in the HeLa cells was detected by RT-PCR and protein expression of HPV18 E6/E7 HPV 18 E6/E7 in the HeLa cells was detected by. Western blotting MTT assay was performed to dynamically monitor the surviving cells and the cell apoptosis was observed by flow cytometry and fluorescence microscopy.RESULTS:
The protein and mRNA expression levels of HPV18 E6/E7 in the HeLa cells transfected with HeLa/18AS were both remarkably lower than those in the HeLa cells transfected with blank plasmid and those of the control group. The numbers of surviving HeLa cells of Group A was significantly lower than those of Groups B and C (both P < 0.05). The phenomenon of arrest of G(1) phase was remarkable in Group A. The apoptotic rate of the cells of Group A was 47.21%, significantly higher than those of Groups B and C (14.18% and 3.36% respectively, both P < 0.05). An increased number of cells with chromosome condensation and fragmentation was found in Group A as compared with Groups B and C.CONCLUSION:
The recombinant pEGFP-HPV18E6E7as can effectively inhibit the growth and proliferation of human cervical carcinoma HeLa cells, and further induce the cell apoptosis. The antisense RNA technology is available and may provide a new way to gene therapy of the cervical carcinoma.
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Colección:
01-internacional
Base de datos:
MEDLINE
Contexto en salud:
6_ODS3_enfermedades_notrasmisibles
Problema de salud:
6_cervical_cancer
Asunto principal:
Proteínas Oncogénicas Virales
/
Apoptosis
/
Proliferación Celular
/
Proteínas de Unión al ADN
Límite:
Female
/
Humans
Idioma:
Zh
Revista:
Zhonghua Yi Xue Za Zhi
Año:
2007
Tipo del documento:
Article
País de afiliación:
China