Proliferation Of Human Cervical Cells Research Articles

Impacts of isoliquiritigenin on proliferation and apoptosis of human cervical cancer cells

Impacts of isoliquiritigenin on proliferation and apoptosis of human cervical cancer cells

Antisense targeting to human papillomavirus 18 E6/E7 affects the proliferation and apoptosis of human cervical carcinoma: an in vitro experiment with HeLa cells

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. 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. 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. 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.

Effect of Nicotine on Proliferation of Normal, Malignant, and Human Papillomavirus-Transformed Human Cervical Cells

Nicotine is concentrated in the cervical mucus of smokers relative to serum levels. In this experiment, the effect of nicotine on cellular proliferation of human ectocervical, endocervical, malignant, and human papillomavirus (HPV) 16 DNA-transformed cervical cell lines was studied. Ectocervical and endocervical cell lines were derived from benign hysterectomy specimens and cultured in keratinocyte growth medium. HPV 16 DNA-transformed cell lines were derived through transfection of ectocervical cells with cloned HPV 16 DNA. HPV-transformed lines and malignant cell lines established from three women with newly diagnosed cervical cancer were maintained in E-media with 5% fetal calf serum. Cells (2500-5000) were cultured in 96-well tissue culture plates with varying concentrations of nicotine (100 to 10 mg/ml) and proliferation was assessed 72 hr later with a semiautomated colorimetric assay. Experiments were performed three times and proliferation of nicotine-exposed cells was compared to unexposed cells with one-way analysis of variance. Nicotine, at 100 ug/ml to 10 μg/ml, significantly stimulated epithelial cell growth in two ectocervical and three HPV DNA-transformed cell lines (proliferation 118-180% of control, P < .05). Nicotine at 100 ng/ml to 10 μg/ml did not significantly alter proliferation of four endocervical, three malignant, and two other ectocervical cell lines. Toxic effects of nicotine (>50% inhibition of cellular proliferation) were noted between 100 μg/ml and 10 mg/ml and exceed the concentrations of nicotine reported in smoker's cervical mucus. These findings demonstrate that nicotine, in physiologically attainable concentrations, does not impair and occasionally enhances the proliferation of human cervical cells in vitro. The selective mitogenic effect noted among normal ectocervical and HPV-transformed ectocervical cells may relate to epidemiologic studies showing, among smokers, an increased risk of squamous cell carcinoma, and not adenocarcinoma, of the cervix.