Influence of folate on fragile histidine triad gene expression,cell proliferation and apoptosis in cervical cancer cell lines
10.3760/cma.j.issn.0254-6450.2014.05.022
- VernacularTitle:叶酸对脆性组氨酸三联体基因表达的影响及对宫颈癌细胞增殖凋亡的作用
- Author:
Chenfei GAO
1
;
Huijie KANG
;
Lixia BAI
;
Ling DING
;
Juan XU
;
Tingting WU
;
Lan BAI
;
Jintao WANG
Author Information
1. 山西医科大学公共卫生学院流行病学教研室
- Keywords:
Cervical cancer cells;
Folate;
Fragile histidine triad gene;
Expression
- From:
Chinese Journal of Epidemiology
2014;(5):569-572
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of folate on cell proliferation and apoptosis as well as on DNA methylation,expression of mRNA and protein of fragile histidine triad(FHIT)gene in cervical cancer cells. Methods Cervical cancer cell lines including CaSki(HPV16-positive)and C33A(HPV-negative)were cultured in vitro with different folate concentrations. Cell proliferation and apoptosis were determined by viable cell counting and flow cytometry while FHIT gene DNA methylation was used with methylation specific PCR(MSP). Both gene expression of FHIT protein and mRNA were detected by Western blot and Real-time PCR,respectively. Results Folate could inhibit the proliferation and promote the apoptosis in two kinds of cervical cancer cells. The number of viable cells decreased (C33A:r=0.98,P<0.001;CaSki:r=0.98,P<0.001) and the apoptosis rate increased(C33A:r=0.98,P<0.001;CaSki:r=0.99,P<0.001)along with the increase of folate concentration. FHIT gene DNA methylation showed all positive at the folate concentration levels of 1μg/ml and 10μg/ml,partially positive at 100μg/ml and 250μg/ml,but negative at 500μg/ml and 1 000μg/ml in both C33A and CaSki cells. Comparing with the control group,the mRNA or protein relative expression levels of FHIT gene in different folate concentrations were statistically significant in two kinds of cells,and showing that the FHIT gene mRNA expression(C33A:r=0.96,P<0.001;CaSki:r=0.94,P<0.001)and protein expression (C33A:r=0.96,P<0.001;CaSki:r=0.97,P<0.001) both increased along with the increase of folate concentration. Conclusion Our findings indicated that adequate folate seemed to be able to effectively inhibit the proliferation of cervical cancer cells and facilitate their apoptosis in vitro,so would reverse the aberration mRNA and protein expression of FHIT gene.