| 高晨菲,康慧杰,白丽霞,丁玲,许娟,吴婷婷,白兰,王金桃.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[J].Chinese journal of Epidemiology,2014,35(5):569-572 |
| 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 |
| Influence of folate on fragile histidine triad gene expression，cell proliferation and apoptosis in cervical cancer cell lines |
| Received:November 05, 2013 |
| DOI：10.3760/cma.j.issn.0254-6450.2014.05.022 |
| KeyWord: 宫颈癌细胞 叶酸 脆性组氨酸三联体基因 表达 |
| English Key Word: Cervical cancer cells Folate Fragile histidine triad gene Expression |
| FundProject:国家自然科学基金（30872166，81273157）；山西省自然科学基金（2008011075-1） |
| Author Name | Affiliation | E-mail | | Gao Chenfei | Department of Epidemiology, School of Public Health，Shanxi Medical University, Taiyuan 030001, China | | | Kang Huijie | Department of Epidemiology, School of Public Health，Shanxi Medical University, Taiyuan 030001, China | | | Bai Lixia | Department of Epidemiology, School of Public Health，Shanxi Medical University, Taiyuan 030001, China | | | Ding Ling | Department of Epidemiology, School of Public Health，Shanxi Medical University, Taiyuan 030001, China | | | Xu Juan | Department of Epidemiology, School of Public Health，Shanxi Medical University, Taiyuan 030001, China | | | Wu Tingting | Department of Epidemiology, School of Public Health，Shanxi Medical University, Taiyuan 030001, China | | | Bai Lan | Department of Epidemiology, School of Public Health，Shanxi Medical University, Taiyuan 030001, China | | | Wang Jintao | Department of Epidemiology, School of Public Health，Shanxi Medical University, Taiyuan 030001, China | wangjt59@163.com |
|
| Hits: 3736 |
| Download times: 1231 |
| Abstract: |
| 目的 探讨叶酸对脆性组氨酸三联体（FHIT）基因DNA甲基化、mRNA与蛋白表达以及宫颈癌细胞增殖、凋亡的影响。方法 采取体外细胞实验方法，对两种宫颈癌细胞C33A（HPV阴性）与CaSk（iHPV16阳性）进行不同浓度叶酸干预，用活细胞计数和流式细胞术分别测定宫颈癌细胞的增殖和凋亡情况，采用甲基化特异性PCR（MSP）法测定FHIT基因启动子区CpG 甲基化状态，利用 Western blot 和 Real-time PCR 方法分别检测 FHIT 基因蛋白和 mRNA 的表达水平。结果 不同浓度叶酸对两种宫颈癌细胞具有抑制增殖和促进凋亡的作用。随着叶酸浓度增加，抑制增殖率逐渐上升（C33A：r＝0.98，P<0.001；CaSki：r＝0.98，P<0.001），细胞凋亡率升高（C33A：r＝0.98，P<0.001；CaSki：r＝0.99，P<0.001）。C33A与CaSki两种细胞在叶酸浓度为1 μg/ml和10 μg/ml时均呈现FHIT基因DNA甲基化阳性，100 μg/ml和250 μg/ml显示为部分阳性，500 μg/ml和1 000 μg/ml时均呈甲基化阴性。不同叶酸浓度组与对照组相比，两种细胞FHIT 基因的 mRNA 和蛋白相对表达量的差异均有统计学意义，随叶酸浓度的升高，FHIT mRNA（C33A：r＝0.96，P<0.001；CaSki：r＝0.94，P<0.001）及蛋白（C33A：r＝0.96，P<0.001；CaSki：r＝0.97，P<0.001）的表达量均呈上升趋势。结论 较高浓度叶酸可降低 FHIT 抑癌基因的 DNA 甲基化水平，逆转其在转录和功能水平的异常表达，对抑制宫颈癌细胞增殖、促进凋亡具有不可忽视的作用。 |
| English 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 1 μg/ml and 10 μg/ml，partially positive at 100 μg/ml and 250 μg/ml，but negative at 500 μg/ml and 1 000 μg/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 proteinexpression of FHIT gene. |
|
View Fulltext
Html FullText
View/Add Comment Download reader |
| Close |
|
|
|