王金桃,吴婷婷,白兰,丁玲,郝敏,王云.叶酸对DNMTl和MeCP2在宫颈癌细胞中表达调节的实验研究[J].中华流行病学杂志,2013,34(2):173-177 |
叶酸对DNMTl和MeCP2在宫颈癌细胞中表达调节的实验研究 |
Effect 0f foIate in modulating the expression of DNA methyltransferase 1 and methyl.CpGbingding protein 2 in cervical cancer eelI Iines |
收稿日期:2012-09-22 出版日期:2014-08-07 |
DOI:10.3760/cma.j.issn.0254-6450.2013.02.016 |
中文关键词: 叶酸 宫颈癌细胞 DNA甲基转移酶1 甲基化CpG岛结合蛋白2 |
英文关键词: Folate Cervical cancer cell lines DNA methyltransferase 1(DNMTl) Methyl.CpG.Bingding protein 2(MeCP2) |
基金项目:国家自然科学基金(30872166,81273157);山西省自然科学基金(2008011075-1) |
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中文摘要: |
目的探讨宫颈癌细胞中叶酸对DNA甲基转移酶1(DNMTl)和甲基化CpG岛结合蛋白2(MeCP2)表达的调节作用。方法采用体外实验研究方法,对宫颈癌细胞casl(i(HPVl6阳性)和C33A(HPV阴性)进行不同浓度(10、50、100、500、750、1000 I.tg/m1)叶酸干预,分别采用Western blot和real-time PCR方法检测两种细胞中DNMTl和MeCP2蛋白的表达量和mRNA水平。结果随着叶酸水平的升高,两种细胞的生长抑制率(C33A:r=0.984,Pr=0.978,P=0.002)和凋亡率(C33A:r=0.989,P<0.001;Caski:r=0.994,Pr=0.914,Pr=0.859,P=0.003)及MeCP2蛋白表达(C33A:r=0.830,P=0.005;Caski:r=0.981,P<0.001)均呈逐渐降低趋势,而DNMTl和MeCP2 mRNA的Ct比值在两种细胞中的变化均无统计学意义(P>O.05)。相同叶酸浓度下,DNMTl蛋白和mRNA表达水平在Caski细胞均较C33A细胞为高,而MeCP2蛋白和mRNA表达水平则在C33A细胞较Caski细胞普遍为高。结论补充叶酸可有效抑制宫颈癌细胞的增殖,促进凋亡,逆转DNMTl和MeCP2蛋白的异常表达;HPVl6感染与DNMTl功能异常对宫颈癌的发生可能有协同效应。 |
英文摘要: |
objective To explore the eriects of folate on the expression of DNA memyltransferase 1(DNMTl)and methyl.CpG.bingding protein 2(MeCP2)in cervical cancer eell lines.Methods Experimental study was carried out in vitro.Human cervical cancer cell lines,
including C33A eell with HPV negative and Caski eell with HPV 1 6 positive,were treated witll different concentration of folate.The expression of DNMTl and MeCP2 protein(bv Westem blot)and mRNA(by real.time PCR)were then detected in the two eell lines.Results It was found that supplement offolate was able to reduce the eell proliferation in C33A eell(r=0.984,P<0.001)and Caski eell(r=0.978,P=0.002),as well as induced the eell apoptosis(C33A:r=0.989,P<0.001;Caski:r=0.994.P<0.001).Results showed that the expression levels of DNMTl protein(C33A:r=-0.914,P<0.001;Caski:r=0.859,P=0.003)andMeCP2 protein(C33A:r=0.830,P=0.005;Caski:r=0.98l,P<0.001)decreased gradually with the increase of folate concentrations,but the expression of DNMTl and MeCP2 mRNA was not observed in Caski or C33A cell.When at the same levels of folate.the expression of DNMTl protein or mRNA was higher in Caski eell than in C33A cell.However.the expression of MeCP2 protein or mRNA was higher in C33A eell than in Caski cell.Conclusion Our finding indicated that adequate foleta could effectively inhibit the proliferation of
cervicaI cancer cells and facilitate their apoptosis in vitro.thus would reverse the aberration protein expression ofDNMTl and MeCP2.That there might be a synergistic action between HPVl6 iIlfection and parafunction ofDNMTl in cervical cancer.being noticed. |
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