中华肿瘤杂志
中華腫瘤雜誌
중화종류잡지
CHINESE JOURNAL OF ONCOLOGY
2010年
5期
328-333
,共6页
周小羽%孙晋枫%何英华%张红宇%余坚%郭士成%蔡飏%胡夕春%朱景德
週小羽%孫晉楓%何英華%張紅宇%餘堅%郭士成%蔡飏%鬍夕春%硃景德
주소우%손진풍%하영화%장홍우%여견%곽사성%채양%호석춘%주경덕
乳腺肿瘤%DNA甲基化%CpG岛%氟尿嘧啶%化疗药物敏感性
乳腺腫瘤%DNA甲基化%CpG島%氟尿嘧啶%化療藥物敏感性
유선종류%DNA갑기화%CpG도%불뇨밀정%화료약물민감성
Breast neoplasms%DNA methylation%CpG islands%Fluorouraci%Chemosensitivity
目的 探讨10个基因的启动子CpG岛甲基化状态与乳腺癌细胞对5-氟尿嘧啶(5-Fu)敏感性的关系,确定与乳腺癌细胞株5-Fu敏感性相关的启动子CpG岛类基因.方法 采用四甲基偶氮唑蓝(MTT)法检测Bcap-37、T47D和ZR-75-30乳腺癌细胞株对5-Fu的敏感性.应用甲基化特异性PCR(MSP)方法,检测3株对5-Fu敏感性有差异的乳腺癌细胞株中BAG1、C110RF31、CBR1、CBR4、GJA1、FOXL2、IGFBP6、P4HA1、SRI和TYM基因的启动子CpG岛甲基化状态,并用实时定量PCR的方法检测PKSS21、LOX、IGFBP6、ABCC8和CHFR基因的稳定态mRNA表达的水平.结果 IGFBP6和FOXL2基因在对5-Fu敏感和耐药的乳腺癌细胞株中的甲基化状态有差异.除CHFR基因以外,PRSS21、LOX、IGFBP6和ABCC8基因的表达水平与启动子CpG岛甲基化状态有关,处于甲基化状态时表达水平低,而在处于去甲基化状态时表达水平高.结论 在敏感和耐药乳腺癌细胞株中存在甲基化状态差异的基因为IGFBP6和FOXL2基因,这为进一步开展DNA甲基化标志在乳腺癌化疗耐药中的应用研究提供了依据.
目的 探討10箇基因的啟動子CpG島甲基化狀態與乳腺癌細胞對5-氟尿嘧啶(5-Fu)敏感性的關繫,確定與乳腺癌細胞株5-Fu敏感性相關的啟動子CpG島類基因.方法 採用四甲基偶氮唑藍(MTT)法檢測Bcap-37、T47D和ZR-75-30乳腺癌細胞株對5-Fu的敏感性.應用甲基化特異性PCR(MSP)方法,檢測3株對5-Fu敏感性有差異的乳腺癌細胞株中BAG1、C110RF31、CBR1、CBR4、GJA1、FOXL2、IGFBP6、P4HA1、SRI和TYM基因的啟動子CpG島甲基化狀態,併用實時定量PCR的方法檢測PKSS21、LOX、IGFBP6、ABCC8和CHFR基因的穩定態mRNA錶達的水平.結果 IGFBP6和FOXL2基因在對5-Fu敏感和耐藥的乳腺癌細胞株中的甲基化狀態有差異.除CHFR基因以外,PRSS21、LOX、IGFBP6和ABCC8基因的錶達水平與啟動子CpG島甲基化狀態有關,處于甲基化狀態時錶達水平低,而在處于去甲基化狀態時錶達水平高.結論 在敏感和耐藥乳腺癌細胞株中存在甲基化狀態差異的基因為IGFBP6和FOXL2基因,這為進一步開展DNA甲基化標誌在乳腺癌化療耐藥中的應用研究提供瞭依據.
목적 탐토10개기인적계동자CpG도갑기화상태여유선암세포대5-불뇨밀정(5-Fu)민감성적관계,학정여유선암세포주5-Fu민감성상관적계동자CpG도류기인.방법 채용사갑기우담서람(MTT)법검측Bcap-37、T47D화ZR-75-30유선암세포주대5-Fu적민감성.응용갑기화특이성PCR(MSP)방법,검측3주대5-Fu민감성유차이적유선암세포주중BAG1、C110RF31、CBR1、CBR4、GJA1、FOXL2、IGFBP6、P4HA1、SRI화TYM기인적계동자CpG도갑기화상태,병용실시정량PCR적방법검측PKSS21、LOX、IGFBP6、ABCC8화CHFR기인적은정태mRNA표체적수평.결과 IGFBP6화FOXL2기인재대5-Fu민감화내약적유선암세포주중적갑기화상태유차이.제CHFR기인이외,PRSS21、LOX、IGFBP6화ABCC8기인적표체수평여계동자CpG도갑기화상태유관,처우갑기화상태시표체수평저,이재처우거갑기화상태시표체수평고.결론 재민감화내약유선암세포주중존재갑기화상태차이적기인위IGFBP6화FOXL2기인,저위진일보개전DNA갑기화표지재유선암화료내약중적응용연구제공료의거.
Objective To explore the relationship between the methylation status of CpG islands in the promoter region of 10 genes in breast cancer cells and their sensitivity to 5-fluouracil (5-Fu) , and to identify the genes responsible for the 5-Fu resistance in breast cancer. Methods Three cell lines (differently resistant to chemotherapy) were used in this study: Bcap-37 (IC50:289.77μg/ml), T47D (IC50:134.16μg/ml) and ZR-75-30 (IC50:4.20μg/ml). The methylation profile of 10 genes (BAG1,C11ORF31, CBR1, CBR4, GJA1, FOXL2, IGFBP6, P4HA1, SRI and TYMS) in the 3 breast cancer cell lines was determined by methylation specific PCR. The steady-state mRNAs of ABCC8, CHFR and IGFBP6 genes were quantified by real-time RT PCR analysis. Results Among the 10 genes, only genes IGFBP6 and FOXL2 displayed differential DNA methylation pattern between the 5-Fu-resistant and 5-Fu-sensitive cell lines. The mRNA expression level of genes PRSS21, LOX, IGFBP6, ABCC8 and CHFR was quantified by real-time RT-PCR analysis. Except for CHFR, the expression level of the other 4 genes was correlated with the methylation status of CpG islands, namely, a lower expression level with methylation status and a higher level with demethylation status. Conclusion The results of the present study have demonstrated that there are 8 genes with differential methylation status in chemosensitive and chemoresistant breast cancer cell lines,i. e. two genes more than the six genes we reported previously. Our findings provide both mechanistic insights for the drug resistance of breast cancer and the basis for further studies on potential application of the DNA methylation in this set of genes for prediction of chemosensitivity of breast cancer.
