CAJ | 학술논문

岩藻糖基转移酶Ⅳ(Fucosyltransferase Ⅳ，FUT4)在正常细胞中表达量很低，但其低表达的调控机制以及是否受其启动子甲基化调控并不十分清楚。文章采用Western blot、免疫荧光和Real-time PCR的方法检测正常人永生化表皮细胞系HaCaT细胞FUT4的表达，观察DNA甲基转移酶抑制剂5-aza-dC处理对FUT4表达的影响。应用甲基化特异性PCR方法分析HaCaT细胞中FUT4启动子甲基化状态。结果表明，HaCaT细胞中FUT4的表达水平明显低于人表皮鳞癌细胞A431和SCC12。5μmol/L的5-aza-dC处理72 h的HaCaT细胞，其FUT4 mRNA水平明显升高，并且与未经5-aza-dC处理的对照组相比，U引物扩增检测到的产物量增加，M 引物扩增检测到的产物量明显减少。这些结果表明，HaCaT细胞中 FUT4的低表达可能与其启动子区 CpG岛甲基化有关。
암조당기전이매Ⅳ(Fucosyltransferase Ⅳ，FUT4)재정상세포중표체량흔저，단기저표체적조공궤제이급시부수기계동자갑기화조공병불십분청초。문장채용Western blot、면역형광화Real-time PCR적방법검측정상인영생화표피세포계HaCaT세포FUT4적표체，관찰DNA갑기전이매억제제5-aza-dC처리대FUT4표체적영향。응용갑기화특이성PCR방법분석HaCaT세포중FUT4계동자갑기화상태。결과표명，HaCaT세포중FUT4적표체수평명현저우인표피린암세포A431화SCC12。5μmol/L적5-aza-dC처리72 h적HaCaT세포，기FUT4 mRNA수평명현승고，병차여미경5-aza-dC처리적대조조상비，U인물확증검측도적산물량증가，M 인물확증검측도적산물량명현감소。저사결과표명，HaCaT세포중 FUT4적저표체가능여기계동자구 CpG도갑기화유관。
The expression level of fucosyltransferase Ⅳ (FUT4) is low in normal cells. The mechanism underlying regulation of FUT4 expression in normal cells remains elusive. In this study, Western blot, immunofluorescence and real-time PCR were used to analyze FUT4 expression in the immortalized human keratinocytes cells HaCaT. Methyl-ated-specific PCR was used to investigate methylation status of FUT4 promoter. The results showed that the FUT4 expression level was significantly lower in HaCaT cells than squamous carcinoma cells A431 and SCC12. FUT4 mRNA expression was increased in HaCaT cells treated by 5-aza-dC (5 μmol/L), an inhibitor of DNA methyltrans-ferase. Furthermore, using the primers to amplify the methylated fragment yielded PCR products and no products were yielded by the primers to amplify the unmethylated fragment in HaCaT cells. Unmethylated PCR products were obtained in HaCaT cells treated by 5-aza-dC, while methylated PCR products were not detected. These results suggest that the lower expression of FUT4 in HaCaT cells may be correlated with the methylation of CpG island in FUT4 promoter.