摘要
目的:观察DNA甲基化转移酶(DNMT)抑制剂5-氮杂-2-脱氧胞苷(5-Aza-CdR)联合组蛋白去乙酰化酶(HDAC)抑制剂苯丁酸钠(SPB)对荷卵巢癌细胞系SKOV3裸鼠上皮钙黏素(E-cad)表达的影响。方法:在已建立的卵巢癌裸鼠腹腔移植瘤模型上,分别应用及两者联合应用5-Aza-CdR、SPB,免疫组化法检测腹腔移植瘤中DNMT1、HDAC1及E-cad的表达变化情况;检测各组移植瘤E-cad基因启动子区5CpG岛甲基化状况及移植瘤E-cadmRNA和蛋白的表达情况。结果:(1)在对照组中,DNMT1和HDAC1的阳性表达较多,5-Aza-CdR能抑制DNMT1的表达,SPB能抑制HDAC1的表达,差异有统计学意义(P<0.05)。(2)5-Aza-CdR能够诱导E-cad基因去甲基化,使E-cadmRNA及蛋白恢复表达或者表达增强,联合应用5-Aza-CdR和SPB可更大程度上诱导甲基化的E-cad基因去甲基化而恢复mRNA和E-cad蛋白的表达。结论:5-Aza-CdR能够降低E-cad基因启动子区的甲基化,可恢复其表达;联合应用5-Aza-CdR和SPB可产生协同效应,对卵巢癌的治疗有重要作用。′′
Objective： To evaluate the effect of the DNA methyltransferase （DNMT） inhibitor 5-Aza -2＇-deoxycytidine （5-Aza-CdR） combined with histone deaeetylase （HDAC） inhibitor sodium 4-pbenylbutyrate （SPB） on the expression of epi- thelial cadherin （E-cad） of nude mouse model with ovarian cancer cell line SKOV3. Methods： Forty nude mice were implant- ed ovarian cancer cell line SKOV3 into abdominal cavity as the animal model. The models were randomly divided into four groups, 5-Aza-CdR group, SPB group, 5-Aza-CdR＋SPB group and control group. The expressions of DNMT1, HDACI and E-cad in the ovarian cancer xenografts were detected by immunohistochemical staining. The CpG island methylation status of E-cadherin promoter region in xenograft tumors were analyzed by methylation specific PCR （MSP）. The mRNA and the pro- tein expressions of E-cad were compared between four groups. Results： The positive expressions of DNMT1 and HDAC1 were higher in control group. It was found that 5-Aza-CdR significantly inhibited the expression of DNMT1, and SPB signifi- cantly inhibited the expression of HDAC1 （P 〈 0.05）. It was also found that 5-Aza-CdR can induce the demethylation of E-cad gene, which enhanced expressions of E-cad mRNA and protein. Combined with 5-Aza-CdR and SPB induced demeth- ylation of E-cad gene and restored the E-cad mRNA and protein expressions. Conclusion： 5-Aza-CdR can reduce the E-cad gene promoter methylation and restore E-cad mRNA and protein expression. 5-Aza-CdR combined with SPB can ac- complish synergetic effect and enhance the expression of methylated E-cad gene significantly, which play an important role in the treatment of ovarian cancer.
出处
《天津医药》
CAS
北大核心
2012年第6期582-585,I0004,共5页
Tianjin Medical Journal
基金
天津市自然科学基金项目(项目编号:08JCYBJC08200)
