8-Br-cAMP联合槲皮素对人食管癌Eca-109细胞的逆转化作用

目的探讨8-Br-cAMP与槲皮素联合用药对人食管癌Eca-109细胞逆转化的作用。方法将培养的Eca-109细胞随机分为4组:①8-Br-cAMP组(Br):加终浓度为2×10-5mol/L8-Br-cAMP;②槲皮素组(Q):加槲皮素终浓度为43μmol/L;③8-Br-cAMP和槲皮素共同作用组(Br+Q):加终浓度为2×10-5mol/L8-Br-cAMP和终浓度为43μmol/L的槲皮素;④对照组(C):仅加DMEM培养液(含10%胎牛血清)。同时培养48h后分别对以上4组细胞制备细胞滴片及硝酸纤维素膜滴膜两种标本,进行Caspase-3和PCNA的免疫组化和免疫斑点印迹实验;应用完整细胞原位斑点印迹杂交技术检测c-myc、野生型p53(wtp53)、p16和EGFR的基因表达;并进行分化细胞/增殖细胞计数。结果8-Br-cAMP与槲皮素联合或单独应用均可上调Caspase-3免疫反应信号的表达,下调PCNA免疫反应信号的表达;同时上调wtp53和p16基因的表达,下调c-myc和EGFR基因的表达;且分化细胞的比率显著提高。结论8-Br-cAMP与槲皮素联合用药或单独用药均可通过调控人食管癌Eca-109细胞癌基因和抑癌基因的表达对Eca-109细胞起到生长抑制和促分化的作用。

Effects of 8-Br-cAMP combined with quercetin on reverse transformation of Eca-109 cells

Objective To study the effects of 8-Br-cAMP combined with guercetin on reverse transformation of Eca-109 cells. Methods The cultured Eca-109 cells were randomized into 4 groups: 8-Br-cAMP (Br) group, quercetin (Q) group, Br+Q group and control (C) group treated with on any drugs, each group cells were cultured for 48h. Two kinds of specimens including cell slide and NCM specimens were prepared and Caspase-3-IR and PCNA-IR were detected by immunohistochemistry and the immunodot blot technique. Besides, the gene expression of c-myc, wtp53, p16 and EGFR was detected by RNA dot blot. Results When compared with C group, Caspase-3-IR was increased and PCNA-IR was decreased; the gene expression of wtp53 and p16 was up-regulated, that of c-myc and EGFR was down-regulated in each drug-treated group. Conclusion The gene expression of oncogenes and anti-oncogenes may be involved in reverse transformation of Eca-109 cells induced by Br combined with Q or any drug alone. The effects of Br combined with Q are better than that of Br or Q alone in Eca-109 cell differentiation.