曲美替尼在卵巢癌中对PAX8的表达作用

Effects of Trametinib on PAX8 expression in ovarian cancer

目的探究MEK抑制剂—曲美替尼在卵巢癌中对PAX8表达的作用。方法应用基因富集分析(GSEA)检测卵巢癌细胞经曲美替尼处理后相关基因表达的差异,蛋白免疫印迹法、免疫荧光染色和实时荧光定量PCR检测曲美替尼对卵巢癌细胞PAX8蛋白表达和PAX8 mRNA的影响;结晶紫染色法检测曲美替尼对卵巢癌细胞增殖的影响;蛋白免疫印迹法、实时荧光定量PCR和CCK8实验检测干扰KRAS突变卵巢癌细胞ELK1基因后PAX8表达及细胞增殖的变化。结果GSEA结果显示,曲美替尼使卵巢癌细胞的KRAS基因和ELK1基因表达下调(P<0.01);实时荧光定量PCR结果显示,曲美替尼能抑制KRAS突变卵巢癌细胞中PAX8的转录(P<0.001);结晶紫染色法结果显示,曲美替尼处理后的KRAS突变卵巢癌细胞的增殖能力降低;蛋白免疫印迹法结果显示,曲美替尼能抑制KRAS突变卵巢癌细胞PAX8基因和ELK1基因表达(P<0.001),干扰KRAS突变卵巢癌细胞ELK1基因能抑制PAX8基因的表达(P<0.001);CCK8结果显示,干扰ELK1基因表达可抑制KRAS突变卵巢癌细胞增殖(P<0.05)。结论曲美替尼在KRAS突变的卵巢癌中通过ELK1抑制PAX8表达及细胞增殖。

Objective To investigate the effects of Trametinib,a MEK inhibitor,on PAX8 expression in ovarian cancer.Methods The expression changes of related genes in ovarian cancer cells treated with Trametinib were detected with gene set enrichment analysis(GSEA).The effects of Trametinib on the protein and mRNA expressions of PAX8 in ovarian cancer cells were detected with Western blotting,immunofluorescence staining and real-time quantitative PCR.The effects of Trametinib on the proliferation of ovarian cancer cells was detected with crystal violet staining.The change of PAX8 expression and cell proliferation in KRAS mutant ovarian cancer cells transfected with si-ELK1-RNA were detected with Western blotting,real-time quantitative PCR and CCK8 experiment.Results GSEA showed that Trametinib inhibited KRAS and ELK1 genes in ovarian cancer cells(P<0.01).Real-time quantitative PCR showed that Trametinib inhibited PAX8 transcription in KRAS mutant ovarian cancer cells(P<0.001).Crystal violet staining showed that Trametinib reduced the proliferation of KRAS mutated ovarian cancer cells.Western blotting showed that Trametinib inhibited the expression of PAX8 and ELK1 genes in KRAS mutant ovarian cancer cells(P<0.001),and interference with ELK1 gene inhibited the expression of PAX8 gene in KRAS mutant ovarian cancer cells(P<0.001);The results of CCK8 showed that interfering with ELK1 gene expression inhibited the proliferation of KRAS mutant ovarian cancer cells(P<0.05).Conclusion Trametinib inhibits PAX8 expression and cell proliferation through ELK1 gene in KRAS mutant ovarian cancer.