摘要
目的研究靶向全长配对盒基因-2(PAX2)siRNA在诱导子宫内膜癌细胞增殖、凋亡中的作用。方法慢病毒载体介导针对PAX2基因的siRNA转染HEC-1A细胞,Western blot、激光共聚焦显微镜、Annexin-V/PI双标法及四甲基偶氮唑蓝(MTT)分别检测PAX2蛋白表达、细胞凋亡形态学改变、凋亡率及细胞增殖变化。结果 PAX2-siRNA转染后,HEC-1A细胞的PAX2蛋白表达明显降低(P<0.05);细胞早期凋亡率显著增加(22.07±2.17)%,与空载体转染组(1.12±0.37)%及未转染组(0.89±0.52)%比较,差异均具有统计学意义(均P<0.01);细胞生长速率显著减慢,细胞经Hoechst 33258荧光染色,见转染后细胞的胞核呈浓集固缩改变,而空载体转染组和未转染组细胞的胞核无凋亡形态学改变。结论 PAX2特异性siRNA能明显抑制PAX2蛋白在子宫内膜癌HEC-1A细胞中的表达,抑制细胞增殖并诱导HEC-1A细胞凋亡。
Objective To investigate the effect of PAX2-siRNA on the proliferation and apoptosis of human endometrial carcinoma cell line.Methods siRNA lentivirus vector specifically targeting PAX2 gene was transfected into HEC-1A cells.The expression of protein of PAX2 was detected by using Western blot.Cell apoptosis morphology and cell apoptosis were determined by FITC-Annexin-V/PI double staining and Hoechst 33258 staining.Cell viability was detected by using MTT assay.Results The PAX2 protein expression in HEC-1A cells was decreased by PAX2 corresponding siRNA-encoding lentivirus vector transfection（P0.05）.The early period apoptosis rate of HEC-1A cells transfected with PAX2-siRNA was（22.07±2.17）%,which was significantly different from that in the empty vector group [（1.12±0.37）%] and negative control group [（0.89±0.52）%]（P0.01）.A marked morphological change of apoptosis was observed in PAX2-siRNA transfected HEC-1A cells.After transfection for 48 h,cells viability was significantly decreased in HEC-1A cells.Conclusion siRNA targeting PAX2 can suppress PAX2-protein expression markedly,inhibit the cell growth and enhance apoptosis of HEC-1A cells.
出处
《华中科技大学学报（医学版）》
CAS
CSCD
北大核心
2012年第1期84-87,共4页
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong
