摘要
目的通过抑制宫颈癌细胞株Siha中ΔNp63的表达,探讨ΔNp63对宫颈癌细胞增殖及凋亡的影响。方法将宫颈癌细胞株Siha分为实验组和对照组,实验组转染ΔNp63的特异性siRNA,对照组转染阴性对照siRNA。实时荧光定量PCR(QRT-PCR)及蛋白质印迹法检测Siha细胞转染前后ΔNp63基因在mRNA及蛋白水平的变化;CCK8法检测细胞增殖,流式细胞仪测定细胞凋亡率。结果实验组Siha细胞转染siRNA后,ΔNp63的mRNA表达为0.32±0.06,低于阴性对照组0.95±0.08(t=10.92,P<0.05)。实验组Siha细胞转染siRNA后,ΔNp63的蛋白表达为0.32±0.09,低于阴性对照组1.00±0.06(t=9.78,P<0.05)。实验组Siha细胞生长速度明显低于阴性对照组,实验组Siha细胞凋亡率为2.13±0.75,低于阴性对照组14.19±1.36(t=15.36,P<0.05)。结论人宫颈癌细胞株中存在ΔNp63的表达,特异性siRNA可下调宫颈癌细胞株Siha中ΔNp63基因的表达,对细胞的增殖具有负性调节作用,并能诱导细胞凋亡。
Objective To detect the effect of ΔNp63 on proliferation and apoptosis on human cervical cancer cells by suppressing the expression of ΔNp63 in cell line Siha. Methods Siha cell lines were divided into experimental group（ siRNA with transfected ΔNp63） and control group（ transfected negative controlled siRNA）. Real-time reverse transcription-polymerase chain reaction（ RT-PCR） and Western blot were used to detect the expressions of ΔNp63 in mRNA and protein before and after transfection. CCK8 was used to detect the proliferation of Siha cells and apoptosis was measured by flow cytometry. Results After Siha cells transfected siRNA in the experimental group,mRNA expression of ΔNp63 was 0. 32 ± 0. 06,which was lower than 0. 95 ± 0. 08 in the control group（ t = 10. 92,P 〈0. 05）,and the protein expression of ΔNp63 was 0. 32 ± 0. 09,which was lower than 1. 00 ± 0. 06 in the control group（ t = 9. 78,P 〈0. 05）. The growth speed of Siha cells was significantly lower in the experimental group than in the control group,and the apoptosis ratio of Siha cell in the experimental group（ 2. 13 ± 0. 75） was lower than that in the control group（ 14. 19 ± 1. 36）（ t = 15. 36,P〈 0. 05）. ConclusionΔNp63 exists in human cervical cancer cells. Specific siRNA can downregulate the expression of ΔNp63 in Siha,inhibit cell proliferation and induce apoptosis.
出处
《中国妇幼健康研究》
2016年第12期1469-1471,共3页
Chinese Journal of Woman and Child Health Research
