Deng Yuan-Run, Jiang Hui-Ping, Wu Lan-Fang, Chen Wei, Lin Dan, Guo Sui-Qun
Department of Obstetrics and Gynecology, Third Affiliated Hospital, Southern Medical University, Guangzhou 510630, China.E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2016 Aug 20;36(9):1226-1230.
To investigate the role of specificity protein 1 (Sp1) in regulating radiosensitivity of cervical cancer cell lines.
We analyzed Sp1 expression in 6 different cervical cancer cell lines (SiHa, HeLa, Caski, Me180, Ms751, and C33a) using Western blotting and real-time PCR. Clonogenic survival assay and curve fitting were used to assess the changes in radiosensitivity of Me180 cells transfected with lentivirus-mediated shRNA vector targeting sp1 and HeLa cells transfected with sp1 over-expression vector.
In the 6 cell lines tested, the cellular expression levels of Sp1 decreased gradually in the order of Me180, Caski, C33a, SiHa, Ms751, and HeLa. SP1 knockdown with lentivirus-mediated shRNA significantly lowered the survival rate of Me180 cells following radiation exposure (P<0.05), and obviously lowered the values of SF2, D0 and Dq but significantly increased α/β of the cells. Compared with the cells transfected with the mock vector, HeLa cells with sp1 over-expression showed a significantly increased survival following radiation exposure (P<0.05) with obviously increased values of SF2, D0 and Dq but significantly lowered α/β.
Silencing Sp1 can increase the radiosensitivity while Sp1 overexpression enhances the radioresistance of cervical cancer cell lines, suggesting an important role of Sp1 in radiotherapy for cervical cancer.
探讨特异性蛋白1(Sp1)在调节宫颈癌细胞系放射敏感性中的作用。
我们使用蛋白质免疫印迹法和实时荧光定量PCR分析了6种不同宫颈癌细胞系(SiHa、HeLa、Caski、Me180、Ms751和C33a)中Sp1的表达情况。采用克隆形成存活试验和曲线拟合来评估用慢病毒介导的靶向sp1的短发夹RNA载体转染的Me180细胞以及用sp1过表达载体转染的HeLa细胞放射敏感性的变化。
在所检测的6种细胞系中,Sp1的细胞表达水平按Me180、Caski、C33a、SiHa、Ms751和HeLa的顺序逐渐降低。用慢病毒介导的短发夹RNA敲低Sp1可显著降低Me180细胞受辐射后的存活率(P<0.05),并明显降低细胞的SF2、D0和Dq值,但显著增加α/β值。与转染空载体的细胞相比,过表达sp1的HeLa细胞受辐射后的存活率显著增加(P<0.05),SF2、D0和Dq值明显增加,但α/β显著降低。
沉默Sp1可增加放射敏感性,而Sp1过表达可增强宫颈癌细胞系的放射抗性,提示Sp1在宫颈癌放疗中起重要作用。