摘要
本研究探讨叉状头/翅膀状螺旋转录因子(forkhead/winged helix transcription factor,Foxp3)过表达对人乳头状瘤病毒(human papilloma virus,HPV)感染阳性宫颈癌细胞增殖、凋亡及细胞免疫的影响,为阐明宫颈癌肿瘤免疫逃逸机制、肿瘤治疗及药物开发提供一定理论基础。利用脂质体介导转染方法,研究者建立Foxp3过表达HPV感染阳性SiHa细胞株,作为Foxp3过表达阳性细胞,同时利用pcDNA3.1空载体转染SiHa细胞株,作为本研究阴性对照,同时设置空白对照组,采用qRT-PCR方法及Western blotting法检测3组Foxp3mRNA及蛋白表达水平。采用ELISA法检测SiHa细胞中IL-2、IL-10水平,采用MTT实验检测SiHa细胞增殖能力,流式细胞术检测SiHa细胞周期及凋亡情况。结果显示过表达组Foxp3mRNA及蛋白表达水平明显高于阴性对照组及空白对照组,差异有统计学意义(P<0.05),阴性对照组及空白对照组Foxp3mRNA及蛋白表达水平相比较,差异无统计学意义(P>0.05);与阴性对照组及空白对照组比较,过表达组IL-2水平明显降低,IL-10水平明显升高;MTT结果显示:Foxp3过表达可促进SiHa细胞增殖;FCM检测显示,Foxp3过表达可明显抑制SiHa细胞凋亡,抑制G0/G1期阻滞。由此,Foxp3过表达可促进HPV感染阳性宫颈癌细胞增殖,抑制细胞凋亡。
This study aimed to investigate the effects of overexpression of forkhead/winged helix transcription factor（Foxp3）on proliferation,apoptosis and cell immunity of human cervical cancer cell line infected with human papilloma virus（HPV）to clarify the immune escape mechanism of cervical cancer,and to provide a theoretical basis for tumor treatment and drug development.Using liposome-mediated transfection,we established Foxp3 overexpressed HPV-positive SiHa cell line and using pcDNA3.1 empty vector transfected SiHa cell lines as negative control,and the blank control group was also set up.The mRNA and protein expression levels of Foxp3 in the three groups were detected by qRT-PCR and Western blotting.The levels of IL-2 and IL-10 in SiHa cells were detected by ELISA.The proliferation of SiHa cells was detected by MTT assay.The cell cycle and apoptosis of SiHa cells were detected by flow cytometry.The results showed that Foxp3 mRNA and protein expression levels were significantly higher in the overexpression group than in the negative control group and the blank control group,the pairwise differences were statistically significant（P〈0.05）,while the difference between the negative control group and the blank control group was non significant（P〉0.05）.Compared with the negative control group and the blank control group,the level of IL-2 in overexpression group was significantly decreased and the level of IL-10 was significantly increased.MTT results showed that overexpression of Foxp3 could promote SiHa cells proliferation.FCM showed that Foxp3 overexpression could significantly inhibit SiHa cell apoptosis and inhibit G0/G1 phase arrest.Thus,Foxp3 overexpression can promote proliferation,and inhibit apoptosis,of HPV-positive cervical cancer cells.
作者
马薇
张书勤
MA Wei;ZHANG Shu-qin(Huazhong University of Science and Technology Tongji Hospital Affiliated Pear Garden Hospital,Wuhan 430077,Chin)
出处
《现代免疫学》
CAS
CSCD
北大核心
2018年第4期276-280,共5页
Current Immunology
关键词
人乳头状瘤病毒
宫颈癌
叉状头/翅膀状螺旋转录因子
增殖
凋亡
human papilloma virus （HPV）
cervical carcinoma
forkhead/winged helix transcription factor
proliferation
apoptosis
