摘要
目的利用慢病毒(lentivirus)介导的短发卡RNA(short hairpin RNA,shRNA)建立人食管鳞状细胞癌(esophageal squamous-cell carcinoma,ESCC,以下简称食管鳞癌)裸鼠移植瘤模型,观察锌指蛋白8(plant homeodomain finger protein 8,PHF8)对移植瘤生长的影响。方法分别将未感染慢病毒(空白对照组)、感染Nonsilencing-shRNA慢病毒(阴性对照shRNA组)和PHF8shRNA慢病毒(PHF8 shRNA组)的人食管鳞癌细胞系TE-1接种于裸鼠背部皮下,建立食管鳞癌裸鼠移植瘤模型。定期测量肿瘤体积,绘制肿瘤生长曲线。4周后处死裸鼠,定量PCR及Western blotting法检测肿瘤组织中PHF8的表达。结果 PHF8shRNA组较阴性对照shRNA组、空白对照组的裸鼠致瘤能力明显减弱,PHF8 mRNA和蛋白的表达水平显著降低。结论慢病毒介导的shRNA干扰能有效减少食管鳞癌裸鼠移植瘤中PHF8的表达,而降低PHF8的表达能抑制食管鳞癌移植瘤的生长。
Objective To establish nude mice models with implanted tumor of esophageal squamous-cell carcinoma cells, and to observe the effects of PHF8 gene on the growth of implanted tumor. Methods Nude mice were injected subcutaneously into their right posterior flank with human TE-1 esophageal squamous-cell carcinoma cells （ control group）, lentivirus mediated Non-silencing shRNA TE- l cells （Non-silencing shRNA group）, or lentivirns mediated PHF8 shRNA TE-1 cells （PHF8 shRNA group）, respectively. The size of implanted tumor was measured at regular intervals and the growth curve of tumor was portrayed. Animals were sacrificed at 4 weeks, and the silencing of PHF8 in tumor tissues was confirmed by real-time PCR and Western blotting. Results The tumorigenic effect on nude mice was weaker and lower in PHF8 shRNA group, as compared to those in Non-silencing shRNA group and control group. The expression of PHF8 mRNA and protein in PHF8 shRNA group was significantly decreased. Conclusion Lentivirns mediated shRNA effectively inhibited the expression of PHF8 in transplanted tumor of nude mice with human esophageal squamous-eell carcinoma cell lines, and the tumorigenic ability was significantly reduced.
出处
《首都医科大学学报》
CAS
北大核心
2016年第1期12-16,共5页
Journal of Capital Medical University
基金
国家自然科学基金(81302160
81272447)
国家消化系统疾病临床医学研究中心基金(2015BAI13B09)
北京市医院管理局青苗人才计划(QML20150507)~~
关键词
PHD锌指蛋白8
慢病毒
食管鳞状细胞癌
裸鼠移植瘤
PHD finger protein 8
lentivirus
esophageal squamous cell carcinoma
transplanted tumor in nude mice
