摘要
目的探讨蛋白激酶CK2α特异性小干扰RNA(small interfering RNA,siRNA)对人喉癌细胞系Hep-2增殖和凋亡的影响。方法构建蛋白激酶CK2α特异性siRNA表达质粒psiRNA- hH1neo-CK2及非特异性表达质粒psiRNA-hH1neo-cont,分别用脂质体转染法转染Hep-2细胞。采用逆转录聚合酶链反应、Western Blot技术检测转染后蛋白激酶CK2αmRNA和蛋白表达,采用四甲基偶氮唑蓝法检测转染后Hep-2细胞的增殖情况,流式细胞术检测转染后对Hep-2细胞凋亡的影响。结果转染psiRNA—hH1neo—CK2载体后,Hep-2细胞蛋白激酶CK2αmRNA和蛋白表达均较非特异干扰组和空白组明显下降(P<0．05),Hep-2细胞生长缓慢(P<0．05),其细胞周期出现明显亚二倍体峰(P<0．05)。结论蛋白激酶CK2α特异性siRNA表达载体可以下调Hep-2细胞蛋白激酶CK2α的表达,抑制细胞增殖并诱导其凋亡。
Objective To assess the effect of small interfering RNA （siRNA） specific to protein kinase CK2α on proliferation and apoptosis of Hep-2 cell line. Methods siRNA expression plasmid psiRNA-hHlneo-CK2 specific to protein kinase CK2α and non-specific siRNA expression plasmid psiRNA- hHlneo-cont were constructed respectively, and then were transfected into Hep-2 cells by lipofectamine methods. Protein kinase CK2α mRNA and protein of the transfected cells were detected by reverse transcription polymerase chain reaction （RT-PCR） and Western Blot, respectively. Proliferation and apoptosis of the transfected cells were observed by methyl thiazolyl tetrazolium （MTT） method and flow cytometry （FCM）, respectively. Results Protein kinase CK20 mRNA and protein expressions were significantly decreased in the cells transfected with psiRNA-hHlneo-CK2（ P 〈 0. 05 ）. The Hep-2 cells grew slowly after transfected with psiRNA-hH1 neo-CK2（ P 〈 0. 05 ）. Obvious subdiploidy peaks were found in the cells transfected with psiRNA-hH1 neo-CK2 （P 〈 0.05 ）. Conclusions siRNA expression plasmid specific to protein kinase CK2α suppressed the protein kinase CK2α expression and the proliferation of Hep-2, and induced apoptosis of Hep-2 ceils.
出处
《中华耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2006年第8期617-621,共5页
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
基金
第四届教育部"高校青年教师奖"资助项目
关键词
喉肿瘤
癌
鳞状细胞
蛋白质丝氨酸苏氨酸激酶
逆转录聚合酶链反应
Laryngeal neoplasms
Carcinoma, squamous cell
Protein-serine-threornine kinases
Reverse transcriptase polymerase chain reaction
