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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 investigate the effects of small interfering RNA (siRNA) targeting protein kinase CK2α on the proliferation and apoptosis of human laryngeal carcinoma cell line Hep-2. Methods The psiRNA-hH1neo-CK2 specific protein kinase CK2α expression plasmid and the psiRNA-hH1neo-cont plasmid were constructed and transfected into Hep-2 cells by lipofectamine 2000 respectively. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of protein kinase CK2αmRNA and protein after transfection by Western Blot. The proliferation of Hep-2 cells was detected by MTT assay and the transfection by flow cytometry After Hep-2 cells apoptosis. Results The mRNA and protein expressions of protein kinase CK2α in Hep-2 cells were significantly decreased (P <0.05) and the growth of Hep-2 cells was slower than that in non-specific siRNA group .05), the cell cycle was significantly sub-diploid peak (P <0.05). Conclusion The protein kinase CK2α-specific siRNA expression vector can down-regulate the expression of protein kinase CK2α in Hep-2 cells, inhibit cell proliferation and induce apoptosis.