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目的探讨RNA干扰技术抑制蛋白激酶CK2α表达后,喉癌细胞Hep-2对顺铂敏感性的变化。方法构建蛋白激酶CK2α特异性siRNA表达质粒psiRNA-hH1neo-CK2α和非特异性表达质粒psiRNA-hH1neo-cont,脂质体转染法转染Hep-2细胞。采用逆转录聚合酶链反应(RT-PCR)、Western blot技术检测转染后Hep-2细胞蛋白激酶CK2α mRNA和蛋白表达。设立正常对照组、顺铂(5μg/ml)组、psiRNA-hH1neo-CK2浕组和psiRNA-hH1neo-CK2α+顺铂(5μg/ml)组,应用MTT法检测转染后Hep-2细胞对顺铂敏感性的变化。结果psiRNA-hH1neo-CK2α特异性地抑制了Hep-2细胞中蛋白激酶CK2α mRNA和蛋白的表达(P<0.05)。顺铂组(5μg/ml)及psiRNA-hH1neo-CK2浕组Hep-2细胞的增殖能力分别为正常对照组的(55.32±4.68)%和(49.68±5.33)%(P<0.05),但均高于二者联合组(26.12±5.51)%(P<0.05),转染psiRNA-hH1neo-CK2浕后,Hep-2细胞对顺铂敏感性增加了2.12倍。结论RNA干扰技术抑制蛋白激酶CK2浕表达能增加喉癌细胞Hep-2对顺铂的敏感性。
Objective To investigate the changes of cisplatin sensitivity of Hep-2 cells treated by RNA interference (RNAi) to inhibit protein kinase CK2α expression. Methods The psiRNA-hH1neo-CK2α specific protein kinase CK2α expression vector and the psiRNA-hH1neo-cont plasmid were constructed and transfected into Hep-2 cells by lipofectamine. Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect the protein and protein expression of CK2α in Hep-2 cells. The normal control group, cisplatin (5μg / ml) group, psiRNA-hH1neo-CK2 group and psiRNA-hH1neo-CK2α + cisplatin (5μg / ml) group were used to detect the transfection of Hep- Changes in platinum sensitivity. Results psiRNA-hH1neo-CK2α specifically inhibited protein kinase CK2α mRNA and protein expression in Hep-2 cells (P <0.05). The proliferation of Hep-2 cells in cisplatin group (5μg / ml) and psiRNA-hH1neo-CK2 group were (55.32 ± 4.68)% and (49.68 ± 5.33)% Compared with the combined control group (26.12 ± 5.51)% (P <0.05), the sensitivity of Hep-2 cells to cisplatin was increased by 2.12 times after transfected with psiRNA-hH1neo-CK2p. Conclusions RNA interference can inhibit the sensitivity of Hep-2 cells to cisplatin in inhibiting CK2 expression.