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目的 :探讨PKC -α和PKA -Ⅰ反义核酸 (asODN)对鼻咽癌CNE - 2Z细胞生长增殖的影响。方法 :分别用脂质体 (lipofectin ,LP)介导asODN转染鼻咽癌CNE - 2Z细胞。实验组 :① 0 0 1- 1 0 0 μmol/LPKC -αasODN ;②0 0 1- 1 0 0 μmol/LPKA -ⅠasODN ;③ 0 5 0 μmol/LPKC -αasODN +0 5 0 μmol/LPKA -ⅠasODN。对照组 :相应浓度的随机序列 (rODN)。用免疫组化法检测CNE - 2Z细胞PKC -α和PKA -Ⅰ的表达 ,MTT法检测其生长指数 (growthin dex ,GI) ,软琼脂克隆形成率检测其体外增殖能力。结果 :PKC -αasODN或 /和PKA -ⅠasODN均可阻断其相应PKC-α或PKA -Ⅰ的表达 (P <0 0 5 )。PKC -αasODN或PKA -ⅠasODN均能显著降低CNE - 2Z细胞GI和软琼脂克隆形成率 (P <0 0 5 ) ,且具有量效依赖关系。PKC -αasODN +PKA -ⅠasODN共同作用可使其GI和软琼脂克隆形成率非常显著降低 (P <0 0 1) ,无明显量效依赖关系 ,两者对CNE - 2Z生长抑制作用强于PKC -αasODN或PKA -Ⅰa sODN(P <0 0 5 ) ,对其软琼脂克隆形成率的抑制作用与PKC -αasODN或PKA -ⅠasODN无显著差异 (P >0 0 5 )。结论 :PKC -αasODN和PKA -ⅠasODN均可抑制CNE - 2Z细胞体外生长和增殖 ,两者具有协同作用。
Objective: To investigate the effect of PKC-α and PKA-Ⅰ antisense oligonucleotide (asODN) on the growth and proliferation of nasopharyngeal carcinoma CNE - 2Z cells. Methods: Nasopharyngeal carcinoma CNE - 2Z cells were transfected with asODN by lipofectin (LP). The experimental group: ① 0 0 1 -1 0 0 μmol / LPKC -αasODN; ② 0 0 1 -1 0 0 μmol / LPKA-ⅠasODN; ③ 0 0 0 0 0 μ mol / LPKC-α asODN +0 0 0 0 mol / LPKA-ⅠasODN. Control group: the corresponding concentration of random sequence (rODN). The expressions of PKC-α and PKA-Ⅰ in CNE - 2Z cells were detected by immunohistochemistry. The growth index (gl) and the rate of soft agar colony formation were detected by MTT assay. Results: PKC-αasODN or / and PKA-IasODN could block the expression of PKC-α or PKA-Ⅰ (P <0.05). Both PKC-αasODN and PKA-ⅠasODN could significantly reduce the formation rate of GI and soft agar in CNE - 2Z cells (P <0 05) with a dose-dependent manner. The combined effect of PKC-αasODN + PKA-ⅠasODN could significantly reduce the formation rate of GI and soft agar (P <0.01), and there was no obvious dose-effect dependence. αasODN or PKA-Ⅰa sODN (P <0 05). The inhibitory effect on soft agar colony formation was not significantly different from PKC-αasODN or PKA-IasODN (P> 0.05). CONCLUSION: Both PKC-αasODN and PKA-ⅠasODN can inhibit the growth and proliferation of CNE - 2Z cells in vitro, both of which have synergistic effects.