蛋白激酶C抑制剂诱导CNE-2Z细胞凋亡的研究

Study on the apoptosis in CNE-2Z cells induced by inhibitors of protein kinase C

目的和方法：采用蛋白激酶Ｃ（ＰＫＣ）催化区抑制剂ｓｔａｕｒｏｓｐｏｒｉｎｅ（ＳＴ）、调节区抑制剂ｓｐｈｉｎｇｏｓｉｎｅ（ＳＳ），诱导ＣＮＥ—２Ｚ细胞２４ｈ。结果：（１）ＳＴ、ＳＳ对细胞ＤＮＡ裂解的影响均随浓度增高而逐步增强，作用明显的终浓度分别为１×１０６ｍｏｌ／Ｌ和４×１０５；（２）核形态观察：诱导后部分细胞核变小，核浓缩及核碎裂；（３）ＤＮＡ琼脂糖电泳：诱导后的细胞有典型梯状ＤＮＡ条带；（４）流式细胞仪分析：诱导组Ｇ１期前均有ＤＮＡ亚二倍体峰，即凋亡峰。细胞周期改变：ＳＴ使Ｇ２期增加、Ｇ１、Ｓ期减少；ＳＳ使Ｓ期增加和Ｇ１期减少。结论：ＰＫＣ抑制剂可有效地促进ＣＮＥ－２Ｚ细胞凋亡，但与抑制剂剂量有关，细胞周期的改变可能在ＰＫＣ抑制剂诱导的细胞凋亡中起重要作用。

AIM and METHOD: CNE-ZZ cells werc induced for 24 hours by staurosporine (ST) acted on the catalytic domain of protein kinase C(PKC) and sphingosine (SS) inhibited on modulating domain of PKC respectively. RESULTS: (l) With improved concentration of SS and ST, the DNA fragment rates had increased. The marked effects of SS and ST to DNA fragment rates were at the concentration of 1 × 10 6 mol/L and 4× 10 5 mol/L and cultured for 24hrs,respectively. (2)Cell morphology: cells shrinkage, chromation condensation and nuclear fragmentation wcrc observed in treated groups. (3) Gelelectrophoresis ofDNA extracted from the trealed cells showed oligonucleosomal ladders.(4) Flow cytometery(FCM) analysis: hypodiploid DNA peaks(apoptotic peak) were seen in treated groups. Cell cycleanalysis showed that G 2 phrase increased, but both G 1 and S phrase decreased in ST treated groups, and S phrase increased, but G 1 phrase decreased in SS treated groups. CONCLUSION:PKC inhibitors effectively induced apoptosis, but DNA fragment rate was related to the does of inhibitors, and the cell cycle played an important role in the apoptosis induced by PKC inhibitors. .