反义c-fos、c-jun对CNE-2Z细胞增殖及PKC-α表达的影响

Inhibitive Effects of Antisense Oligonucleotides of c-fos and c-jun on the Proliferation of the CNE-2Z Cells and the Expression of PKC-α

为探讨反义c-fos、c-jun对CNE-2Z细胞生长的影响及与PKC-α表达的关系.本研究联合应用反义c-fos和c-jun寡核苷酸作用于CNE-2Z细胞,MTT法检测细胞生长指数,流式细胞仪(FCM)检测细胞PKC-α表达.结果显示,c-fos、c-jun反义寡核苷酸在Lipofectin(LP)最低有效浓度为1.7×10^(-6)μg/ml作用下,对CNE-2Z细胞抑制作用随浓度增大而增强,最低有效浓度为1.7×10^(-5)μg/ml;在一定时间内,对细胞生长的抑制作用,随作用时间延长而增强,最大抑制作用时间为20小时,其生长指数为63.9±5.0,较对照组明显降低(P<0.01).在以上浓度的LP和反义寡核苷酸作用于细胞20小时后,FCM检测发现PKC-α荧光强度明显低于各对照组,阳性细胞数百分比为10.40±5.23,较对照组29.87±7.41及其它各组均显著降低(P<0.01).结果表明,反义c-fos、c-jun寡核苷酸可抑制CNE-2Z细胞生长增殖,并呈明显的浓度和时间效应依赖关系,且明显抑制PKC-α蛋白的表达.

The effects of antisense oligonucleotides of c-fos and c-jun on the proliferation of CNE-2Z cells and on the expression of protein kinase C-α (PKC-α) were investigated. With the interpoising of Lipofectin(LP), the CNE-2Z cells were added with antisens oligonucleotides of c-fos and c-jun. MTT method was used to test the cell proliferation and flow cytometry (FCM) to the PKC-a expression. The results showed that the inhibition of antisense oligonucleotides of c-fos and c-jun to CEN-2Z cells was gradually enhanced with its concentration and prolonging time increasing and that its lowest effective concentration of LP was 1.7 X 10^(-6)μg/ml, the lowest effective concentration was of antisense oligonucleotides of c-fos and c-jun 1.7 x 10^(-5)μg/ml, and the most effective time was 20 hours after treatment. The fluoresence of PKC-a and percentage of positive cells in the groups treated by antisense oligonucleotides of c-fos and c-jun decreased significantly. The results indicated that the autisense oligonucleotides of c-fos and c-jun could inhibit the proliferation of CNE-2Z ceUs the expression of PKC-α.