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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.
To investigate the effect of antisense c-fos and c-jun on the growth of CNE-2Z cells and its relationship with the expression of PKC-α, antisense c-fos and c-jun oligonucleotides were used in combination with CNE-2Z The cell growth index was determined by MTT assay and the expression of PKC-α was detected by flow cytometry (FCM) .The results showed that the minimum effective concentration of c-fos and c-jun antisense oligonucleotides was 1.7 × The inhibitory effect on CNE-2Z cells was enhanced with the concentration of 10 -6 μg / ml, the minimum effective concentration was 1.7 × 10 -5 μg / ml. After a certain period of time, (P <0.01), and the maximum inhibitory effect time was 20 hours and the growth index was 63.9 ± 5.0, which was significantly lower than that of the control group (P <0.01) .In the above concentration of LP and antisense oligonucleotide After 20 hours, FCM assay showed that the fluorescence intensity of PKC-α was significantly lower than that of each control group, the percentage of positive cells was 10.40 ± 5.23, which was significantly lower than that of control group (29.87 ± 7.41) and other groups (P <0.01). The results showed that antisense c-fos and c-jun oligonucleotides could inhibit the growth and proliferation of CNE-2Z cells in a dose- and time-dependent manner and significantly inhibited the expression of PKC-α protein expression.