【摘要】 目的:研究细胞周期蛋白B1反义全长cDNA(AS-CLB1)对抗Lewis肺癌细胞(LL/2)体内外增殖作用的影响,为将AS-CLB1联合用于人肺癌等恶性肿瘤的治疗提供实验依据。方法:(0.01nmol/L～0.1μmol/L)处理LL/2亲本(LP),LL/2空载体(LV)及LL/2/AS-CLB1(LA)三种细胞,分别于1h及24h后用MTT比色法评估对各组细胞的杀伤作用。将上述三种细胞分别接种C57BL/6小鼠,成瘤后用(15mg/kg.d)处理各组动物,3d1次,一共4次,进行成瘤性及动物生存时间观察;流式细胞仪检测各组动物肿瘤组织细胞周期及凋亡。结果:无论是泰素帝处理1h还是24h,LA细胞存活率较LP、LV两对照均明显降低(P<0.05);其中处理1h后,泰素帝80nmol/L组LA细胞存活率低于对照9倍以上,处理24h后,20nmol/L组LA细胞存活率低于对照7倍左右。LA组小鼠肿瘤体积明显小于对照(P<0.05),接种后30天,LP组肿瘤体积大小为1981.29±318.56mm3,LV组为1673.36±297.96mm3,LA组仅为421.68±30.16mm3。LA组肿瘤组织中细胞在G1期所占比例为(89.7±0.5)%,凋亡率为(79.5±1.2)%,均较对照明显增加(P<0.05)。接种后60天,LA组尚有70%的小鼠存活,LP组仅30%,LV组仅40%存活,LA组动物生存时间明显延长(P<0.01)。结论:AS-CLB1可以明显增强抗LL/2细胞在体内外的增殖作用,此增强效应可能与AS-CLB1诱导细胞发生G1期阻滞及凋亡,增强了抗肿瘤作用有关。更多还原

【Abstract】 Objective: To study the effect of full-length cyclin B1 antisense cDNA(AS-CLB1) on Taxotere in inhibiting proliferation of Lewis lung carcinoma cells(LL/2) in vitro and in vivo and to provide the foundation for the feasibility of treating human lung cancer and other malignancies with AS-CLB1 combined with Taxotere. Methods: LL/2 parent cells, LL/2/vector and LL/2/AS-CLB1 transfectants(LP, LV and LA cells) were treated with Taxotere(0.01 nmol/L~0.1 μmol/L) in vitro. The cytotoxicity of Taxotere was evaluated by MTT assay at 1h and 24h after treatment. After inoculation of the three types of cells, C57BL/6 mice were treated with Taxotere(15 mg/kg/day) once every three days for four cycles. Cell cycle and apoptosis in the tumor tissues were determined by flow cytometry. Results: The survival rate of LA cells treated with taxotere was lower than that of the controls(LP and LV cells; P<0.05). At 1h after treatment with taxotere(80nmol/L), the survival rate was (4.56±1.21)% in LA cells, (46.26±3.53)% in LP cells and (44.25±2.21)% in LV cells. At 24h after treatment with 20nmol/L Taxotere, the survival rate was (5.21±1.39)% in LA cells, whereas it was(36.17±3.63)% in LP and (35.15± 3.63)% in LV cells. The tumor volume in the LA group was also lower than that of the controls(LP and LV group; P<0.05). At 30 days after the inoculation of tumor cells, the tumor volume was 421.68±30.16mm3 in the LA group, 1981.29±318.56mm3 in the LP group and 1673.36±297.96mm3 in the LV group. Compared with the control groups, the cells in tumor tissues of the LA group showed G1 arrest and increased apoptosis(P<0.05). There were (89.7±0.5)% cells in G1 phase and the apoptotic ratio was (79.5±1.2)% in the LA group. Moreover, 70% of the mice in the LA group were still alive at the end of the observation period for the survival studies. In contrast, only 30% of the mice in the LP group and 40% of the mice in the LV group were still alive. The survival rate in the LA group was significantly higher than in the control groups(P<0.01). Conclusion: AS-CLB1 can increase the inhibitory effect of Taxotere on the proliferation of LL/2 cells in vitro and in vivo. It enhances the anti-tumor activity of Taxotere by inducing cell apoptosis and G1 arrest.更多还原