罗哌卡因对结肠癌细胞增殖和裸鼠人结肠癌皮下瘤生长的影响

目的:探究罗哌卡因对结肠癌细胞增殖及肿瘤生长的影响。方法:罗哌卡因处理细胞,CCK8 检测细胞活性,确定用药浓度;肿瘤成球实验检测细胞增殖;流式检测细胞凋亡及细胞周期;Western blot 检测细胞增殖、凋亡标记蛋白及细胞周期标记蛋白的表达。复制结肠癌Xenograft 模型,统计肿瘤生长情况及裸鼠存活率,Tunel 法检测细胞凋亡情况。结果:根据CCK8 实验结果确定罗哌卡因给药浓度分别为20、50、100 μg/ ml。罗哌卡因能显著抑制癌细胞集落形成(17.80±0.51,P<0.001) 和增殖标记蛋白Ki67 (0.32±0.68,P<0.01) 、PCNA(0.14±0.24,P<0.01) 的表达,并体现量效关系。罗哌卡因还可明显促进癌细胞凋亡(12.80±1.24,P<0.01) 和凋亡标记蛋白caspase-3(1.76±1.43,P<0.001)、caspase-9 (1.61±1.26,P<0.001) 的表达。同时,罗哌卡因能诱导细胞发生G2 期阻(40.5%,P<0.01) ,促进周期标记蛋白p53 的表达(1.16±0.65,P<0.01) ,抑制Cyclin A 的表达(0.12±0.12,P<0.05) 。此外,罗哌卡因可显著抑制肿瘤生长(1 247.60±1.37,P<0.01) ,提高结肠癌裸鼠的生存率及诱导肿瘤组织细胞凋亡(78.00±1.45,P<0.001) ,且具有量效关系。结论:罗哌卡因能抑制结肠癌细胞增殖和结肠癌皮下瘤的生长。

Effects of ropivacaine on cell proliferation and tumor growth in colon cancer

Objective: To evaluate the effects of ropivacaine on cell proliferation and tumor growth of colon carcinoma.Methods: CCK8 assay was performed for cell viability.The colony formation assay was performed for cell proliferation.Cell flow apoptosis and cell cycle were measured by Flow cytometry.Protein levels were calculated by Western blot.Meanwhile,nude mice were inoculated with SW480 colon cells and treated with ropivacaine.Tumor volume and survival rate were examined after treatment.Results: The results of CCK8 showed that the optimum concentrations of ropivacaine were 20, 50 and 100 μg/ ml respectively. Ropivacaine dose-dependently inhibited colony formation (17.80±0.51,P<0.001) and expressions of Ki67 (0.32±0.68,P<0.01) and PCNA(0.14±0.24,P<0.01).Meanwhile,treatment with ropivacaine markedly increased apoptosis (12.80±1.24,P<0.01) and protein levels of caspase-3(1.76±1.43,P <0.001) and caspase-9 (1.61±1.26,P <0.001).In addition,ropivacaine notably induced cell cycle arrest (40.5%,P<0.01),expression of p53 (1.16±0.65,P<0.01),and down-regulated the expression level of Cyclin A (0.12±0.12,P <0.05).Furthermore,ropivacaine inhibited tumor growth (1 247.60±1.37,P<0.01),up-regulated survival rate of mice and induced apoptosis of tumor tissue (78.00 ±1.45,P <0.001) in a dose-depended manner.Conclusion:Ropivacaine inhibits cell proliferation and tumor growth of colon cancer.