不同浓度罗哌卡因对肺癌细胞生长和迁移能力的影响

Effects of different concentrations of ropivacaine on growth and migration of lung cancer cells

目的评价不同浓度罗哌卡因对肺癌细胞生长和迁移能力的影响。方法人肺腺癌细胞株A549细胞和人肺鳞癌细胞株H520细胞, 采用随机数字表法分别分成4组(n=24):对照组(C组)和不同浓度罗哌卡因组(Ⅰ~Ⅲ组)。C组正常培养, Ⅰ~Ⅲ组分别加入罗哌卡因3、5和7 mmol/L进行处理。分别于处理24 h(T1)、48 h(T2)和72 h(T3)时, 采用CCK-8法测定细胞存活率;于T1时采用流式细胞术检测细胞周期和细胞凋亡情况;采用Western blot法检测细胞周期素D1(Cyclin D1)、细胞周期调节蛋白依赖性激酶4(CDK4)、cleaved多聚腺苷二磷酸核糖聚合酶1(PARP-1)和cleaved caspase-3的表达;采用划痕实验检测细胞迁移距离;分光光度法检测RhoA和Rac1活性。结果 C组、Ⅰ组、Ⅱ组和Ⅲ组T1~3时A549和H520细胞存活率依次降低, G0/G1期比例和凋亡比例依次增加, T1时Cyclin D1和CDK4表达依次下调, cleaved PARP-1和cleaved caspase-3表达依次上调, 细胞迁移距离、RhoA和Rac1活性依次降低(P<0.05)。结论罗哌卡因可浓度依赖性地抑制肺癌细胞生长和迁移能力, 与诱导细胞周期阻滞和凋亡有关。

Objective To evaluate the effects of different concentrations of ropivacaine on the growth and migration of lung cancer cells.Methods Human lung adenocarcinoma cell strain A549 cells and human lung squamous cell strain H520 cells were divided into 4 groups(n=24 each)using a random number table method:control group(group C)and different concentrations of ropivacaine groups(Ⅰ-Ⅲgroups).Cells were commonly cultured in group C.Ropivacaine 3,5 and 7 mmol/L were added and then the cells were cultured inⅠ-Ⅲgroups,respectively.The cell survival rate was determined using the CCK-8 method at 24,48 and 72 h of treatment(T1-3).The cell cycle and apoptosis were detected at T1 using flow cytometry.The expression of Cyclin D1,cyclin-dependent kinase 4(CDK4),cleaved poly(ADP-ribose)polymerase-1(PARP-1)and cleaved caspase-3 was detected using Western blot.Wound healing assay was used to measure cell migration distance.The activities of RhoA and Rac1 were detected by microplate spectrophotometry.Results The cell viability of A549 and H520 cells sequentially decreased at T1-3,the proportion of G0/G1 phase and apoptosis sequentially increased,the expression of Cyclin D1 and CDK4 was down-regulated sequentially at T1,the expression of cleaved PARP-1 and cleaved caspase-3 was up-regulated sequentially,and the cell migration distance,RhoA,and Rac1 activity decreased sequentially in C,Ⅰ,ⅡandⅢgroups(P<0.05).Conclusions Ropivacaine can inhibit the growth and migration ability of lung cancer cells in a concentration-dependent manner,which is related to induction of cell cycle arrest and apoptosis.