罗哌卡因对食管鳞癌细胞FAK/ERK通路及细胞学行为的影响

Effects of Ropivacaine on FAK/ERK Pathway and Cytological Behavior of Esophageal Squamous Cell Carcinoma

目的:观察罗哌卡因对人食管鳞状癌细胞(KYSE150)增殖、迁移、侵袭和凋亡的影响,并探讨其作用机制。方法:体外培养KYSE150细胞,随机分为对照组、罗哌卡因Ⅰ、Ⅱ、Ⅲ组(10,20,50μg·ml-1)。采用噻唑蓝(MTT)法检测罗哌卡因对KYSE150细胞的增殖抑制作用;划痕实验检测各组KYSE150细胞迁移能力变化情况;Transwell法检测各组KYSE150细胞的侵袭能力;Annexin V-FITC/PI双染法检测各组KYSE150细胞凋亡情况;蛋白免疫印迹分析法检测黏着斑激酶(FAK)、磷酸化FAK(p-FAK)、信号调节激酶(ERK)、磷酸化ERK (p-ERK)蛋白表达情况。结果:与对照组相比,罗哌卡因Ⅰ、Ⅱ、Ⅲ组KYSE150细胞存活率依次降低(P<0.05),并呈剂量依赖性(P<0.05);与对照组相比,罗哌卡因Ⅰ组KYSE150细胞愈合率、侵袭数量、凋亡率、p-FAK和p-ERK蛋白表达量差异均无统计学差异(P>0.05);与对照组及罗哌卡因Ⅰ组相比,罗哌卡因Ⅱ、Ⅲ组KYSE150细胞愈合率、侵袭数量、p-FAK和p-ERK蛋白表达量依次降低(P<0.05),KYSE150细胞凋亡率依次升高(P<0.05);各组KYSE150细胞中FAK和ERK蛋白表达量差异无统计学意义(P>0.05)。结论:罗哌卡因可能通过下调p-FAK/pERK蛋白表达水平从而影响人食管鳞状癌细胞迁移、侵袭和凋亡行为。

Objective: To observe the effects of ropivacaine on the proliferation,migration,invasion and apoptosis of human esophageal squamous cell carcinoma(KYSE150),and to explore its mechanism.Methods: The KYSE150 cells were cultured in vitro and randomly divided into control group and ropivacaine groups Ⅰ,Ⅱ and Ⅲ(10,20,50 μg·ml-1).The inhibitory effect of ropivacaine on the proliferation of KYSE150 cells was detected by MTT method;the migration ability of KYSE150 cells was detected by scratch test;the invasion of KYSE150 cells was detected by Transwell method;the apoptosis of KYSE150 cells was detected by Annexin V-FITC/PI double staining;the expressions of focal adhesion kinase(FAK),phosphorylation-FAK(p-FAK),signal-regulated kinase(ERK) and phosphorylation-ERK(p-ERK) protein were detected by Western blot.Results: Compared with that in the control group,the survival rate of KYSE150 cells in ropivacaine groups Ⅰ,Ⅱ and Ⅲ decreased in a dose-dependent manner(P < 0.05);there were no significant differences in the healing rate,invasion number,apoptosis rate and p-FAK and p-ERK protein expressions between ropivacaine group Ⅰ and the control group(P>0.05);compared with those in the control group and ropivacaine group Ⅰ,the healing rate of KYSE150 cells,invasion number,p-FAK and p-ERK protein expressions in ropivacaine groups Ⅱ and Ⅲ decreased in turn(P<0.05),and the apoptosis rate of KYSE150 cells increased in turn(P<0.05);there were no significant differences in the expressions of FAK and ERK protein in KYSE150 cells of each group(P>0.05).Conclusion: Ropivacaine may affect the migration,invasion and apoptosis of human esophageal squamous cell carcinoma by down-regulating the expression of p-FAK/p-ERK protein.