罗哌卡因对胃癌BGC-823细胞生物学行为及AMPK/Nrf2信号通路的影响

Effects of ropivacaine on biological behavior and AMPK/Nrf2 signaling pathway of gastric cancer BGC-823 cells

目的探究罗哌卡因对胃癌BGC-823细胞生物学行为及腺苷酸活化蛋白激酶(AMPK)/核因子E2相关因子2(Nrf2)信号通路的影响。方法体外培养胃癌BGC-823细胞,取培养至对数生长期的BGC-823细胞分为对照组(常规培养)、顺铂组(培养基含5 mg/L顺铂)及罗哌卡因低水平(培养基含25 mg/L罗哌卡因)、中水平(培养基含50 mg/L罗哌卡因)、高水平(培养基含100 mg/L罗哌卡因)组,培养48 h。采用细胞计数试剂-8(CCK-8)检测BGC-823细胞存活率;Transwell小室实验检测BGC-823细胞侵袭能力;流式细胞仪检测BGC-823细胞凋亡率;实时荧光定量PCR(qPCR)法检测BGC-823细胞中AMPK、Nrf2信使RNA(mRNA)水平;Western blot法检测BGC-823细胞中AMPK、Nrf2蛋白水平。结果与对照组比较,顺铂组及罗哌卡因低、中、高水平组BGC-823细胞存活率、侵袭数量、AMPK、Nrf2 mRNA和蛋白水平均下降(P<0.05),而凋亡率升高(P<0.05);与顺铂组比较,罗哌卡因低、中、高水平组BGC-823细胞存活率、侵袭数量、AMPK、Nrf2 mRNA和蛋白水平均升高(P<0.05),而凋亡率下降(P<0.05);与罗哌卡因低水平组比较,罗哌卡因中、高水平组BGC-823细胞存活率、侵袭数量、AMPK、Nrf2 mRNA和蛋白水平均依次下降(P<0.05),而凋亡率依次升高(P<0.05)。结论罗哌卡因能抑制BGC-823细胞存活和侵袭,促进BGC-823细胞凋亡,其机制可能与抑制AMPK/Nrf2信号通路有关。

Objective To investigate the effect of ropivacaine on biological behavior of gastric cancer BGC-823 cells and AMP activated protein kinase(AMPK)/nuclear factor E2-related factor 2(Nrf2)signaling pathway.Methods Gastric cancer BGC-823 cells were cultured in vitro.When BGC-823 cells were cultured to logarithmic growth phase,they were divided into control group(routine culture),cisplatin group(medium containing 5 mg/L cisplatin),and low-level(medium containing 25 mg/L ropivacaine),medium-level(medium containing 50 mg/L ropivacaine),and high-level(medium containing 100 mg/L ropivacaine)ropivacaine groups.The cells were cultured for 48 h.Cell counting kit-8(CCK-8)was used to detect the survival rate of BGC-823 cells.Transwell chamber assay was used to detect the invasion ability of BGC-823 cells.The apoptosis rate of BGC-823 cells was detected by flow cytometry.The mRNA levels of AMPK and Nrf2 in BGC-823 cells were detected by real-time fluorescence quantitative PCR(qPCR).The protein levels of AMPK and Nrf2 in BGC-823 cells were detected by Western blot.Results Compared with the control group,the survival rate,invasion number,AMPK and Nrf2 mRNA and protein levels of BGC-823 cells in the cisplatin group and the low-level,medium-level and high-level ropivacaine groups were decreased(P<0.05),while the apoptosis rate was increased(P<0.05).Compared with the cisplatin group,the survival rate,invasion number,AMPK and Nrf2 mRNA and protein levels of BGC-823 cells in the low-level,medium-level and high-level ropivacaine groups were increased(P<0.05),while the apoptosis rate was decreased(P<0.05).Compared with the low-level ropivacaine group,the survival rate,invasion number,AMPK,Nrf2 mRNA and protein levels of BGC-823 cells in the medium-level and high-level ropivacaine groups were decreased in oder(P<0.05),while the apoptosis rate was increased in oder(P<0.05).Conclusion Ropivacaine could inhibit the survival and invasion of BGC-823 cells and promote the apoptosis of BGC-823 cells,which may be related to the inhibition of AMPK/Nrf2 signaling pathway.