隐丹参酮对顺铂耐药肺癌细胞磷酸戊糖途径相关蛋白的影响及机制探讨

Effect of cryptotanshinone on pentose phosphate pathway related proteins in cisplatin-resistant lung cancer cells and its mechanism

目的考察隐丹参酮(cryptotanshinone,CTS)对顺铂处理顺铂耐药肺癌细胞的存活率及磷酸戊糖途径(pentose phosphate pathway,PPP)相关蛋白表达的影响,并探讨其相关机制。方法体外培养人肺癌A549细胞和顺铂耐药A549/CDDP细胞,qPCR和Western blot分别检测两种细胞磷酸戊糖途径相关的G6PD、PGD、TKT和TALDO1的mRNA及蛋白表达水平的差异;CCK-8法检测隐丹参酮对顺铂处理A549/CDDP细胞后细胞存活率的影响;Western blot检测给予不同浓度隐丹参酮处理后A549/CDDP细胞磷酸戊糖途径相关蛋白G6PD、PGD、TKT和TALDO1以及AKT、NRF2蛋白表达水平的变化。结果与A549细胞相比,A549/CDDP细胞G6PD、PGD、TKT和TALDO1的表达水平均增高;隐丹参酮能够有效降低顺铂处理A549/CDDP细胞的细胞存活率;隐丹参酮作用于A549/CDDP细胞后,磷酸戊糖途径相关蛋白TKT表达水平明显降低,AKT和NRF2蛋白表达水平降低,且这些作用呈现剂量依赖性。结论隐丹参酮能有效增加顺铂耐药A549/CDDP细胞对顺铂的敏感性,其机制可能与抑制AKT、NRF2从而抑制磷酸戊糖途径相关蛋白表达相关。

Objective To investigate the effect of cryptotanshinone(CTS)on cell viability and pentose phosphate pathway(PPP)-related protein expression in cisplatin-resistant lung cancer cells with cisplatin treatment,and to explore its relevant mechanism.Methods The mRNA and protein expression levels of G6PD,PGD,TKT and TALDO1 of the pentose phosphate pathway(PPP)in human lung cancer A549 cells and cisplatin-resistant A549/CDDP cells were measured by qPCR and western blot,respectively.The effect of CTS on cell viability of A549/CDDP cells treated with cisplatin was determined by CCK-8 assay.The protein levels of PPP-related G6PD,PGD TKT,TALDO1 and AKT,NRF2 in A549/CDDP cells treated with different concentrations of CTS were measured by western blot.Results Expression of G6PD,PGD,TKT and TALDO1 in A549/CDDP cells were increased compared with A549 cells.CTS could effectively reduce cell viability of A549/CDDP with cisplatin treatment.CTS can significantly reduce the expression of PPP-related protein TKT,and also decrease the expression of AKT and NRF2 proteins in a dose-dependent manner in A549/CDDP cells.Conclusion CTS can effectively increase the sensitivity of cisplatin-resistant A549/CDDP cells to cisplatin,and the mechanism may be related to the inhibition of AKT and NRF2,thereby inhibiting the expression of PPP-related protein.