The drug resistance mechanism based on HOTAIR of Yifei Jiedu formula regulating apoptosis and autophagy reversal in lung adenocarcinoma

LIU Lingling; SUN Xiyuan; JIANG Mei; CHEN Molan; HAN Dong

doi:10.3969/j.issn.1009-0460.2025.06.001

Abstract：Objective To investigate the effect of Yifei Jiedu formula combined with cisplatin on the drug resistance of A549/DDP cells after silencing the expression of HOTAIR,and to analyze its possible mechanism.Methods Real-time fluorescence quantitative PCR(qPCR)was used to verify the efficiency of siRNA1,siRNA2 and siRNA3 in interfering with the expression of HOTAIR in A549/DDP cells.The proliferation activity of A549/DDP cells in the cisplatin group,the si-HOTAIR+cisplatin group and the si-HOTAIR+Yifei Jiedu formula(YFJDF)+cisplatin group was detected by the CCK-8 method,and the proliferation inhibition rate and half maximal inhibitory concentration(IC50)were calculated.A549/DDP cells were divided into the control group(NC group),the blank serum group(CS group),the YFJDF group,the si-NC group,the si-HOTAIR group,the si-NC+YFJDF group and the si-HOTAIR+YFJDF group.The expressions of apoptosis-related proteins Bax and Bcl-2,as well as autophagy markers LC3,Beclin1 and P62 proteins in cells were detected by Western blotting.The mRNA expressions of LC3,Beclin1 and P62 in each group were detected by qPCR.The apoptosis rate of cells was detected by Annexin V-FITC/PI double staining flow cytometry.Results The result showed that siRNA2 had the highest efficiency in interfering with the expression of HOTAIR,and its relative expression level decreased to 0.42±0.01.HOTAIR silencing significantly reduced the IC50 value of A549/DDP cells to cisplatin(22.162 g/mL vs.45.091 g/mL),and the Yifei Jiedu formula combined with cisplatin further enhanced cisplatin sensitivity(IC50=13.473 g/mL).Compared with the YFJDF group and the si-HOTAIR group,the Bax protein level in the si-HOTAIR+YFJDF group was significantly upregulated(P<0.001),and the expression of Bcl-2 protein was significantly downregulated(P<0.05).Compared with(2.62±0.49)%in the si-NC group,the apoptosis rate in the si-HOTAIR group significantly increased[(33.03±0.69)%],and the difference was statistically significant(P<0.001).Moreover,the apoptosis rate of the si-HOTAIR+YFJDF group[(45.45±1.4)%]was higher than that of the si-HOTAIR group(P<0.001);The expression levels of LC3 and Beclin1 proteins and mRNA in both the DDP+YFJDF group and the si-HOTAIR+YFJDF group were significantly increased(P<0.05),while the expression of P62 decreased(P<0.05).Conclusion Silenting the expression of HOTAIR can enhance the sensitivity of A549/DDP cells to cisplatin.The Yifei Jiedu Formula effectively reverse the drug resistance of A549/DDP cells by regulating the autophagy pathway mediated by HOTAIR and promoting the apoptosis of A549/DDP cells.