补中益气汤协同Siβ-catenin调控Wnt/β-catenin信号通路干预肺腺癌顺铂耐药性的分子机制研究  

Molecular mechanism of Buzhong Yiqi Decoction in coordination with Siβ-catenin in intervention against cisplatin resistance in lung adenocarcinoma by regulating the Wnt/β-catenin signaling pathway

作　　者：李贺[1] 牟琪瑞 王哲[1] 王莹[1] 于丹[1] 高原[1] Li He;Mu Qirui;Wang Zhe;Wang Ying;Yu Dan;Gao Yuan(Department of Pathology,College of Integrative Medicine,Liaoning University of Traditional Chinese Medicine)

机构地区：[1]辽宁中医药大学中西医结合学院病理教研室,沈阳110847

出　　处：《重庆医科大学学报》2024年第2期147-152,共6页Journal of Chongqing Medical University

基　　金：辽宁省教育厅科学研究面上资助项目(编号:LJKZ0901)。

摘　　要：目的:探讨补中益气汤含药血清协同Siβ-catenin调控Wnt/β-catenin信号通路干预A549/DDP细胞顺铂耐药性的分子机制研究。方法:采用siRNA干扰沉默β-catenin的表达,Western blot检测正常血清组、顺铂组、siCTNNB1-235组、siCTNNB1-510组、siCTNNB1-547组A549/DDP细胞的β-catenin蛋白的相对表达量;干扰β-catenin后,Western blot检测各组A549/DDP细胞β-catenin和Survivin的蛋白表达量,MTT法检测各组A549/DDP细胞对顺铂的IC50,Annexin V/PI染色法检测各组A549/DDP细胞顺铂诱导的凋亡率。结果:相较于正常血清组(1.00),siCTNNB1-235组(0.323±0.021)对β-catenin表达抑制率达67%(P=0.000)。RNAi技术沉默β-catenin后,与正常血清组(1.00)相比,补中益气汤联合顺铂组的Survivin(0.247±0.015)和β-catenin(0.257±0.015)蛋白的表达下调更为明显(P=0.000,P=0.000)。IC50和正常血清组(28.330±1.029)µmol/L相比,单独补中益气汤含药血清(20.350±1.155)µmol/L或单独瞬时转染siCTNNB1-235组(15.577±1.535)µmol/L均可降低A549/DDP细胞顺铂的IC50(P=0.000,P=0.000),2者联合(10.453±0.999)µmol/L使用可进一步降低顺铂的IC50(P=0.000)。与正常血清组(9.130±0.384)%相比,瞬时转染siCTNNB1-235联合顺铂组(64.393±0.244)%和补中益气汤联合顺铂组(70.120±0.400)%的总凋亡率均升高(P=0.000,P=0.000)。结论:补中益气汤含药血清和瞬时转染siβ-catenin在抑制Wnt/β-catenin信号通路改善肺腺癌顺铂耐药性方面具有协同效应,且补中益气汤具有siβ-catenin瞬时转染样效应。Objective:To investigate the molecular mechanism of Buzhong Yiqi Decoction in coordination with Siβ-catenin in inter⁃vention against cisplatin resistance in A549/DDP cells by regulating the Wnt/β-catenin signaling pathway.Methods:The expression ofβ-catenin was silenced by siRNA interference,and Western blot was used to measure the relative protein expression ofβ-catenin in A549/DDP cells in the normal serum group,the cisplatin group,the siCTNNB1-235 group,the siCTNNB1-510 group,and the siCTNNB1-547 group.Afterβ-catenin interference,Western blot was used to measure the protein expression levels ofβ-catenin and Survivin in A549/DDP cells of each group;MTT assay was used to determine the IC50 of cisplatin in A549/DDP cells of each group;Annexin V/PI staining was used to measure the cisplatin-induced apoptosis rate of A549/DDP cells in each group.Results:Compared with the normal serum group(1.00),the siCTNBN1-235 group(0.323±0.021)had an inhibition rate of 67%onβ-catenin expression(P=0.000).Afterβ-catenin was silenced by RNAi technique,compared with the normal serum group(1.00),the Buzhong Yiqi Decoc⁃tion+cisplatin group had significantly greater reductions in the protein expression of Survivin(0.247±0.015)andβ-catenin(0.257±0.015)(P=0.000,P=0.000).As for the IC50 of cisplatin in A549/DDP cells,compared with the normal serum group(28.330±1.029)µmol/L,serum containing Buzhong Yiqi decoction alone(20.350±1.155)µmol/L or siCTNNB1-235 transient transfection alone(15.577±1.535)µmol/L reduced the IC50 of cisplatin in A549/DDP cells(P=0.000,P=0.000),and the combination of the two methods(10.453±0.999)µmol/L further reduced the IC50 of cisplatin(P=0.000).Compared with the normal serum group(9.130±0.384)%,the siCTNBN1-235 transient transfection+cisplatin group(64.393±0.244)%and the Buzhong Yiqi Decoction+cisplatin group(70.120±0.400)%had a significant increase in overall apoptosis rate(P=0.000,P=0.000).Conclusion:Serum containing Buzhong Yiqi Decoction and siβ-catenin transient transfection have a

关 键 词：Β-CATENIN Survivin RNAI 肺癌耐药 补中益气汤 

分 类 号：R285.5[医药卫生—中药学] R734.2[医药卫生—中医学]