
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
李贺, 牟琪瑞, 王哲, 王莹, 于丹, 高原
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
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 intervention against cisplatin resistance in A549/DDP cells by regulating the Wnt/β-catenin signaling pathway. Methods The expression of β-catenin was silenced by siRNA interference,and Western blot was used to measure the relative protein expression of β-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. After β-catenin interference,Western blot was used to measure the protein expression levels of β-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% on β-catenin expression(P=0.000). After β-catenin was silenced by RNAi technique,compared with the normal serum group(1.00),the Buzhong Yiqi Decoction+cisplatin group had significantly greater reductions in the protein expression of Survivin(0.247±0.015) and β-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 synergistic effect on inhibiting the Wnt/β-catenin signaling pathway and improving cisplatin resistance in lung adenocarcinoma,and Buzhong Yiqi Decoction has a siβ-catenin transient transfection-like effect.
β-catenin / Survivin / RNAi / 肺癌耐药 / 补中益气汤
β-catenin / Survivin / RNAi / lung cancer drug resistance / Buzhong Yiqi Decoction
R285.5 / R734.2
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