山甲白花汤联合吉西他滨调控miR-124-3p表达逆转胰腺癌耐药的实验研究  

Experimental study of Shanjia Baihua Decoction combined with gemcitabine to regulate miR-124-3p expression and reverse drug resistance in pancreatic cancer

作　　者：孙素芹[1] 高磊[2] 叶婷[3] 樊蓉[4] 刘春婷 刘冬影 常丽 SUN Suqin;GAO Lei;YE Ting(Department of Pulmonology,The Second Affiliated Hospital of Heilongjiang University of TCM,Harbin 150001,China;不详)

机构地区：[1]黑龙江中医药大学附属第二医院肺病科,哈尔滨150001 [2]黑龙江中医药大学附属第二医院肿瘤科,哈尔滨150001 [3]黑龙江中医药大学附属第二医院内分泌科,哈尔滨150001 [4]黑龙江中医药大学附属第二医院哈南分院内分泌科,哈尔滨150060 [5]黑龙江中医药大学附属第二医院哈南分院肺病科,哈尔滨150060 [6]哈尔滨市胸科医院肿瘤科,哈尔滨150056

出　　处：《中国处方药》2024年第12期48-52,共5页Journal of China Prescription Drug

基　　金：黑龙江省卫生健康委科研课题(20212121020349)。

摘　　要：目的探究山甲白花汤联合吉西他滨(GEM)调控miR-124-3p表达逆转胰腺癌耐药的机制。方法qRT-PCR检测miR-124-3p表达;细胞计数试剂盒(CCK-8)检测细胞对GEM敏感性(IC_(50));Western blot检测Bcl-2、MDR1、MRP1、Caspase-3、β-catenin、c-Myc、Cyclin D1蛋白表达;EdU实验检测细胞增殖率;Transwell检测细胞迁移数。结果与PANC-1-GEM+吉西他滨组相比,PANC-1-GEM+联合组上调了miR-124-3p表达[(0.75±0.06)vs.(1.20±0.11)],降低了PANC-1-GEM细胞IC_(50)[(26.34±3.42)μM vs.(19.52±2.86)μM]、耐药蛋白MDR1[(3.08±0.11)vs.(1.75±0.10)]及MRP1[(3.14±0.10)vs.(1.46±0.10)]的表达,上调凋亡蛋白Bcl-2[(1.75±0.12)vs.(3.18±0.14)]表达并下调Caspase-3的表达[(0.87±0.08)vs.(0.41±0.06)],抑制了细胞增殖率[(65.45±5.44)%vs.(42.69±4.18)%]及细胞迁移数[(66.59±7.81)个vs.(47.20±5.36)个],同时抑制了β-catenin[(0.78±0.10)vs.(0.52±0.07)]、c-Myc[(0.85±0.09)vs.(0.62±0.08)]、Cyclin D1[(0.60±0.08)vs.(0.33±0.07)]蛋白的表达。与PANC-1-GEM+联合+NC inhibitor组相比,PANC-1-GEM+联合+miR-124-3p inhibitor组抑制了miR-124-3p表达[(1.19±0.10)vs.(0.35±0.06)],上调了PANC-1-GEM细胞IC_(50)[(20.68±3.07)μM vs.(31.08±3.45)μM]、耐药蛋白MDR1[(1.73±0.12)vs.(4.02±1.13)]及MRP1[(1.47±0.11)vs.(3.68±0.14)]的表达,抑制凋亡蛋白Bcl-2[(3.07±0.12)vs.(1.71±0.10)]表达并促进Caspase-3的表达[(0.42±0.08)vs.(0.94±0.10)],促进细胞增殖率[(43.10±4.05)%vs.(74.38±5.20)%]及细胞迁移数[(48.35±5.28)个vs.(86.79±6.27)个],同时上调β-catenin[(0.53±0.09)vs.(1.05±0.11)]、c-Myc[(0.63±0.10)vs.(1.20±0.13)]、Cyclin D1[(0.32±0.08)vs.(0.96±0.12)]蛋白的表达。结论山甲白花汤联合吉西他滨通过上调miR-124-3p发挥抗胰腺癌耐药作用,该机制可能通过Wnt/β-catenin通路。Objective To explore whether Shanjia Baihua Decoction combined with gemcitabine(GEM)can reverse drug resistance of pancreatic cancer by regulating the expression of miR-124-3p.Methods The expression of miR-124-3p was detected by qRT-PCR.Cell counting kit(CCK-8)was used to detect GEM sensitivity(IC_(50)).Western blotwas detected Bcl-2,MDR1,MRP1,Caspase-3,β-catenin,c-Myc,Cyclin D1 protein expression.The cell proliferation rate was detected by EdU assay.Transwell detected the number of cell migrations.Results Compared with the PANC-1-GEM+gemcitabine group,the PANC-1-GEM+combination group up-regulated miR-124-3p expression[(0.75±0.06)vs.(1.20±0.11)]and decreased the IC_(50) of PANC-1-GEM cells[(26.34±3.42)μM vs.(19.52±2.86)μM],expression of resistance proteins MDR1[(3.08±0.11)vs.(1.75±0.10)]and MRP1[(3.14±0.10)vs.(1.46±0.10)],up-regulated the expression of apoptotic protein Bcl-2[(1.75±0.12)vs.(3.18±0.14)]and down-regulated the expression of Caspase-3[(0.87±0.08)vs.(0.41±0.06)],inhibited the cell proliferation rate[(65.45±5.44)%vs.(42.69±4.18)%]and the number of cells migrating[(66.59±7.81)vs.(47.20±5.36)],and also inhibitedβ-catenin[(0.78±0.10)vs.(0.52±0.07)],c-Myc[(0.85±0.09)vs.(0.62±0.08)],and Cyclin D1[(0.60±0.08)vs.(0.33±0.07)]proteins expression.Compared with the PANC-1-GEM+combined+NC inhibitor group,the PANC-1-GEM+combined+miR-124-3p inhibitor group suppressed miR-124-3p expression[(1.19±0.10)vs.(0.35±0.06)],up-regulated the IC_(50) of PANC-1-GEM cells[(20.68±3.07)μM vs.(31.08±3.45)μM],expression of drug-resistant proteins MDR1[(1.73±0.12)vs.(4.02±1.13)]and MRP1[(1.47±0.11)vs.(3.68±0.14)],and inhibition of apoptotic protein Bcl-2 expression[(3.07±0.12)vs.(1.71±0.10)]and promoted the expression of Caspase-3[(0.42±0.08)vs.(0.94±0.10)],and promoted the cell proliferation rate[(43.10±4.05)%vs.(74.38±5.20)%]and the number of cells migrated[(48.35±5.28)vs.(86.79±6.27)],while up-regulating the expression ofβ-catenin[(0.53±0.09)vs.(1.05±0.11)],c-Myc[(0.63±0.10)vs.(1.20�

关 键 词：山甲白花汤 吉西他滨 胰腺癌 WNT/Β-CATENIN通路 耐药性 miR-124-3p 

分 类 号：R735.9[医药卫生—肿瘤]