甲基莲心碱调节丝裂原活化蛋白激酶/c-Jun氨基末端激酶信号通路对宫颈癌细胞恶性生物学行为的影响  

Effect of neferine on malignant biological behaviors of cervical cancer cells by regulating the MAPK/JNK signaling pathway

作　　者：汪振华 朱莉莉 肖苏 彭超 WANG Zhenhua;ZHU Lili;XIAO Su;PENG Chao(Department of Pharmacy,Jingzhou Hospital of Traditional Chinese Medicine,Jingzhou,Hubei 434000;First Department of Gynecology,Jingzhou Hospital of Traditional Chinese Medicine,Jingzhou,Hubei 434000)

机构地区：[1]湖北省荆州市中医医院药学部,湖北荆州434000 [2]湖北省荆州市中医医院妇科一病区,湖北荆州434000

出　　处：《河北中医》2025年第3期450-455,共6页Hebei Journal of Traditional Chinese Medicine

摘　　要：目的观察甲基莲心碱(Nef)通过调节丝裂原活化蛋白激酶(MAPK)/c-Jun氨基末端激酶(JNK)信号通路对宫颈癌细胞恶性生物学行为的影响。方法使用不同浓度的Nef(0、5、10、15、20、25、30μmol/L)干预HeLa细胞,CCK-8法检测细胞存活率。将HeLa细胞分为对照组、Nef低剂量组(5μmol/L Nef)、Nef中剂量组(10μmol/L Nef)、Nef高剂量组(15μmol/L Nef)、SP600125组(15μmol/L Nef+10μmol/L MAPK/JNK信号通路抑制剂SP600125),各组细胞使用相应剂量的药物干预24 h。细胞划痕实验检测各组细胞迁移情况;Transwell小室法检测细胞侵袭;DCFH-DA法检测细胞活性氧(ROS)水平;试剂盒法检测细胞丙二醛(MDA)、总超氧化物歧化酶(SOD)水平;流式细胞术检测细胞凋亡水平;Western blot法检测细胞p-JNK1/2/JNK1/2、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、基质金属蛋白酶2(MMP-2)蛋白表达。结果与0μmol/L相比,5、10、15、20、25、30μmol/L的Nef呈剂量依赖性降低HeLa细胞存活率(P<0.05),半数抑制浓度(IC50值)为(16.96±2.53)μmol/L。为保证Nef对HeLa细胞的作用效果并保证HeLa细胞具有一定存活率方便收集细胞,选取5、10、15μmol/L的Nef浓度作为低、中、高剂量组。与对照组相比,Nef低、中、高剂量组细胞划痕愈合率、细胞侵袭个数、细胞SOD水平、MMP-2、Bcl-2蛋白表达明显降低(P<0.05),细胞ROS、MDA水平、细胞凋亡率以及细胞p-JNK1/2/JNK1/2、Bax蛋白表达明显升高(P<0.05),且呈剂量依赖(P<0.05)。与Nef高剂量组相比,SP600125组细胞划痕愈合率、细胞侵袭个数、细胞SOD水平以及MMP-2、Bcl-2蛋白表达明显升高(P<0.05),细胞ROS、MDA水平、细胞凋亡率以及细胞p-JNK1/2/JNK1/2、Bax蛋白表达明显降低(P<0.05)。结论Nef可通过激活MAPK/JNK信号通路抑制宫颈癌细胞恶性生物学行为。Objective To investigate the effect of neferine(Nef)on the malignant biological behaviors of cervical cancer cells by regulating the mitogen activated protein kinase(MAPK)/c-Jum amino terminal kinase(JNK)signaling pathway.Methods HeLa cells were induced with Nef at various concentrations(0,5,10,15,20,25,30μmol/L),and the cell survival rate was measured by cell counting kit-8(CCK-8)assay.HeLa cells were divided into control group,low-dose Nef group(5μmol/L Nef),medium-dose Nef group(10μmol/L Nef),high-dose Nef group(15μmol/L Nef),and SP600125 group(15μmol/L Nef+10μmol/L MAPK/JNK signal pathway inhibitor SP600125).Cells in each group were intervened with corresponding doses of drugs for 24 h.Cell migration and invasion were detected by cell scratch test and Transwell assay,respectively.Reactive oxygen species(ROS)level was detected by DCFH-DA staining.Malondialdehyde(MDA)and superoxide dismutase(SOD)levels were detected by commercial kits.Apoptosis was detected by flow cytometry.Western blot was used to detect the protein expressions of JNK1/2,p-JNK1/2,B-cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and matrix metalloproteinase-2(MMP-2).Results Compared with blank control,Nef induction at 5,10,15,20,25,and 30μmol/L dose-dependently decreased the survival rate of HeLa cells(P<0.05),with the half-maximal drug inhibitory concentration(IC50)value of(16.96±2.53)μmol/L.In order to ensure the effect of Nef on HeLa cells and a certain survival rate to facilitate the collection of cells,5,10 and 15μmol/L Nef concentrations were selected as low-dose,medium-dose and high-dose groups,respectively.Compared with the control group,the cell scratch healing rate,the number of cell invasions,SOD,and protein levels of MMP-2,and Bcl-2 in the low-dose,medium-dose,and high-dose Nef groups were significantly reduced(P<0.05),while the ROS and MDA levels,apoptosis rate and protein expressions of p-JNK1/2/JNK1/2,and Bax proteins were significantly elevated(P<0.05),both in a dose-dependent manner(P<0.05).Compared with the h

关 键 词：宫颈肿瘤 甲基莲心碱 丝裂原活化蛋白激酶 JNK丝裂原活化蛋白激酶类 肿瘤浸润 肿瘤转移 细胞凋亡 

分 类 号：R737.33[医药卫生—肿瘤] R730.52[医药卫生—临床医学] R73-37R286.91