氯硝柳胺联合顺铂抑制肾上腺皮质癌的实验研究
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摘要
目的研究氯硝柳胺和低剂量顺铂联合应用对人肾上腺皮质癌SW-13细胞株增殖、凋亡,以及迁移力、侵袭力的影响,并探讨其作用机制。方法体外培养SW-13细胞,CCK-8法检测氯硝柳胺和不同浓度顺铂对细胞增殖的影响。设对照组、氯硝柳胺组、顺铂组、联合用药组,分别用Annexin V/PI和吖啶橙/溴化乙锭(AO/EB)染色检测细胞凋亡率的变化,Transwell实验和细胞划痕实验检测细胞侵袭、迁移能力的变化,Western blot检测B细胞淋巴瘤因子2(Bcl-2)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、波形蛋白(Vimnetin)、E-钙黏蛋白(E-cadherin)表达的变化。结果氯硝柳胺增强顺铂对SW-13细胞增殖的抑制作用。Annexin V/PI和AO/EB染色结果表明,与联合用药组比较,氯硝柳胺、顺铂促进细胞凋亡(P<0.05)。Transwell和划痕实验结果表明,与联合用药组比较,氯硝柳胺、顺铂降低SW-13细胞的侵袭能力(P<0.05)。Western blot检测结果表明,与联合用药组比较,氯硝柳胺、顺铂抑制Bcl-2、Vimentin的表达,促进Caspase-3、E-cadherin的表达(P<0.05)。结论氯硝柳胺能够增强低剂量顺铂对SW-13细胞增殖的抑制作用,促进细胞凋亡,抑制侵袭及迁移能力,可能与其抑制Bcl-2、Caspase-3、E-cadherin、Vimentin的表达有关。
Objective To study the effect of Niclosamide combined with low-dosage cisplatin on the proliferation,apoptosis,migration and invasion of human adrenocortical carcinoma SW-13 cell line and its potential mechanism.Methods SW-13 cells were cultured in vitro.CCK-8 method was used to detect the influence of Niclosamide and different concentrations of cisplatin on cell proliferation.There were four groups:a control group,a Niclosamide(1μmol/L)group,a cisplatin(16 mg/L)group,and a combined Niclosamide(1μmol/L)plus cisplatin(16 mg/L)group.The change of cell apoptosis was detected with Annexin V/PI staining flow cytometry and AO/EB staining method respectively.The changes of cell invasion and migration abilities were detected by Transwell assay and cell scratch assay.Western blot was used to detect the changes of expressions of Bcl-2,caspase-3,E-cadherin and vimentin.Results Niclosamide enhanced the inhibitory effect of cisplatin on proliferation of SW-13 cells(P<0.05).The results of Annexin V/PI staining and AO/EB staining showed that the combined medicine group enhanced cell apoptosis rate more significantly than the Niclosamide group and the cisplatin group(P<0.05).Transwell assay showed that compared with the Niclosamide and cisplatin groups,the combined medicine group decreased the invasion ability of SW-13 cells(P<0.05),and cell scratch experiments showed that compared with the Niclosamide and cisplatin groups,the combined medicine group decreased the migration ability of SW-13 cells(P<0.05).Western blot showed that compared with the Niclosamide and cisplatin groups,the combined medicine group more efficiently inhibited the expressions of Bcl-2 and vimentin(P<0.05),promoted the expressions of caspase-3 and E-cadherin(P<0.05).Conclusions Niclosamide can enhance the inhibitory effect of low-dosage cisplatin on the proliferation of SW-13 cells,promote cell apoptosis,and inhibit cell invasion and migration abilities.This is probably related to its influence on expressions of Bcl-2,caspase-3,E-cadherin and vimentin.
Objective To study the effect of Niclosamide combined with low-dosage cisplatin on the proliferation,apoptosis,migration and invasion of human adrenocortical carcinoma SW-13 cell line and its potential mechanism.Methods SW-13 cells were cultured in vitro.CCK-8 method was used to detect the influence of Niclosamide and different concentrations of cisplatin on cell proliferation.There were four groups:a control group,a Niclosamide(1μmol/L)group,a cisplatin(16 mg/L)group,and a combined Niclosamide(1μmol/L)plus cisplatin(16 mg/L)group.The change of cell apoptosis was detected with Annexin V/PI staining flow cytometry and AO/EB staining method respectively.The changes of cell invasion and migration abilities were detected by Transwell assay and cell scratch assay.Western blot was used to detect the changes of expressions of Bcl-2,caspase-3,E-cadherin and vimentin.Results Niclosamide enhanced the inhibitory effect of cisplatin on proliferation of SW-13 cells(P<0.05).The results of Annexin V/PI staining and AO/EB staining showed that the combined medicine group enhanced cell apoptosis rate more significantly than the Niclosamide group and the cisplatin group(P<0.05).Transwell assay showed that compared with the Niclosamide and cisplatin groups,the combined medicine group decreased the invasion ability of SW-13 cells(P<0.05),and cell scratch experiments showed that compared with the Niclosamide and cisplatin groups,the combined medicine group decreased the migration ability of SW-13 cells(P<0.05).Western blot showed that compared with the Niclosamide and cisplatin groups,the combined medicine group more efficiently inhibited the expressions of Bcl-2 and vimentin(P<0.05),promoted the expressions of caspase-3 and E-cadherin(P<0.05).Conclusions Niclosamide can enhance the inhibitory effect of low-dosage cisplatin on the proliferation of SW-13 cells,promote cell apoptosis,and inhibit cell invasion and migration abilities.This is probably related to its influence on expressions of Bcl-2,caspase-3,E-cadherin and vimentin.
引文
[1]RUTTER C E,PARK H S,CORSO C D,et al.Addition of radiotherapy to adjuvant chemotherapy is associated with improved overall survival in resected pancreatic adenocarcinoma:an analysis of the national cancer data base[J].Cancer,2015,121(23):4141-4149.
[2]POSTLEWAIT L M,ETHUN C G,TRAN T B,et al.Outcomes of adjuvant mitotane after resection of adrenocortical carcinoma:a13-institution study by the US adrenocortical carcinoma group[J].Journal of the American College of Surgeons,2016,222(4):480-490.
[3]SHEN Z,QIN L,XU T,et al.Chloroquine enhances the efficacy of cisplatin by suppressing autophagy in human adrenocortical carcinoma treatment[J].Drug Design,Development and Therapy,2016,10(5):1035-1045.
[4]LIU J,CHEN X,WARD T,et al.Combined niclosamide with cisplatin inhibits epithelial-mesenchymal transition and tumor growth in cisplatin-resistant triple-negative breast cancer[J].Tumor Biology,2016,37(7):9825-9835.
[5]MARGONIS G A,KIM Y,TRAN T B,et al.Outcomes after resection of cortisol-secreting adrenocortical carcinoma[J].The American Journal of Surgery,2016,211(6):1106-1113.
[6]GHORAYEB N E,RONDEAU G,LATOUR M,et al.Rapid and complete remission of metastatic adrenocortical carcinoma persisting 10 years after treatment with mitotane monotherapy[J].Medicine,2016,95(13):e3180-e3181.
[7]吕雨虹,陈庆,赵娟,等.转染白细胞介素18基因通过下调多药耐药基因表达增强顺铂对C6胶质瘤细胞毒作用[J].中国现代医学杂志,2016,26(10):20-24.
[8]RISBERG K,REDALEN K R,SONSTEVOLD L,et al.Prosurvival responses to the dual inhibition of anti-apoptotic Bcl-2 family proteins and m TOR-mediated signaling in hypoxic colorectal carcinoma cells[J].BMC Cancer,2016,16(1):531-532.
[9]AKHTAR K,ARA A,SIDDIQUI S A,et al.Transition of Immunohistochemical expression of E-cadherin and vimentin from premalignant to malignant lesions of oral cavity and oropharynx[J].Oman Med J,2016,31(3):165-169.
[10]张广伟,朱朝阳,李铁强,等.可溶性E钙蛋白与膀胱尿路上皮癌的相关性[J].中国现代医学杂志,2013,23(17):21-24.
[11]VARGAS D A,SUN M,SADYKOV K,et al.The Integrated role of Wnt/β-catenin,n-glycosylation,and e-cadherin-mediated adhesion in network dynamics[J].PLo S Computational Biology,2016,12(7):e1005007-e1005008.
[12]LIN C K,BAI M Y,HU T M,et al.Preclinical evaluation of a nanoformulated antihelminthic,niclosamide,in ovarian cancer[J].Oncotarget,2016,7(8):8993-9006.
[13]CIRCU M L,DYKES S S,CARROLL J,et al.A novel high content imaging-based screen identifies the anti-helminthic niclosamide as an inhibitor of lysosome anterograde trafficking and prostate cancer cell invasion[J].PLo S One,2016,11(1):e146931-e146932.
[14]TOMIZAWA M,SHINOZAKI F,MOTOYOSHI Y,et al.Niclosamide suppresses migration of hepatocellular carcinoma cells and downregulates matrix metalloproteinase-9 expression[J].Oncol Lett,2015,10(6):3515-3518.
[15]卞震炯,杨冬花.益气温阳活血方对慢性心力衰竭心室重塑患者外周血单个核细胞Bax/Bcl-2表达的影响[J].中国现代医学杂志,2015,25(34):6-9.
[2]POSTLEWAIT L M,ETHUN C G,TRAN T B,et al.Outcomes of adjuvant mitotane after resection of adrenocortical carcinoma:a13-institution study by the US adrenocortical carcinoma group[J].Journal of the American College of Surgeons,2016,222(4):480-490.
[3]SHEN Z,QIN L,XU T,et al.Chloroquine enhances the efficacy of cisplatin by suppressing autophagy in human adrenocortical carcinoma treatment[J].Drug Design,Development and Therapy,2016,10(5):1035-1045.
[4]LIU J,CHEN X,WARD T,et al.Combined niclosamide with cisplatin inhibits epithelial-mesenchymal transition and tumor growth in cisplatin-resistant triple-negative breast cancer[J].Tumor Biology,2016,37(7):9825-9835.
[5]MARGONIS G A,KIM Y,TRAN T B,et al.Outcomes after resection of cortisol-secreting adrenocortical carcinoma[J].The American Journal of Surgery,2016,211(6):1106-1113.
[6]GHORAYEB N E,RONDEAU G,LATOUR M,et al.Rapid and complete remission of metastatic adrenocortical carcinoma persisting 10 years after treatment with mitotane monotherapy[J].Medicine,2016,95(13):e3180-e3181.
[7]吕雨虹,陈庆,赵娟,等.转染白细胞介素18基因通过下调多药耐药基因表达增强顺铂对C6胶质瘤细胞毒作用[J].中国现代医学杂志,2016,26(10):20-24.
[8]RISBERG K,REDALEN K R,SONSTEVOLD L,et al.Prosurvival responses to the dual inhibition of anti-apoptotic Bcl-2 family proteins and m TOR-mediated signaling in hypoxic colorectal carcinoma cells[J].BMC Cancer,2016,16(1):531-532.
[9]AKHTAR K,ARA A,SIDDIQUI S A,et al.Transition of Immunohistochemical expression of E-cadherin and vimentin from premalignant to malignant lesions of oral cavity and oropharynx[J].Oman Med J,2016,31(3):165-169.
[10]张广伟,朱朝阳,李铁强,等.可溶性E钙蛋白与膀胱尿路上皮癌的相关性[J].中国现代医学杂志,2013,23(17):21-24.
[11]VARGAS D A,SUN M,SADYKOV K,et al.The Integrated role of Wnt/β-catenin,n-glycosylation,and e-cadherin-mediated adhesion in network dynamics[J].PLo S Computational Biology,2016,12(7):e1005007-e1005008.
[12]LIN C K,BAI M Y,HU T M,et al.Preclinical evaluation of a nanoformulated antihelminthic,niclosamide,in ovarian cancer[J].Oncotarget,2016,7(8):8993-9006.
[13]CIRCU M L,DYKES S S,CARROLL J,et al.A novel high content imaging-based screen identifies the anti-helminthic niclosamide as an inhibitor of lysosome anterograde trafficking and prostate cancer cell invasion[J].PLo S One,2016,11(1):e146931-e146932.
[14]TOMIZAWA M,SHINOZAKI F,MOTOYOSHI Y,et al.Niclosamide suppresses migration of hepatocellular carcinoma cells and downregulates matrix metalloproteinase-9 expression[J].Oncol Lett,2015,10(6):3515-3518.
[15]卞震炯,杨冬花.益气温阳活血方对慢性心力衰竭心室重塑患者外周血单个核细胞Bax/Bcl-2表达的影响[J].中国现代医学杂志,2015,25(34):6-9.