摘要
目的探讨布托啡诺(Btpn)调节缺氧诱导因子-1α(HIF-1α)/血管内皮生长因子(VEGF)信号通路对子宫内膜癌细胞迁移、侵袭和血管生成的影响。方法以人子宫内膜癌Ishikawa细胞为研究对象,用不同浓度Btpn(0、2.5、5、10、20、40、80μg/ml)处理Ishikawa细胞,CCK-8法检测Ishikawa细胞增殖,并计算半抑制浓度(IC50)。取对数生长期的Ishikawa细胞,将其分为对照组(NC组)、低剂量Btpn组(Btpn-L组,5μg/ml)、中剂量Btpn组(Btpn-M组,10μg/ml)、高剂量Btpn组(Btpn-H组,20μg/ml)、DMOG(HIF-1α激活剂)组(10μmol/L)、Btpn-H+DMOG组(20μg/ml+10μmol/L)。划痕实验检测Ishikawa细胞迁移能力;Transwell实验检测Ishikawa细胞侵袭能力;Matrigel基质胶法检测Ishikawa细胞管腔形成;Western blot检测Ishikawa细胞中基质金属蛋白酶(MMP)-2、MMP-9、环氧化酶-2(COX-2)、HIF-1α、VEGF蛋白表达。结果与0μg/ml Btpn比较,2.5、5、10、20、40、80μg/ml Btpn处理的Ishikawa细胞OD450值降低(P<0.05),经计算得IC50值为19.24μg/ml,因此,选取5、10、20μg/ml Btpn作为后续处理Ishikawa细胞的低、中、高剂量浓度。与NC组比较,Btpn-L组、Btpn-M组、Btpn-H组Ishikawa细胞划痕愈合率、细胞侵袭数目、管腔形成数、MMP-2、MMP-9、COX-2、HIF-1α、VEGF蛋白表达降低,DMOG组Ishikawa细胞划痕愈合率、细胞侵袭数目、管腔形成数、MMP-2、MMP-9、COX-2、HIF-1α、VEGF蛋白表达升高(P<0.05);与Btpn-H组比较,Btpn-H+DMOG组Ishikawa细胞划痕愈合率、细胞侵袭数目、管腔形成数、MMP-2、MMP-9、COX-2、HIF-1α、VEGF蛋白表达升高(P<0.05)。结论Btpn可能通过抑制HIF-1α/VEGF信号通路抑制Ishikawa细胞迁移、侵袭和血管生成。
Objective To investigate the regulatory effects of butorphanol(Btpn)on migration,invasion and angiogenesis of endometrial carcinoma cells by regulating the hypoxia inducible factor-1α(HIF-1α)/vascular endothelial growth factor(VEGF)signaling pathway.Methods The human endometrial carcinoma cell line Ishikawa was induced with 0,2.5,5,10,20,40 and 80μg/ml Btpn,followed by detecting cell proliferation and calculating the half maximal inhibitory concentration(IC50)via cell counting kit-8(CCK-8)assays.Ishikawa cells in logarithmic growth phase were treated with blank control(NC group),low-dose Btpn at 5μg/ml(Btpn-L group),medium-dose Btpn at 10μg/ml(Btpn-M group),high-dose Btpn at 20μg/ml(Btpn-H group),10μmol/L HIF-1αactivator DMOG(DMOG group),and 20μg/ml Btpn+10μmol/L DMOG(Btpn-H+DMOG group).Cell migration,invasion and lumen formation were detected by wound healing assay,Transwell assay and ECM gel matrix assay,respectively.Protein expressions of matrix metalloproteinase-2(MMP-2),MMP-9,cyclooxygenase-2(COX-2),HIF-1αand VEGF in Ishikawa cells were detected by Western blot.Results Compared with 0μg/ml Btpn,OD450 value in Ishikawa cells induced with 2.5,5,10,20,40,80μg/ml Btpn significantly decreased(P<0.05),and the IC50 value was calculated as 19.24μg/ml.Therefore,5,10,and 20μg/ml were selected as the low,medium and high doses of Btpn for the following experiments,respectively.Compared with NC group,the wound healing rate,number of invasive cells,number of formed lumens,and protein expressions of MMP-2,MMP-9,COX-2,HIF-1αand VEGF in Ishikawa cells of Btpn-L,Btpn-M and Btpn-H group were significantly reduced(P<0.05),which in DMOG group were significantly higher(P<0.05).Compared with Btpn-H group,the wound healing rate,number of invasive cells,number of formed lumens,and protein expressions of MMP-2,MMP-9,COX-2,HIF-1αand VEGF in Ishikawa cells of Btpn-H+DMOG group were significantly higher(P<0.05).Conclusion Btpn may inhibit migration,invasion and angiogenesis of Ishikawa cells by inhibiting the HIF-1α/VE
作者
刘庆冬
宋美微
袁魁
LIU Qingdong;SONG Meiwei;YUAN Kui(Department of Anesthesiology,Tumour Hospital of Mudanjiang City,Heilongjiang,Mudanjiang 157009,China;不详)
出处
《河北医药》
CAS
2023年第14期2095-2099,共5页
Hebei Medical Journal
基金
黑龙江省中医药科研项目(编号:ZHY19-040)。
关键词
子宫内膜癌
布托啡诺
缺氧诱导因子-1α/血管内皮生长因子信号通路
迁移
侵袭
血管生成
endometrial carcinoma
butorphanol
hypoxia inducible factor-1α/vascular endothelial growth factor signaling pathway
migration
invasion
angiogenesis
