学术论文

      AG490对 Janus 蛋白酪氨酸激酶与信号转导和转录激活子3调控的胆囊癌 SGC-996细胞株侵袭和转移的影响

      AG490 regulated the invasion and metastasis of gallbladder SGC -996 cells through JAK/STAT3 pathway

      摘要:
      目的:探讨 AG490对 Janus 蛋白酪氨酸激酶与信号转导和转录激活子3(JAK/STAT3)调控的胆囊癌 SGC-996细胞株侵袭和转移的影响,并探讨其相关作用机制。方法采用 MTT 法检测50μmol/L、100μmol/L 和200μmol/L 不同浓度 AG490对 SGC-996细胞活力的影响,Transwell 评价 AG490对胆囊癌侵袭转移能力,流式细胞术检测 SGC-996细胞凋亡,Western blot 检测 SGC-996细胞 JAK/STAT3通路相关蛋白表达。结果不同浓度的 AG490对 SGC-996细胞活力抑制率分别为(17.49±3.41)%、(38.66±4.57)%、(79.15±6.29)%,且随着浓度的增加细胞活力明显下降,与对照组的(1.39±0.21)%差异均有统计学意义(t =8.162、14.111、21.401,均 P <0.01)。不同浓度的 AG490降低 SGC-996细胞转移抑制率[分别为(23.18±4.53)%、(51.75±6.46)%、(81.32±7.13)%],且随着 AG490浓度的增加抑制侵袭转移能力增强,与对照组的(1.46±0.42)%差异均有统计学意义(t =8.269、13.455、19.366,均 P <0.01)。不同浓度的AG490诱导 SGC-996细胞的凋亡促进率分别为(13.34±4.33)%、(28.16±6.23)%、(53.61±8.74)%,且随着浓度的增加细胞凋亡增高,与对照组的(0.97±0.52)%差异均有统计学意义(t =4.913、7.533、10.414,均P <0.01)。不同浓度的 AG490降低 SGC-996细胞 JAK/STAT3通路的 ZFX、STAT3和 Smad1蛋白表达,与对照组差异均有统计学意义(tZFX =2.154、3.041、4.185,tSTAT3=7.348、14.892、17.774,tSmad1=3.474、5.241、7.718,均 P <0.05)。结论 AG490可抑制胆囊癌 SGC-996细胞的侵袭和转移,且呈剂量依赖性,其作用机制可能与降低细胞凋亡和 JAK/STAT3通路蛋白表达有关。
      Abstract:
      Objective To study the effect of AG490 on the invasion and metastasis of gallbladder SGC -996 cells regulated by JAK/STAT3,and discuss the related mechanism.Methods Cell viability treated with different concentrations of AG490(50,100,200μmol/L)was detected by MTT method.The ability of invasion and metastasis of gallbladder cells was evaluated by Transwell membrane count.The SGC -996 cell apoptosis was detected by flow cytometry.The protein expression of JAK/STAT3 pathway was detected by Western blot.Results The viability inhi-bition rates of different concentrations of AG490 for SGC -996 cells were (17.49 ±3.41)%,(38.66 ±4.57)%, (79.15 ±6.29)% respectively,and with the increasing of concentration,cell viability decreased obviously.Compared with the control group[(1.39 ±0.21)%],the differences were statistically significant(t =8.162,14.111,21.401, all P <0.01 ).The transfer ability inhibition rate of different concentrations of AG490 for SGC -996 cell were (23.18 ±4.53)%,(51.75 ±6.46)%,(81.32 ±7.13)% respectively,and with the increasing of concentration of AG490,the inhibition rate of invasion and metastasis enhanced.Compared with the control group [(1.46 ± 0.42)%],the differences were statistically significant(t =8.269,13.455,19.366,all P <0.01).The apoptosis rate for SGC -996 cells of different concentrations of AG490 were (13.34 ±4.33)%,(28.16 ±6.23)%,(53.61 ±&nbsp;8.74)% respectively,and cell apoptosis increased with the increasing of concentration.Compared with control group [(0.97 ±0.52)%],the differences were statistically significant(t =4.913,7.533,10.414,all P <0.01).Different concentrations of AG490 can reduce expression of ZFX,STAT3 and Smad1 protein of JAK/STAT3 pathway of SGC -996 cells,compared with the control group,the differences were statistically significant(tZFX =2.154,3.041,4.185, tSTAT3 =7.348,14.892,17.774 and tSmad1 =3.474,5.241,7.718,all P <0.05).Conclusion AG490 can inhibit the invasion and metastasis of gallbladder SGC -996 cells,and the effect depends on dosage.Its mechanism may be relat-ed to the reduction of cell apoptosis and the protein expression of JAK/STAT3 pathway.
      Author: Shi Hua Wang Jianbo Pan Jundi Ye Bin
      作者单位: 丽水市中心医院消化科, 浙江省丽水,323000
      刊 名: 中国基层医药 ISTIC
      年,卷(期): 2017, 24(6)
      在线出版日期: 2017年3月29日
      基金项目: 浙江省科技计划项目(2015C33288)Fund programScience and technology planning project of Zhejiang Province