金圣草黄素抑制IL-6/JAK2/STAT3通路抑制人胃癌裸鼠原位移植瘤生长和转移

Chrysoeriol Inhibits the Growth and Metastasis of Orthotopic Grafted Human Gastric Cancer in Nude Mice by Inhibiting IL-6/JAK2/STAT3 Pathway

目的:探讨金圣草黄素(Chrysoeriol,Chry)对人胃癌裸鼠原位移植瘤生长和转移及IL-6/JAK2/STAT3信号通路的影响。方法:建立人胃癌裸鼠原位移植模型,将造模成功裸鼠随机分为模型组(Model组)、金圣草黄素低、中、高剂量组(Chry-L、Chry-M、Chry-H组)、金圣草黄素高剂量+JAK2激活剂Coumermycin A1组(Chry-H+CA1组),每组12只。皮下注射给药结束后处死裸鼠取移植瘤并称重,计算抑瘤率;HE染色观察移植瘤组织病理变化情况;TUNEL染色检测移植瘤组织细胞凋亡情况;免疫组化检测移植瘤迁移与侵袭相关蛋白基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)蛋白表达情况;免疫印迹检测移植瘤细胞凋亡及IL-6/JAK2/STAT3信号通路相关蛋白表达情况。结果:与Model组相比,Chry-L、Chry-M、Chry-H组肿瘤细胞密度降低,排列疏松紊乱,大量肿瘤细胞破裂坏死,结缔组织增生明显,核固缩深染增多,核分裂相减少,凋亡坏死细胞增多,移植瘤质量减轻,抑瘤率、细胞凋亡指数及Bax表达升高,肿瘤转移率及Bcl-2、MMP-2、MMP-9、IL-6、p-JAK2/JAK2、p-STAT3/STAT3表达降低,Chry-H组变化最明显(P<0.05);与Chry-H组相比,Chry-H+CA1组肿瘤细胞增多,排列相对整齐,核固缩深染减少,核分裂相增多,肿瘤坏死灶明显,移植瘤质量增重,抑瘤率、细胞凋亡指数及Bax表达降低,肿瘤转移率及Bcl-2、MMP-2、MMP-9、IL-6、p-JAK2/JAK2、p-STAT3/STAT3表达升高(P<0.05)。结论:金圣草黄素可抑制人胃癌裸鼠原位移植瘤生长和转移,其作用机制与抑制IL-6/JAK2/STAT3信号通路有关。

Objective:To investigate the effects of Chrysoeriol(Chry)on the growth and metastasis of human gastric cancer in nude mice and the IL-6/JAK2/STAT3 signaling pathway.Methods:Orthotopic transplantation model of human gastric cancer nude mice was established,and successfully modeled nude mice were randomly separated into a model group,low,medium,and high dose Chry groups(Chry-L,Chry-M,Chry-H groups),and high-dose Chry+JAK2 activator Coumermycin A1 group(Chry-H+CA1 group),12 animals per group.After subcutaneous injection,the nude mice were killed to take transplanted tumors and weigh them,and the tumor inhibition rate was calculated.HE staining was applied to observe the pathological changes of transplanted tumor tissue.TUNEL staining was applied to detect apoptosis in transplanted tumor tissues.Immunohistochemistry was applied to detect the expression of matrix metalloproteinase-2(MMP-2)and matrix metalloproteinase-9(MMP-9)proteins related to migration and invasion of transplanted tumors.Immunoblotting was applied to detect apoptosis of transplanted tumor cells and the expression of IL-6/JAK2/STAT3 signaling pathway related proteins.Results:Compared with the model group,the tumor cell density in the Chry-L,Chry-M,and Chry-H groups decreased,with loose and disordered arrangement,a large number of tumor cells ruptured and necrotic,obvious proliferation of connective tissue,with an increase in nuclear pyknosis and deep staining,a decrease in nuclear division,an increase in apoptotic and necrotic cells,and a reduction in the quality of transplanted tumors,the tumor inhibition rate,cell apoptosis index,and Bax expression increased,the tumor metastasis rate and the expression of Bcl-2,MMP-2,MMP-9,IL-6,p-JAK2/JAK2,p-STAT3/STAT3 reduced,the Chry-H group showed the most obvious changes(P<0.05).Compared with the Chry-H group,the Chry-H+CA1 group showed an increase in tumor cells,relatively neat arrangement,reduced nuclear pyknosis and deep staining,increased nuclear division,obvious tumor necrosis,and increased weight of transplanted tumors,the tumor inhibition rate,cell apoptosis index,and Bax expression decreased,the tumor metastasis rate and the expression of Bcl-2,MMP-2,MMP-9,IL-6,p-JAK2/JAK2,and p-STAT3/STAT3 increased(P<0.05).Conclusion:Chrysoeriol can inhibit the growth and metastasis of human gastric cancer transplanted in nude mice,and its mechanism of action is related to the inhibition of the IL-6/JAK2/STAT3 signaling pathway.