朱昌毫;孙诚谊;喻超;田舍;张乙凡;陈世裕;潘耀振;
ZHU Changhao;SUN Chengyi;YU Chao;TIAN She;ZHANG Yifan;CHEN Shiyu;PAN Yaozhen;Department of Hepatobiliary Surgery,the Affiliated Hospital of Guizhou Medical University;Key Laboratory of Hepatobiliary and Pancreatic Surgery,Guizhou Medical University;

• 1:贵州医科大学附院肝胆外科
• 2:贵州医科大学肝胆胰脾重点实验室

摘要(Abstract)：

目的:体外实验探讨青藤碱对胰腺癌细胞PANC-1侵袭和迁移能力的影响。方法:用高糖培养基DMEM培养胰腺癌PANC-1细胞,取对数期生长的细胞,实验组用0.5、1.0 mmol/L青藤碱处理,运用侵袭和迁移实验研究青藤碱对胰腺癌侵袭转移的影响,采用激光共聚焦实验观察青藤碱对细胞骨架的改变,免疫印迹技术检测青藤碱对胰腺癌细胞PANC-1 E-钙粘着蛋白Nimentin和波形蛋白(E-cadherin)表达水平的改变。结果:实验组PANC-1细胞穿过铺有Matrigel胶小室的细胞数量显著减少,差异有统计学意义(P<0.05);细胞划痕实验表明,青藤碱处理24 h和48 h的胰腺癌细胞迁移速度明显小于对照组(P<0.05);激光共聚焦显示,实验组PANC-1细胞内绿色荧光染色的F-actin与对照组比较显著减少,且荧光强度也较弱(P<0.05);β-微管蛋白(β-tubulin)红色荧光强度明显减弱,差异有统计学意义(P<0.05);免疫印迹结果显示,实验组Vimentin逐渐减少,E-cadherin逐渐增多,与对照组比较差异具有统计学意义(P<0.05)。结论:一定浓度的青藤碱可抑制体外胰腺癌PANC-1细胞的侵袭和转移能力。
Objective: To study the effect of sinomenine on invasion and migration of pancreatic cancer cell PANC-1. Methods: PANC-1 cells were cultured in vitro,and treated with different concentrations of sinomenine(0,0. 5,1. 0 mmol/L) at the growing period of cells,Transwell method and wound healing experiment were adopted to test the invasion and migration of Panc-1 cells; The influence of SIN on cytoskeleton was detected with immunofluorescence technique; The changes of EMT-associated protein levels of Panc-1 cells after exposed with different concentrations of SIN were detected by Western Blotting. Result: Transwell experiment indicated the passing Panc-1 cell numbers in experiment groups were obviously less than that in control group( P < 0. 05). Wound healing experiment discovered that in 24 h and 48 h after infection,Panc-1 cells in control group migrated obviously faster than those in experiment groups. Immunofluorescence test showed that in SIN untreated Panc-1 cells,the parallel arrangement and cross overlapping reticulated bundle fibers formed by F-actin microfilament appeared in the whole cell and a lot of microfilaments penetrated deeply in cell processes. In comparison with control group,the F-actin of experiment group which stained with FITC-phalloidin was obviously decreased,and the microfilaments in kytoplasm became shorter or disappeared or became rarefied and arranged irregularly. Compared with the control group,the length of the microtubules were much shorter,bundle structure disapperaed or ruptured and the red fluorescence intensity was significantly reduced. Westernblot indicated that the expression of vimentin was significantly decreased and E-cadherin greatly enhanced compared with the control group. Conclusion: These results suggest that sinomenine could inhibit the invision and migration of human pancreatic cancer cell line PANC-1.

关键词(KeyWords)： 胰腺肿瘤;植物药;青藤碱;侵袭;转移
pancreatic cancer;botanicals;sinomenine;invasion;migration

Abstract:

Keywords:

基金项目(Foundation): 国家国际科技合作专项资助项目(2014DFA31420);; 国家自然科学基金资助项目(81660483);; 贵州省科学技术基金资助项目[黔科合J字(2015)2013号];; 贵州省第四批人才基地基金资助项目[黔省专合字(2012)94号];; 贵州省科学技术厅-贵阳医学院院士工作站肝胆外科分站[黔科合院士站(2015)4013];; 贵州省高层次创新人才培养计划“十”层次人才[黔科合平台人才(2016)5647];; 贵州省孙诚谊“肝胆胰脾疾病诊治”研究生导师工作室[黔教研合GZS(2016)09];; 贵州省肝胆外科临床医学研究中心[黔科合平台人才(2017)5404]

作者(Author): 朱昌毫;孙诚谊;喻超;田舍;张乙凡;陈世裕;潘耀振;
ZHU Changhao;SUN Chengyi;YU Chao;TIAN She;ZHANG Yifan;CHEN Shiyu;PAN Yaozhen;Department of Hepatobiliary Surgery,the Affiliated Hospital of Guizhou Medical University;Key Laboratory of Hepatobiliary and Pancreatic Surgery,Guizhou Medical University;

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