马蔺子甲素对人胰腺癌细胞株PANC-1增殖、迁移、侵袭的影响及其机制

Effects of irisquinone on proliferation,invasion,and migration of pancreatic ductal adenocarcinoma cell line PANC-1

目的观察马蔺子甲素(IQ)对人胰腺导管腺癌(PDAC)细胞株PANC-1增殖、侵袭、迁移的影响,并探讨其作用机制。方法取对数生长期的PANC-1细胞置于基础培养基中,将细胞分为高剂量组、中剂量组、低剂量组、对照组,高剂量组、中剂量组、低剂量组分别加入25、20、15μmol/L的IQ,对照组加入等量0.9%NaCl溶液。采用MTT比色法检测细胞增殖能力(细胞OD值),划痕愈合实验检测细胞迁移能力(细胞迁移距离),Transwell侵袭实验检测细胞侵袭能力(穿膜细胞个数),qPCR法检测细胞Rho信号通路关键分子RhoA、Rac1、ROCK1、CDC42、ROCK2。结果与对照组比较,中剂量组和高剂量组培养24、48、72 h的OD值减小(P均<0.05);与同组0 h比较,低剂量组及中剂量组和高剂量组培养24、48、72 h的OD值增加(P均<0.05)。与对照组比较,低剂量组、中剂量组、高剂量组细胞迁移距离短,穿膜细胞个数少(P均<0.05);与低剂量组比较,中剂量组、高剂量组细胞迁移距离短,穿膜细胞个数少(P均<0.05);与中剂量组比较,高剂量组细胞迁移距离短,穿膜细胞个数少(P均<0.05)。与对照组比较,低剂量组及中剂量组和高剂量组RhoA、Rac1、ROCK1相对表达量减小(P均<0.05);与低剂量组比较,中剂量组、高剂量组ROCK1相对表达量减小(P均<0.05)。结论IQ可抑制PANC-1细胞增殖、侵袭及转移,其作用机制可能为抑制Rho信号通路介导的上皮间质转化进程,当药物浓度为25μmol/L时可达到最大抑瘤效应。

Objective To observe the effects of irisquinone(IQ)on the proliferation,invasion,and migration of hu⁃man pancreatic ductal adenocarcinoma(PDAC)cell line PANC-1,and to investigate its mechanism.Methods PANC-1 cells in the logarithmic growth phase were cultured in basic culture medium and then were divided into high-dose group,medium-dose group,low-dose group and control group;25,20,and 15μmol/L IQ was added to the high-dose group,medium-dose group and low-dose group,respectively,while the control group was incubated with equal amount of 0.9%NaCl.The proliferation capacity of the cells was measured by MTT assay(cell OD value),the cell migration capaci⁃ty(cell migration distances)by Wound healing assay,and cell invasion capacity(the number of cells penetrating the mem⁃brane)by Transwell assay.The expression levels of RhoA,Rac1,ROCK1,CDC42 and ROCK2,which are key molecules in the Rho signaling pathway,were measured by qPCR.Results Compared with the control group,the OD values of the medium-dose and high-dose groups decreased at 24,48 and 72 h of incubation(all P<0.05).OD values increased in the low-dose group and the medium-dose and high-dose groups at 24,48 and 72 h of incubation in comparison with those of the same group at 0 h(all P<0.05).Compared with the control group,the low-dose,medium-dose and high-dose groups had shorter migration distances and fewer cells penetrating the membrane(all P<0.05).Compared with the lowdose group,the medium-dose and high-dose groups had shorter migration distances and fewer cells penetrating the mem⁃brane(all P<0.05).Compared with the medium-dose group,the high-dose group had a shorter migration distance and few⁃er cells penetrating the membrane(both P<0.05).Compared with the control group,the relative expression levels of RhoA,Rac1 and ROCK1 decreased in the low-dose,medium-dose and high-dose groups(all P<0.05);compared with the low-dose group,the relative expression of ROCK1 decreased in the medium-dose and high-dose groups(all P<0.05).Conclusion IQ can inhibit the proliferation,invasion,and migration of PANC-1 cells,and its mechanism of action may be the inhibition of the epithelial mesenchymal transition process mediated by Rho signaling pathway,and the maximum tu⁃mor suppressive effect can be achieved at a drug concentration of 25μmol/L.