柚皮素对胰腺癌细胞PANC-1增殖和凋亡的影响  

Effects of naringin on the proliferation and apoptosis of the human pancreatic cancer cell line PANC-1

作　　者：夏莹 王世全 路伟 XIA Ying;WANG Shiquan;LU Wei(Department of Gastroenterology,First Hospital Affiliated to Air Force Medical University,Shaanxi,Xi’an 710032,China)

机构地区：[1]中国人民解放军空军军医大学第一附属医院消化内科,西安市710032

出　　处：《河北医药》2023年第13期1925-1929,共5页Hebei Medical Journal

基　　金：陕西省自然科学基金项目(编号:2020JM-330)。

摘　　要：目的探讨柚皮素对胰腺癌细胞PANC-1增殖和凋亡的影响及糖原合成酶激酶-3β(GSK-3β)在其中的作用。方法PANC-1细胞根据随机数字表分为7组(每组12孔,细胞密度为2500个/mm^(2)):0μmol/L、50μmol/L、100μmol/L、200μmol/L、400μmol/L、control virus和GSK-3βvirus组。0μmol/L组,加入相应体积溶剂;50μmol/L、100μmol/L、200μmol/L和400μmol/L分别在培养液中加入终浓度为50μmol/L、100μmol/L、200μmol/L和400μmol/L的柚皮素进行培养;Control virus组,感染了对照病毒的PANC-1细胞培养液中加入200μmol/L的柚皮素进行培养;GSK-3β组,感染了GSK-3β过表达慢病毒的PANC-1细胞培养液中加入200μmol/L柚皮素。CCK-8检测细胞活力,细胞计数仪测定细胞数量,流式细胞测量凋亡细胞百分比,Western blot方法检测Ki-67、cleaved caspase-3、GSK-3β和p-GSK-3β的蛋白表达水平;RT-PCR法测定GSK-3β的mRNA表达水平;免疫荧光法检测Ki-67阳性细胞数量。结果与0μmol/L组比较,100μmol/L、200μmol/L和400μmol/L组细胞活力、细胞增殖倍数、Ki-67阳性细胞比例、Ki67蛋白表达水平、GSk-3β和p-GSK-3β表达水平降低(P<0.05),细胞凋亡百分比和cleaved caspase-3表达量增加(P<0.05);与100μmol/L组比较,200μmol/L和400μmol/L组细胞活力、细胞增殖倍数、Ki-67阳性细胞比例、Ki67蛋白表达水平、GSk-3β和p-GSK-3β表达水平进一步下降(P<0.05),细胞凋亡百分比和Cleaved caspase-3表达量进一步增加(P<0.05);200μmol/L与400μmol/L组相比以上指标均无差异(P>0.05);与NA+control virus组相比较,NA+GSK-3β组Ki-67阳性细胞比例、Ki67蛋白表达水平增加(P<0.05),细胞凋亡百分比和Cleaved caspase-3表达量下降(P<0.05)。结论柚皮素通过降低GSK-3β的表达及活性抑制PANC-1细胞增殖并促进细胞凋亡。Objective To explore the effects of naringenin on the proliferation and apoptosis of the PANC-1 human pancreatic cancer cell line and the involvement of glycogen synthase kinase-3β(GSK-3β).Methods PANC-1 cells were seeded with 2.5×10^(3) cells/mm 2,and 12 replicates were prepared per group.Cells were incubated with vehicle,50μmol/L naringenin,100μmol/L naringenin,200μmol/L naringenin,400μmol/L naringenin,200μmol/L naringenin+control virus,and 200μmol/L naringenin+overexpression lentivirus of GSK-3β.Cell viability,cell number and apoptotic rate were detected by CCK-8 assay,cell counter and flow cytometry,respectively.Protein expressions of Ki-67,cleaved caspase-3,GSK-3βand p-GSK-3βwere detected by Western blot.The mRNA level of GSK-3βwas detected by RT-qPCR.The number of Ki-67-positive cells was detected by immunofluorescence.Results Compared with those in PANC-1 cells incubated with vehicle,cell viability,proliferation rate,proportion of Ki-67-positive cells,and protein expressions of Ki-67,GSK-3βand p-GSK-3βwere significantly reduced in those incubated with 100μmol/L,200μmol/L and 400μmol/L naringenin(P<0.05),while the apoptotic rate and the protein expression of cleaved caspase-3 were significantly enhanced(P<0.05).Compared with PANC-1 cells incubated with 100μmol/L naringenin,cell viability,proliferation rate,proportion of Ki-67-positive cells,and protein expressions of Ki-67,GSK-3βand p-GSK-3βwere significantly reduced in those incubated with 200μmol/L and 400μmol/L naringenin,while the apoptotic rate and the protein expression of cleaved caspase-3 were significantly enhanced(P<0.05).The above indexes were comparable between PANC-1 cells incubated with 200μmol/L and 400μmol/L naringenin(P>0.05).Compared with those incubated with 200μmol/L naringenin+control virus,the proportion of Ki-67-positive cells and protein expressions of Ki-67 were significantly enhanced in those incubated with 200μmol/L naringenin+overexpression lentivirus of GSK-3β,while the apoptotic rate and the protein

关 键 词：柚皮素 PANC-1 增殖 凋亡 

分 类 号：R735.9[医药卫生—肿瘤]