摘要目的 探讨人参炔醇对胰腺癌干细胞增殖及自我更新的影响.方法 胰腺癌PANC1细胞在干细胞体系培养液中培养成干细胞,用流式细胞术检测胰腺癌干细胞的CD133+细胞比例.以72、144、287 nmol/L人参炔醇处理干细胞12、24、48 h,采用CCK8法检测细胞成活率;通过克隆形成实验观察287 nmol/L人参炔醇处理胰腺癌干细胞48 h后的集落数量;采用蛋白质印迹法检测细胞增殖相关蛋白Ki67、PCNA及自我更新相关蛋白β-catenin的表达.结果 经干细胞培养体系培养的胰腺癌PANC1细胞的CD133+细胞比例为(9.70±0.59)％,显著高于常规培养对照组的(2.11±0.25)％,差异有统计学意义(P＜0.001).人参炔醇呈浓度及时间依赖性减少胰腺癌干细胞的存活率,对照组及72、144、287nmol/L人参炔醇处理48 h组细胞的存活率分别为100％、(63.32± 2.37)％、(49.91±2.13)％、(41.37±2.01)％,差异有统计学意义(P＜0.001);对照组及287 nmol/L人参炔醇处理48 h组细胞的集落形成数分别为(611 ±25)、(280±16)个,人参炔醇处理组集落形成数显著减少,差异有统计学意义(P ＜0.001);对照组细胞Ki67、PCNA、β-catenin表达量分别为0.376±0.012、0.772±0.026、0.219±0.018,人参炔醇处理组分别为0.183±0.010、0.453 ±0.009、0.148 ±0.006,处理组的蛋白表达量均较对照组显著下降,差异有统计学意义(P值均＜0.001).结论 人参炔醇可抑制胰腺癌PANC1干细胞的增殖及自我更新,其机制可能与下调Ki67、PCNA表达,阻断Wnt/β-catenin信号通路有关.

abstractsObjective To investigate the influence of panaxynol on pancreatic cancer stem cells' proliferation and self-renewal.Methods PANC1 cells were cultured in stem cell culture system to induce the formation of stem cells,and the proportion of CD133 + pancreatic cancer stem cells was detected by FCM.Cultured pancreatic stem cells were treated with panaxynol at different concentrations of 0,72,144,287 nmol/L for 0,12,24,48 h.CCK8 kit was used to detect the cell survival.The colony formation experiment detected the number of colonies after being cultured with 287 nmol/L panaxynol for 48 h.Western blot was used to detect the expression of proliferation-related protein Ki67,PCNA and self-renewal related protein 3-catenin.Results The CD133 + proportion of pancreatic cancer stem cells was (9.70 ± 0.59) ％,which was statistically higher than that [(2.11 ±0.25)％] in the control group (P ＜0.001).Panaxynol can decrease the survival rates of pancreatic cancer stem cells in a dosage and time dependent manner.The survival rate of stemcellsincontrol,72,144,287nmol/Lpanaxynolgroupwas 100％,(63.32 ± 2.37) ％,(49.91 ± 2.13) ％ and (41.37 ± 2.01)％ after cultured for 48 h,which had statistically significant difference among different groups (P ＜ 0.001).The number of colonies in the control and 287 nmol/L panaxynol group was (611 ± 25) and (280 ± 16).Colonies in panaxynol group were fewer than those in the control group with statistically difference (P＜0.001).The expression of Ki67,PCNA and β-catenin were 0.376± 0.012,0.772± 0.026 and 0.219 ± 0.018 in the control group and were 0.183 ± 0.010,0.453 ± 0.009 and 0.148 ± 0.006 in panaxynol group,respectively.The results indicated that Ki67,PCNA and β-catenin were down-regulated by panaxynol treatment and the differences were statistically significant (P ＜ 0.01).Conclusions Panaxynol can inhibit the proliferation and self-renewal of pancreatic cancer stem cells.These effects may be related to downregulating Ki67,PCNA and blocking Wnt/β-catenin signaling pathway.