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PANCl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、287nmol／L人参炔醇处理干细胞12、24、48h，采用CCK8法检测细胞成活率；通过克隆形成实验观察287nmol／L人参炔醇处理胰腺癌干细胞48h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B—catenin的表达。结果经干细胞培养体系培养的胰腺癌PANCl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48h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287nmol／L人参炔醇处理48h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、B—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PANCl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。
Objective 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. Cuhured 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 13-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 stermellsincontro1,72,144,287nmol/Lpanaxynolgroupwas100%, （ 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 13-eatenin 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 downregnlating Ki67, PCNA and blocking Wnt/β-catenin signaling pathway,
出处
《中华胰腺病杂志》
CAS
2016年第4期221-224,共4页
Chinese Journal of Pancreatology
基金
国家自然基金（81502663）
江苏省社会发展项目（BE2015668）
江苏省高校自然基金（14KJD310001）
江苏大学临床重点专项基金（JDLCZX005）
镇江市社会发展项目（SH2014053）
关键词
胰腺肿瘤
干细胞
人参炔醇
细胞增殖
Pancreatic neoplasms
Stem cells
Panaxynol
Cell proliferation
