胰腺癌吉西他滨耐药细胞株的建立及其生物学行为分析  

Establishment and investigation of the biological behavior of gemcitabine-resistant pancreatic cancer cell line

作　　者：朱皓阳 田嘉玮 屈申奥 陶诗然 安怡荣 路璐[1] 刘畅[1] 吕毅[2] 张娜娜 Zhu Haoyang;Tian Jiawei;Qu Shenao;Tao Shiran;An Yirong;Lu Lu;Liu Chang;Lyu Yi;Zhang Nana(Department of Anesthesiology,the First Affiliated Hospital of Xi'an Jiaotong Unversity,Xi'an 710061,China;Department of Hepatobiliary Surgery,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China;National Local Joint Engineering Research Center for Precision Surgery&Regenerative Medicine,the First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China)

机构地区：[1]西安交通大学第一附属医院麻醉手术部,西安710061 [2]西安交通大学第一附属医院肝胆外科,西安710061 [3]西安交通大学第一附属医院精准外科与再生医学国家地方联合工程研究中心,西安710061

出　　处：《中华肝胆外科杂志》2025年第1期59-65,共7页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金(82202297);陕西省自然科学基础研究计划(2023-JC-QN-0905);西安交通大学第一附属医院科研发展基金(2021QN-23)。

摘　　要：目的构建人源胰腺癌细胞株PANC1与鼠源胰腺癌细胞株PANC02的吉西他滨耐药细胞株,并探究其生物学行为改变情况。方法采用浓度梯度递增法建立人源胰腺癌细胞PANC1及鼠源胰腺癌细胞PANC02的吉西他滨耐药细胞株,即PANC1-GR、PANC02-GR。采用细胞计数试剂(CCK-8)、流式细胞术、细胞划痕及Transwell实验检测四组细胞的耐药、增殖、细胞周期、迁移与侵袭等,并与亲本细胞比较。结果PANC1-GR及PANC02-GR的耐药指数分别为153.3和185.4。CCK-8检测结果显示耐药细胞的增殖速度明显增快,PANC1-GR较PANC1的细胞群体倍增时间缩短(1.5±0.1)d比(2.4±0.2)d,差异有统计学意义(t=8.00,P<0.001)。流式细胞仪结果显示两种耐药细胞处于S期与G2/M期的细胞比例增加,G0/G1期的比例降低。细胞划痕及Transwell实验显示,与亲本细胞相比,PANC1-GR和PANC02-GR细胞的24 h迁移率增高(47.6±2.4)%比(28.7±6.3)%、(53.6±3.2)%比(30.1±1.4)%,穿膜数量(放大200倍的单个视野)也增加(269.7±30.9)个比(62.7±10.1)个、(172.0±30.8)个比(36.3±4.9)个,差异均有统计学意义(均P<0.05)。结论浓度梯度递增法可建立胰腺癌吉西他滨耐药细胞株,耐药细胞株的增殖能力、迁移、侵袭性比亲本更强,为耐药机制研究提供基础。ObjectiveTo construct the gemcitabine resistant cell lines of human pancreatic cancer cell line(PANC1)and mouse pancreatic cancer cell line(PANC02),and to investigate their biological behavior changes.MethodsGemcitabine-resistant cell lines PANC1-GR of human pancreatic cancer and PANC02-GR of mouse pancreatic cancer were induced by concentration gradient increment method.Cell count assay(CCK-8),flow cytometry,cell scratch assay and Transwell assay were used to detect the drug resistance,proliferation,cell cycle,migration and invasion of the four groups of cell lines.The drug-resistant cells were also compared with the parent cells.ResultsThe resistance indices of PANC1-GR and PANC02-GR were 153.3 and 185.4,respectively.The results of CCK-8 showed that with the increase of gemcitabine concentration,the proliferation of resistant cells changed significantly compared with parental cells,the population doubling time of PANC1-GR was significantly shorter than that of PANC1(1.5±0.1)d vs(2.4±0.2)d(t=8.00,P<0.001).The proportion of cells in S and G2/M phase increased,and the proportion of cells in G0/G1 phase decreased.The cell scratch and Transwell experiments indicated that the 24h mobility of PANC1-GR and PANC02-GR was higher than that of parent cells(47.6±2.4)%vs(28.7±6.3)%and(53.6±3.2)%vs(30.1±1.4)%,the number of individual field(200 times magnification)penetrating membrane cells was also higher than that of parent cells(269.7±30.9)vs(62.7±10.1)and(172.0±30.8)vs(36.3±4.9),with statistical significance(all P<0.05).ConclusionConcentration gradient increment method can successfully establish gemcitabine-resistant pancreatic cancer cell lines,which have stronger proliferation,migration and invasiveness,and can be used to study the mechanism of drug resistance in pancreatic cancer.

关 键 词：胰腺肿瘤 抗药性 肿瘤 吉西他滨 生物学行为 

分 类 号：R735.9[医药卫生—肿瘤]