CAJ | 학술논문

目的：以人胰腺癌细胞系PANC-1为研究对象，利用不同浓度舒林酸处理PANC-1细胞，观察其对PANC-1细胞增殖、凋亡影响，并探讨舒林酸通过抑制Wnt/β-catenin信号通路杀伤PANC-1细胞的可能机制。方法实验分为阴性对照组（加入不含舒林酸的DMSO）和实验组（分别加入舒林酸浓度为0.25、0.5、1、1.5、2 mM的培养液，分别为：阴性对照组、0.25 mM组、0.5 mM组、1.0 mM组、1.5 mM组、2.0 mM组，总计6组）。MTT法检测PANC-1细胞生长抑制率；流式细胞术检测细胞凋亡率；RT-PCR和免疫细胞化学技术检测细胞内β-Catenin的表达。结果 MTT结果显示：PANC-1细胞经舒林酸干预后生长均受到不同程度抑制，且随着舒林酸浓度增加，抑制作用越强。流式细胞术检测凋亡结果显示PANC-1细胞早期凋亡率干预后均有不同程度增加，与对照组相比，0.5、1.0 mM组早期凋亡率差异无统计学意义，而1.5、2.0 mM组差异均有统计学意义（P＜0.05）。RT-PCR结果显示可见各组β-catenin mRNA表达量随着舒林酸浓度增加而降低，0.5、1.0 mM组与对照组相比差异无统计学意义，而1.5及2.0 mM组差异有统计学意义（P＜0.05）；应用2.0 mM的舒林酸处理PANC-1细胞0、12、24、48、72 h，随着处理时间延长，β-catenin mRNA表达量逐渐降低，各处理组与对照组相比，差异有统计学意义（P＜0.05）。ICC结果证实干预后PANC-1细胞内β-catenin表达量及核聚集降低，与对照组相比，0.25、0.5 mM组差异无统计学意义，而1.0、1.5、2.0 mM组差异有统计学意义（P＜0.05）。结论舒林酸对PANC-1细胞具有生长抑制及促凋亡作用，这种作用具有浓度-时间依赖性，舒林酸抑制Wnt/β-catenin信号通路可能是其杀伤PANC-1细胞的可能机制之一。
목적：이인이선암세포계PANC-1위연구대상，이용불동농도서림산처리PANC-1세포，관찰기대PANC-1세포증식、조망영향，병탐토서림산통과억제Wnt/β-catenin신호통로살상PANC-1세포적가능궤제。방법실험분위음성대조조（가입불함서림산적DMSO）화실험조（분별가입서림산농도위0.25、0.5、1、1.5、2 mM적배양액，분별위：음성대조조、0.25 mM조、0.5 mM조、1.0 mM조、1.5 mM조、2.0 mM조，총계6조）。MTT법검측PANC-1세포생장억제솔；류식세포술검측세포조망솔；RT-PCR화면역세포화학기술검측세포내β-Catenin적표체。결과 MTT결과현시：PANC-1세포경서림산간예후생장균수도불동정도억제，차수착서림산농도증가，억제작용월강。류식세포술검측조망결과현시PANC-1세포조기조망솔간예후균유불동정도증가，여대조조상비，0.5、1.0 mM조조기조망솔차이무통계학의의，이1.5、2.0 mM조차이균유통계학의의（P＜0.05）。RT-PCR결과현시가견각조β-catenin mRNA표체량수착서림산농도증가이강저，0.5、1.0 mM조여대조조상비차이무통계학의의，이1.5급2.0 mM조차이유통계학의의（P＜0.05）；응용2.0 mM적서림산처리PANC-1세포0、12、24、48、72 h，수착처리시간연장，β-catenin mRNA표체량축점강저，각처리조여대조조상비，차이유통계학의의（P＜0.05）。ICC결과증실간예후PANC-1세포내β-catenin표체량급핵취집강저，여대조조상비，0.25、0.5 mM조차이무통계학의의，이1.0、1.5、2.0 mM조차이유통계학의의（P＜0.05）。결론서림산대PANC-1세포구유생장억제급촉조망작용，저충작용구유농도-시간의뢰성，서림산억제Wnt/β-catenin신호통로가능시기살상PANC-1세포적가능궤제지일。
Objective To discuss the influence of different concentration sulindac on pancreatic cancer cell line PANC-1 cell proliferation and apoptosis,and investigate the possible mechanism that sulindac can inhibit the Wnt/beta-catenin pathway to kill pancreatic cancer cells. Methods PANC-1 cell were divided into negative control group (added containing no sulindac DMSO)and experimental group (added sulindac with concentrations of 0.25 ,0.5 ,1 ,1.5 ,2 mM medium,respectively,name as 0.25 mM group,0.5 mM group,1.0 mM group,1.5 mM group,2.0 mM group),and treated with different time,cell proliferation inhibition ratio in each group was detected by MTT assay,cell apoptosis ratio was detected by flow cytometry,the expression ofβ-catenin mRNA and protein were detected by RT-PCR and immunocytochemistry.Results MTT results showed that sulindac can inhibit the cell proliferation of PANC-1 by a dose-and time-dependent manner.Cell apoptosis increased after sulindac treatment in different degrees,and there were statistical differences between 1.5,2.0 mMgroup and control groups (P<0.05).RT-PCR results showed that the expression ofβ-catenin mRNA decreased after the treatment of sulindac,there were statistical differences between 1.5,2.0 mMgroup and control group (P<0.05). In the 2.0mM group,the expression ofβ-catenin decreased along with the time extending (P<0.05 ).ICC results showed that sulindac inhibited the expression ofβ-catenin protein and nuclear accumulation,there were no statistical differences in 0.25 ,0.5 mM group and control group,but there were statistical differences in 1.0,1.5,2.0 mMgroup.Conclusion Sulindac could inhibit cell proliferation and facilitate apoptosis of PANC-1,this effect is dose-and time-dependent.The inhibition of Wnt/beta-catenin signal pathway may be a possible mechanism of its cytotoxicity.