CAJ | 학술논문

目的建立对吉西他滨耐药的胰腺癌PANC-1/Gem细胞株,检测诱导耐药前后该细胞株生物学特性的变化.探讨吉西他滨诱导胰腺癌耐约的可能机制.方法通过逐渐增加培养基中吉西他滨的浓度,建屯对吉西他滨耐药的胰腺癌PANC-1/Gem细胞株,TUNEL染色检测细胞株凋亡,MTT方法检测胰腺癌PANC-1和胰腺癌PANC-1/Gem细胞的半数抑制浓度(IC_(50))和耐药指数(RI),Western印迹法检测酸性神经磷脂酶表达变化、二酰基甘油激酶(diacylglycerol kinase,DAGK)法检测神经酰胺含量变化.结果经过24周成功诱导出对吉西他滨耐药的胰腺癌PANC-1/Gem细胞株,吉西他滨对PANC-1和PANC-1/Gem细胞的IC_(50)值分别为:亲本(8.13±0.85)μg/ml,24周(285.40±34.83)μg/ml,与亲本细胞相比耐药倍数为35.10倍,且凋亡率降低.Western印迹检测发现吉西他滨诱导24周的胰腺癌细胞PANC-1/Gem酸性神经磷脂酶的表达低于亲本细胞.两组细胞的神经酰胺含量分别为:亲本(364.95±46.11)pmol/mg protein,24周(120.61±20.07)pmol/mgprotein.结论酸性神经磷脂酶的表达降低,导致神经酰胺含量减少可能是胰腺癌PANC-1细胞株对吉西他滨产生耐药的机制之一.
목적 건립대길서타빈내약적이선암PANC-1/Gem세포주,검측유도내약전후해세포주생물학특성적변화.탐토길서타빈유도이선암내약적가능궤제.방법 통과축점증가배양기중길서타빈적농도,건둔대길서타빈내약적이선암PANC-1/Gem세포주,TUNEL염색검측세포주조망,MTT방법 검측이선암PANC-1화이선암PANC-1/Gem세포적반수억제농도(IC_(50))화내약지수(RI),Western인적법검측산성신경린지매표체변화、이선기감유격매(diacylglycerol kinase,DAGK)법검측신경선알함량변화.결과 경과24주성공유도출대길서타빈내약적이선암PANC-1/Gem세포주,길서타빈대PANC-1화PANC-1/Gem세포적IC_(50)치분별위:친본(8.13±0.85)μg/ml,24주(285.40±34.83)μg/ml,여친본세포상비내약배수위35.10배,차조망솔강저.Western인적검측발현길서타빈유도24주적이선암세포PANC-1/Gem산성신경린지매적표체저우친본세포.량조세포적신경선알함량분별위:친본(364.95±46.11)pmol/mg protein,24주(120.61±20.07)pmol/mgprotein.결론 산성신경린지매적표체강저,도치신경선알함량감소가능시이선암PANC-1세포주대길서타빈산생내약적궤제지일.
Objective To establish gemcitabine-resistant pancreatic cancer cell line PANC-1/ gem and discuss its biological characters and drug resistance mechanism. Methods gemcitabine-resist-ant pancreatic cancer cell line was obtained by culture of pancreatic cancer cell line PANC-1 in vitro with intermittently increasing the concentration of gemcitabine in the culture medium for 24 weeks. After that, TUNEL technique was employed to detect the apoptosis. Meanwhile, its drug sensitivity as well as the expression of sphingomyelinase, contents of C2-ceramide were determined respectively. Results Drug-resistant PANC-1/Gem cell was successfully induced with gemeitabine for 24 weeks. The IC_(50) increased from (8.13±0.85)μg/ml in PANC-1 to (285.40±34.83)μg/ml in PANC-1/Gem. Compared with the normal pancreatic cancer cell line PANC-1, the drug resistance indexes of PANC-1/gem to gemcitabine was 35.1 and the apoptosis rate was reduced from 6.21 to 2.74%. The ex-pression of sphingomyelinase was lower than that of pancreatic cancer cell line PANC-1, and the con-tents of ceramide was reduced from (364.95±46.11 pmol/mg protein) to (120.61±20.07 pmol/mg protein). Conclusion The durg resistance of gemcitabine to pancreatic cancer cell line is positively rel-evant to the low expression of acid sphingomyelinase as well as ceramide deficiency.