Effect of gemcitabine on the expression of a poptosis-related genes in human pancreatic cancer cells
Effect of gemcitabine on the expression of a poptosis-related genes in human pancreatic cancer cells
复制标题
DOI：
10.3748/wjg.v12.i10.1597
复制
发表时间：
2006-03-14
影响因子：
4.3
通讯作者：
Minamoto, Toshinari
中科院分区：
文献类型：
--
作者：
Jiang, Pei-Hong;Motoo, Yoshiharu;Minamoto, Toshinari
目的：目的：研究吉西他滨对人胰腺癌细胞株PANC-1细胞毒作用相关基因的表达。将1 × 104个PANC-1细胞接种在96孔微量滴定板中。培养24 h后，加入浓度为2.5 ~ 1000 mg/L的吉西他滨。使用AlamarBlue染料法进行细胞生长分析。使用DNA片段化酶联免疫吸附测定（ELISA）试剂盒定量测定DNA片段化。采用逆转录-聚合酶链反应半定量分析PAP和TP 53 INP 1 mRNA的表达。结果：吉西他滨作用48 h后，对GSK-3 β和磷酸化GSK-3 β蛋白的IC 50为16 mg/L。吉西他滨对PANC-1细胞的生长呈浓度依赖性抑制（P < 0.0001），并在一定时间内抑制细胞生长（P < 0.0001）。吉西他滨治疗组48 h的DNA断裂率为44.7%，而未治疗组为25.3%。经吉西他滨处理后，PAP mRNA的表达降低，而TP 53 INP 1 mRNA的表达增加。结论：吉西他滨可抑制PANC-1细胞增殖并诱导其凋亡。细胞凋亡被认为与PAP和GSK-3 β的抑制以及TP 53 INP 1和pospho-GSK-3 β（ser 9）的激活有关。(C)2006年，WJG出版社。All rights reserved.
AIM: To investigate the expression of genes involved in the gemcitabine-induced cytotoxicity in human pancreatic cancer cells.METHODS: A human pancreatic cancer cell line, PANC-1, was cultured. 1 X 10(4) PANC-1 cells were plated in 96-well microtiter plates. After being incubated for 24 h, gemcitabine was added to the medium at concentrations ranging 2.5 -1 000 mg/L. The AlamarBlue dye method was used for cell growth analysis. DNA fragmentation was quantitatively assayed using a DNA fragmentation enzyme-linked immunosorbent assay (ELISA) kit. PAP and TP53INP1 mRNA expression was determined using the reverse transcription-polymerase chain reaction with semi-quantitative analysis. The expression of GSK-3 beta and phospho-GSK-3 beta proteins was examined with Western blot analysis.RESULTS: The IC50 for the drug after a 48-h exposure to gemcitabine was 16 mg/L. The growth of PANC-1 cells was inhibited by gemcitabine in a concentration-dependent manner (P < 0.0001) and the cell growth was also inhibited throughout the time course (P < 0.0001). The DNA fragmentation rate in the gemcitabine-treated group at 48 h was 44.7 %, whereas it was 25.3 % in the untreated group. The PAP mRNA expression was decreased after being treated with gemcitabine, whereas the TP53INP1 mRNA was increased by the gemcitabine treatment. Western blot analysis showed that phospho-GSK-3 beta(ser9) was induced by the gemcitabine treatment.CONCLUSION: Gemcitabine suppresses PANC-1 cell proliferation and induces apoptosis. Apoptosis is considered to be associated with the inhibition of PAP and GSK-3 beta, and the activation of TP53INP1 and pospho-GSK-3 beta(ser9). (C) 2006 The WJG Press. All rights reserved.