Pan Xue, Arumugam Thiruvengadam, Yamamoto Tameyoshi, Levin Pavel A, Ramachandran Vijaya, Ji Baoan, Lopez-Berestein Gabriel, Vivas-Mejia Pablo E, Sood Anil K, McConkey David J, Logsdon Craig D
Department of Gastroenterology, Changhai Hospital, Second Military Medical University, Shanghai, China.
Clin Cancer Res. 2008 Dec 15;14(24):8143-51. doi: 10.1158/1078-0432.CCR-08-1539.
Nuclear factor kappaB (NFkappaB) activity may increase survival and protect cancer cells from chemotherapy. Therefore, NFkappaB activity may be prognostic, and inhibition of NFkappaB may be useful for pancreatic cancer therapy. To test these hypotheses, we examined NFkappaB activity and the effects of inhibiting NFkappaB in several pancreatic cancer cell lines with differing sensitivities to gemcitabine.
The gemcitabine sensitivity of pancreatic cancer cell lines BxPC-3, L3.6pl, CFPAC-1, MPanc-96, PANC-1, and MIA PaCa-2 were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and fluorescence-activated cell sorting assays. NFkappaB levels were determined by electrophoretic mobility shift assay and reporter assays. The effects of gemcitabine on NFkappaB activity were determined in vitro and in vivo. NFkappaB was inhibited by silencing of the p65/relA subunit using small interfering RNA in vitro and by neutral liposomal delivery of small interfering RNA in vivo, and the effects were evaluated on gemcitabine sensitivity.
The cell lines L3.6pl, BxPC-3, and CFPAC-1 were sensitive, whereas MPanc-96, PANC-1, and MIA PaCa-2 were resistant to gemcitabine. No significant correlation was observed between basal NFkappaB activity and gemcitabine sensitivity. Gemcitabine treatment did not activate NFkappaB either in vitro or in vivo. Silencing of p65/relA induced apoptosis and increased gemcitabine killing of all gemcitabine-sensitive pancreatic cancer cells. No significant effects, however, were observed on gemcitabine-resistant pancreatic cancer cell lines either in vitro or in vivo.
NFkappaB activity did not correlate with sensitivity to gemcitabine. Silencing of p65/relA was effective alone and in combination with gemcitabine in gemcitabine-sensitive but not gemcitabine-resistant pancreatic cancer cells. Thus, NFkappaB may be a useful therapeutic target for a subset of pancreatic cancers.
核因子κB（NFκB）活性可能会提高生存率并保护癌细胞免受化疗影响。因此，NFκB活性可能具有预后价值，抑制NFκB可能对胰腺癌治疗有用。为验证这些假设，我们检测了NFκB活性以及在几种对吉西他滨敏感性不同的胰腺癌细胞系中抑制NFκB的效果。
通过3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐和荧光激活细胞分选测定法确定胰腺癌细胞系BxPC - 3、L3.6pl、CFPAC - 1、MPanc - 96、PANC - 1和MIA PaCa - 2对吉西他滨的敏感性。通过电泳迁移率变动分析和报告基因测定法确定NFκB水平。在体外和体内测定吉西他滨对NFκB活性的影响。在体外使用小干扰RNA沉默p65/relA亚基，并在体内通过中性脂质体递送小干扰RNA来抑制NFκB，然后评估其对吉西他滨敏感性的影响。
细胞系L3.6pl、BxPC - 3和CFPAC - 1对吉西他滨敏感，而MPanc - 96、PANC - 1和MIA PaCa - 2对吉西他滨耐药。未观察到基础NFκB活性与吉西他滨敏感性之间存在显著相关性。吉西他滨治疗在体外和体内均未激活NFκB。沉默p65/relA可诱导所有对吉西他滨敏感的胰腺癌细胞凋亡并增强吉西他滨的杀伤作用。然而，在体外和体内对吉西他滨耐药的胰腺癌细胞系均未观察到显著效果。
NFκB活性与对吉西他滨的敏感性无关。沉默p65/relA单独使用以及与吉西他滨联合使用时，对吉西他滨敏感而非耐药的胰腺癌细胞有效。因此，NFκB可能是一部分胰腺癌的有用治疗靶点。
