NF-κB在大黄素联合吉西他滨治疗裸鼠胰腺癌肝转移中的作用

Role of NF-κB in Anti-Tumor Effect of Emodin Combined Gemcitabine on Hepatic Metastasis of Human Pancreatic Carcinoma in Nude Mice

目的:观察大黄素联合吉西他滨对裸鼠胰腺癌肝转移模型的影响及其可能机制。方法:建立裸鼠胰腺癌肝转移模型,随机(随机数字法)分为对照组、大黄素组、吉西他滨组和联合用药组(n=10),观察各组荷瘤裸鼠肝转移瘤生长情况;免疫组织化学法检测肿瘤组织中Ki-67和CD-34的表达;Tunel法检测肿瘤组织细胞凋亡;凝胶电泳迁移率实验(EMSA)检测肿瘤细胞中NF-κB活性;Western blotting法检测肿瘤组织中Survivin、Caspase-3、VEGF和MMP-9的表达。结果:相对吉西他滨组,吉西他滨联合大黄素可显著抑制胰腺癌肝转移瘤生长;与吉西他滨组相比较,大黄素单独或联合吉西他滨可抑制体内胰腺癌肝转移瘤中Ki-67和CD34的阳性表达,并降低NF-κB活性以及Survivin、VEGF和MMP-9的表达水平,同时上调Caspase-3的表达。结论:大黄素可增强吉西他滨对体内外胰腺癌肝转移瘤的增殖抑制和诱导凋亡作用,该作用可能通过下调NF-κB及其调控蛋白而实现。

Objective：To investigate the anti- tumor effect of emodin combined gemcitabine on the model of hepatic metastasis of human pancreatic carcinoma in nude mice. Methods：After model of hepatic metastasis of human pancreatic carcinoma was established,the models were randomized（ random number） into 4 groups（ n = 10） for experiments： the control group,the gemcitabine group,the emodin group and the combination group. The hepatic metastatic foci of pancreatic tumor weight and inhibition rate were evaluated respectively after the mice were sacrificed. Immunohistochemistry（IHC） was used to detect the positive expressions of Ki-67 and CD34 in the tumors. The Tunel test kit was used to explore cellular apoptosis in tumor tissue. The activity of NF- κB in pancreatic cancer cells was measured by electrophoretic mobility shift assay（EMSA）. Expressions of Survivin,Caspase- 3,VEGF and MMP- 9 were detected by Western blotting. Results：Administration of gemcitabine or emodin significantly decreased tumor weight compared to untreated control.Furthermore,the combination of emodin and gemcitabine resulted in a substantial delay of tumor growth as compared to either agent alone. The positive expressions of Ki- 67 and CD34 in the tumors of combination group were significantly lower than those in control group. Apoptosis occurred more frequently in the combination group than in the control. EMSA and Western blot analysis demonstrated that the combination therapy significantly down- regulated NF- κB,Survivin,VEGF and MMP- 9 in pancreatic cancer cells. Conclusion：The combination of emodion and gemcitabine resulted in the inhibition of hepatic metastasis of human pancreatic carcinoma in nude mice,which partially through down- regulation of NF-κB and its regulated proteins.