大黄素联合吉西他滨抑制体内外胰腺癌生长及其机制研究

Emodin Combined Gemcitabine Inhibited the Growth of Pancreatic Cancer in vitro and in vivo and Its Mechanisms Study

目的探讨大黄素联合吉西他滨对体内外胰腺癌生长的影响及其机制。方法采用吉西他滨(20μmol/L)、大黄素(40μmol/L)单独及联合作用胰腺癌细胞株SW1990后,CCK-8法检测细胞增殖;流式细胞术检测细胞凋亡;Westernblot检测Bax及Bcl-2蛋白表达。建立人胰腺癌裸鼠皮下移植瘤模型,分别给予大黄素、吉西他滨单独及联合用药,监测移植瘤体积变化;免疫组化法检测移植瘤组织中Ki-67、Bax及Bcl-2表达。结果与吉西他滨组及大黄素组比较,联合用药组显著降低SW1990细胞存活率,提高细胞凋亡率(P<0.05);与对照组比较,大黄素组及联合用药组明显上调SW1990细胞中Bax蛋白表达水平,抑制Bcl-2蛋白表达(P<0.05)。与对照组比较,大黄素组及联合用药组可显著抑制裸鼠胰腺癌皮下移植瘤生长,提高Bax在肿瘤组织中阳性表达,明显降低Ki-67和Bcl-2的阳性表达(P<0.05),以联合用药组作用最佳(P<0.05)。结论大黄素可能通过上调Bax表达和下调Bcl-2表达增强吉西他滨对体内外胰腺癌的抑制作用。

Objective To study the effect of emodin combined gemcitabine on the growth of pancreatic cancer in vivo and in vitro as well as its mechanisms. Methods After human pancreatic cancer cell line SW1990 was treated with emodin （40 μmol/L）, gemcitabine （20 μmol/L）, and emodin combined gemcitabine, the cell proliferation was detected by cell counting kit-8 （CCK-8） assay. The apoptosis of pancreatic cancer cells was detected using the flow cytometry （FCM）. The protein expressions of Bax and Bcl-2 were detected using Western blot. SW1990 cells were injected subcutaneously into nude mice to establish pancreatic xenograft tumors. The mice were then treated by emodin, gemcitabine, and emodin combined gemcitabine, respectively. The changes of tumor volume were monitored. The positive expressions of Ki-67, Bax, and Bcl-2 in the xenograff tumors were detected using immunohistochemical method. Results Emodin combined with gemcitabine induced a higher per-centage of growth inhibition and apoptosis in pancreatic cancer cell line SW1990 than that of gemcitabine or emo-din alone （P〈0.05）. The protein expression of Bax was up-regulated and that of Bcl-2 down-regulated in the emodin group and the emodin combined gemcitabine group when compared with the control group （ P 〈 0.05）. Emodin combined with gemcitabine could significantly inhibit the growth of pancreatic xenograft tumors, increase the positive expression of Bax in tumor tissues, obviously decrease the positive expressions of Ki-67 and Bcl-2 （ P 〈0. 05）. The optimal effects were obtained in the emodin combined gemcitabine group （ P 〈0. 05）. Conclusion Emodin could potentiate the inhibition of pancreatic cancer growth induced by gemcitabine both in vitro and in vivo, which might be achieved by up-regulating the expression of Bax and down-regulating the expression of Bcl-2.