目的: 探讨乌苏酸联合吉西他滨对胰腺癌PANC-1细胞增殖和凋亡的影响。 方法: 以乌苏酸、吉西他滨单独用药作 用于体外培养的胰腺癌细胞PANC-1细胞，采用细MTT法检测细胞半抑制浓度（IC 50 ），确定药物给药浓度；以乌苏酸(2 μ mol/L) 、 吉西他滨(0.2 μ mol/L)单独及联合作用于PANC-1细胞，采用锥虫蓝染色法检测PANC-1细胞存活率，PI染色后PANC-1细胞中检 测细胞凋亡，采用细胞划痕实验检测PANC-1细胞迁移和伤口愈合情况，采用Western blotting检测对照组、乌苏酸组与吉西他滨 组单独及联合用药组细胞中P-JNK、Bcl-2、IL-6、P-Stat 3、NF-κB和Cox-2蛋白的表达。 结果: 乌苏酸和吉西他滨对胰腺癌 PANC-1细胞均有较强的抑制作用，其IC 50 分别是13.67和2.78 μ mol/L，据此选择它们的实验浓度分别为2和0.2 μ mol/L。两者联 合用药比分别单独用药能更显著抑制癌细胞的增殖活性[(46.47±5.07)% vs (78.38±8.65)%、(76.12±3.23)%，均P<0.05]，能更明显 诱导癌细胞凋亡[(39.78±7.01)% vs (20.35±8.51)%、(20.35±8.51)%，均P<0.01]和抑制细胞迁移能力（P<0.01），同时更显著抑制凋亡 相关蛋白Bcl-2、IL-6的表达及促进P-JNK的表达。 结论: 乌苏酸和吉西他滨协同增强抑制胰腺癌PANC-1细胞的增殖和迁移以 及促凋亡作用，其机制可能与抑制Bcl-2、IL-6、P-Stat 3和促进P-JNK蛋白表达有关。

Objective: To study the effects of ursolic acid cooperated with gemcitabine on proliferation and apoptosis of pancreatic can- cer PANC-1 cells. Methods: Human pancreatic cancer cell line PANC-1 was cultured in vitro with ursolic acid and gemcitabine respec- tively; and MTT assay was used to determine the IC 50 of ursolic acid and gemcitabine, thus obtaining the best drug concentration. Urso- lic acid (2 μ mol/L) and gemcitabine (0.2 μ mol/L) alone or in combination was used to treat PANC-1 cells; trypan blue assay was used to test cell viability, and PI staining was used to examine the cell apoptosis; wound healing was used to detect the proliferation and mi- gration of PANC-1 cells. The protein expressions of P-JNK, Bcl-2, IL-6, P-Stat 3, NF-κB and Cox-2 in cells of each treatment group were detected using Western blotting. Results: Both ursolic acid and gemcitabine could significantly inhibit the proliferation of PANC-1 cells, and the IC 50 is 13.67 and 2.78 μ mol/L, respectively; and the final concentrations were determined at 2 and 0.2 μ mol/L for ursolic acid and gemcitabine, respectively. Compared with single drug treatment, the combined treatment exerted a more prominent cell proliferation inhibition effect ([46.47±5.07]% vs [78.38±8.65]%, [76.12±3.23]%, all P<0.05), apoptosis-induction effect ([39.78± 7.01]% vs [20.35±8.51]%, [20.35±8.51]%, all P<0.01) and migration inhibition effect (P<0.01) on PANC-1 cells. Western blotting showed that the combined treatment strongly inhibited Bcl-2 and IL-6 expression, accelerated P-JNK protein expression compared with single drug treatment. Conclusion: The synergistic effect of ursolic acid and gemcitabine enhanced the inhibition on proliferation, mi- gration, and promoted cell apoptosis of human pancreatic cancer cell line PANC-1, the mechanism may be associated with inhibition of Bcl-2, Il-6, P-stat 3 proteins and promotion of P-JNK protein.

国家自然科学基金资助项目（No. 81272452）