AG490联合健择对人胰腺癌细胞生长的影响

Effect of AG490 combined with gemcitabine on the growth of human pancreatic cancer cells

目的:探讨Janus激酶(JAK)特异性抑制剂AG490联合化疗药物健择对人胰腺癌细胞系SW1990的生长增殖及STAT3转导通路的影响和其机制.方法:人胰腺癌细胞系SW1990分为对照组、AG490组、健择组及AG490+健择处理组.培养48 h后,MTT法检测细胞增殖状态,流式细胞仪检测细胞凋亡.Western blot和RT-PCR检测STAT3,Cyclin D1,Bcl-xL,Bax,Survivin的表达情况.结果:AG490组和健择组的细胞增殖明显低于对照组(2.20±0.25,2.30±0.220 vs 3.78±0.42,P<0.05),凋亡率明显高于对照组(35.40%±3.08%,34.64%±1.38% vs 16.49%±1.45%,P<0.05)。并且,AG490+健择组细胞增殖(1.49±0.15)明显低于明显低于AG490或健择组(P<0.05),而凋亡率(43.80%±1.57%)则明显高于AG490或健择组(P<0.05).AG490处理SW1990 48 h后,p-STAT3表达明显低于对照组(13.83%±0.64% vs 79.87%±1.43%,P<0.05),同时Cyclin D1(mRNA:15.63%±0.59% vs 43.83%±0.64%,P<0.05:蛋白:17.50%±0.92% vs 49.87%±1.27%,P<0.05),Bcl-xL(mRNA:13.93%±0.21% vs 75.70%±0.46%,P<0.05:蛋白:34.17%±1.70% vs 83.93%±0.80%,P<0.05)和Survivin(mRNA:58.27%±0.42% vs 82.93%±1.68%,P<0.05:蛋白:13.23%±1.03% vs 18.60±1.08%,P<0.05)表达也明显降低,而Bax的表达则明显增高(mRNA:10.33%±1.18% vs 5.43%±0.70%,P<0.05:蛋白:13.07%±1.04% vs 6.23%±2.40%,P<0.05),健择处理组上述指标与对照组相似.结论:阻断STAT3信号转导通路可以抑制人胰腺癌细胞增殖,促进其凋亡,健择联合AG490能起协同作用.AG490联合健择可能为胰腺癌治疗提供新的思路.

AIM： To investigate the effect of Janus kinase （JAK） specific inhibitor AG490 combined with gemcitabine on the proliferation of human pancreatic cancer cell line SW1990 and STAT3 signal transduction pathway as well as their mechanisms.METHODS： Human pancreatic cancer cell line SW1990 was divided into control group, AG490- treated group, gemcitabine-treated group and AG490 ＋ gemcitabine-treated group. After 48 hours, the proliferation of SW1990 cells was detected by MTT assay. Flow cytometry was used to examine cell apoptosis. The expression of STAT3, Cyclin D1, Bcl-xL and Bax were detected by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot, respectively. RESULTS： The proliferation of SW1990 cells was significantly lower in AG490 or gemcitabine group than that in control group （2.20 ± 0.25, 2.30 ± 0.220 vs 3.78 ± 0.42, P 〈 0.05）, but the apoptosis rate was markedly higher （35.40% ± 3.08%, 34.64% ± 1.38% vs 16.49% ± 1.45%, P 〈0.05）. Moreover, the proliferation （1.49 ± 0.15） and apoptosis （43.80% ± 1.57%） had notable difference between AG490＋gemcitabine group and AG490 or gemcitabine group. After 48 hours, AG490 remarkably down-regulated the expression of p-STAT3 （13.83% ± 0.64% vs 79.87% ±1.43%, P 〈 0.05）, and the expression of Cyclin D1 （mRNA： 15.63% ± 0.59% vs 43.83% ± 0.64%, P 〈 0.05; protein： 17.50%±0.92% vs 49.87% ±1.27%, P 〈 0.05）, Bcl-xL （mRNA： 13.93% ± 0.21% vs 75.70% ± 0.46%, P 〈 0.05; protein： 34.17% ± 1.70% vs 83.93% ± 0.80%, P 〈 0.05） and Survivin （mRNA： 58.27% ± 0.42% vs 82.93% ± 1.68%, P 〈 0.05; protein： 13.23%±1.03% vs 18.60 ± 1.08%, P 〈 0.05） were also decreased in comparison with that in control group; however, Bax expression was increased （mRNA： 10.33% ±1.18% vs 5.43% ±0.70%, P 〈 0.05; protein： 13.07% ± 1.04% vs 6.23% ± 2.40%, P 〈 0.05）. There was no difference between gemcitabine and control group. CONCLUSION： Blockade of STAT3 signal pathway inhibits the proliferation and promotes the apoptosis of SW1990 cells. AG490 combined with gemcitabine shows a synergic effect, which provides a new therapeutic approach for pancreatic cancer therapy.