摘要
目的观察瑞芬太尼对胰腺癌BxPC-3细胞增殖、凋亡的影响并探讨其作用机制。方法在体外培养BxPC-3细胞,用不同浓度瑞芬太尼(终浓度0、0.5、1、2、4、8 mg/mL)分别处理24、48、72 h,采用CCK-8比色法检测药物处理后细胞活力;瑞芬太尼(0、1、2、4 mg/mL)处理BxPC-3细胞48 h后,采用PI/Annexin V双染法检测细胞凋亡率;采用Western blot法检测药物对细胞Bax、Bcl-2、t-Akt、p-Akt及p-GSK-3β蛋白表达的影响;采用端粒酶重复序列扩增法检测端粒酶活性。结果 CCK-8比色法结果显示,瑞芬太尼处理后可抑制BxPC-3细胞的增殖,且呈剂量、时间依赖性;瑞芬太尼处理后,PI/Annexin V双染后流式细胞仪检测结果显示,随着药物浓度的增加,BxPC-3细胞凋亡率也逐渐增加(P<0.05);Western blot结果显示,随着药物浓度的增加,抗凋亡蛋白Bcl-2表达量逐渐降低,且促凋亡蛋白Bax的表达量逐渐增高,BxPC-3细胞Akt总量未发生变化,而p-Akt及其下游蛋白p-GSK-3β的表达逐渐降低(P<0.05);端粒酶重复序列扩增法检测结果显示,随着瑞芬太尼浓度的增加,BxPC-3细胞端粒酶活性降低(P<0.05)。结论瑞芬太尼在体外能抑制胰腺癌BxPC-3细胞增殖并促进其凋亡,其作用机制可能与抑制Akt通路,上调Bax表达量,降低Bcl-2表达量,进而减弱端粒酶活性有关。
Objective To observe the effect of remifentanil on cell proliferation and apoptosis in pancreatic cancer cellline BxPC-3 cells and explore the possible mechanisms. Methods BxPC-3 cells were cultured in vitro. After treatment byremifentanil at different concentrations（0, 0.5, 1, 2, 4, 8 mg/mL）respectively at different time（24, 48, 72 h）, the cell viabilitywas determined by the CCK-8 method. Apoptosis was analyzed by PI/Annexin V flow cytometry. Western blot were used forBcl-2, Bax, Akt, p-Akt and p-GSK-3β protein analysis. Telomerase activity was analyzed by telomerase repeat amplificationprotocol（TRAP）. Results From the data of CCK-8, the cell proliferation of BxPC-3 cells was inhibited by remifentanil in adose-dependent and time-dependent manner（P〈0.05）. After treated with remifentanil, PI/Annexin V flow cytometry showedthe apoptosis rate of BxPC-3 cells increased（P〈0.05）. Western blot showed the treatment with remifentanil promoted theexpression of pro-apoptotic factor Bax, and suppressed the expression of anti-apoptotic factor Bcl-2（P〈0.05）. And theexpressions of p-Akt and p-GSK-3β were suppressed. TRAP showed the telomerase activity was decreased（P〈0.05）.Conclusion Remifentanil inhibits the cell proliferation and induces cell apoptosis in BxPC-3 cells, and the effects ofantitumor may be associated with promotion the expression of pro-apoptotic factor Bax, inhibition of the expression of Akt andBcl-2, thus inhibiting the telomerase activity.
出处
《中国热带医学》
CAS
2017年第12期1193-1197,共5页
China Tropical Medicine
