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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 the proliferation and apoptosis of pancreatic cancer BxPC-3 cells and to explore its mechanism of action. Methods BxPC-3 cells were cultured in vitro and treated with different concentrations of remifentanil (final concentrations of 0, 0.5, 1, 2, 4, and 8 mg/mL) for 24, 48, and 72 hours, respectively, using the CCK-8 colorimetric method. Cell viability was measured after drug treatment; BxPC-3 cells were treated with remifentanil (0, 1, 2, 4 mg/mL) for 48 h, and the rate of apoptosis was measured by PI/Annexin V double staining; Western blot was used. The effect of drug on the expression of Bax, Bcl-2, t-Akt, p-Akt and p-GSK-3β protein was detected. The telomerase activity was detected by telomerase repeat amplification. Results CCK-8 colorimetric results showed that remifentanil treatment inhibited the proliferation of BxPC-3 cells in a dose- and time-dependent manner; after remifentanil treatment, PI/Annexin V double-stained flow cells The assay results showed that with the increase of drug concentration, the apoptosis rate of BxPC-3 cells also gradually increased (P<0.05); Western blot results showed that with the increase of drug concentration, the expression of anti-apoptotic protein Bcl-2 gradually The expression of pro-apoptotic protein Bax increased gradually and the total amount of Akt in BxPC-3 cells did not change, while the expression of p-Akt and its downstream protein p-GSK-3β decreased gradually (P<0.05); The enzyme repeat amplification assay results showed that with the increase of remifentanil concentration, the telomerase activity of BxPC-3 cells was decreased (P<0.05). Conclusion Remifentanil can inhibit the proliferation and promote apoptosis of pancreatic cancer BxPC-3 cells in vitro. Its mechanism may be related to inhibition of Akt pathway, up-regulation of Bax expression, decrease of Bcl-2 expression, and thus decrease of telomerase activity. .