舒芬太尼对BT474细胞恶性生物学行为影响及机制研究

The Influence and Mechanism of Sufentanil on Malignant Biological Behavior of BT474 Cells

目的探讨舒芬太尼对BT474人乳腺导管癌细胞恶性生物学行为的影响及作用机制。方法体外培养BT474人乳腺导管癌细胞,随机将细胞分为对照组(C组)和舒芬太尼组,C组仅使用1640完全型培养基培养,舒芬太尼组分别给予含不同剂量舒芬太尼[0.1 nmol·L^(-1)(L组),1 nmol·L^(-1)(M组),10 nmol·L^(-1) L(H组)]的1640完全型培养基培养;处理时间24 h。采用平板克隆形成实验检测细胞克隆形成能力,划痕实验检测细胞迁移能力,Transwell侵袭实验检测细胞侵袭能力,血管形成实验检测血管生成能力,Western blot实验检测细胞生长、转移和上皮-间质转化(epithelial-mesenchymal transition,EMT)相关蛋白的表达。结果与C组比较,舒芬太尼组细胞克隆形成能力逐渐降低、细胞迁移率和细胞侵袭数逐渐减少(P均<0.05)、血管生成能力逐渐降低且均与浓度呈剂量相关性(P均<0.05);Western blot结果显示,与C组比较,L组MMP-9、E-cadherin、N-cadherin蛋白表达水平差异均无统计学意义(P均>0.05),M组和H组MMP-9、N-cadherin蛋白表达量下调,E-cadherin蛋白表达量上调(P均<0.05),VEGF与Vimentin蛋白随舒芬太尼组干预剂量浓度的增加,蛋白表达均呈下调改变(P均<0.05)。结论舒芬太尼可抑制BT474人乳腺导管癌细胞生长、转移及血管形成,这一过程可能与EMT的发生有关。

Objective To explore the influence and mechanism of sufentanil on the malignant biological behavior of BT474 human breast catheter cancer cells.Methods BT474 human breast catheter cancer cells were cultured in vitro and randomly divided into control group(C group)and sufentanil group,C Group was only cultured in 1640 complete medium,the intervention dose of sufentanil group was 0.1 nmol·L^(-1)(L group),1 nmol·L^(-1)(M group),10 nmol·L^(-1)(H group),processing time was 24 h.Flat clone formation experiment was used to detect cell cloning capacity.Scratch experiments were used to detect cell migration ability.Transwell invasion experiments were used to detect cell invasion ability.Vascular formation experiment was used to detect the angiogenesis capacity.Western Blot experiment was used to detect the expression of proteins associated with cell growth,metastasis,and epithelial-mesenchymal transition(EMT).Results Compared with C group,the cell clone formation ability of sufentanil group gradually decreased,the cell migration rate and the number of cell invasion were gradually decreased(P all<0.05),and the angiogenesis ability gradually decreased and all were dose-related with concentration(P all<0.05).Western blot experiments showed that compared with C group,the protein expression levels of MMP-9,E-cadherin and N-cadherin in L group had no significant effect(P all>0.05).The expression of MMP-9 and N-cadherin protein was down-regulated in M and H group,and the expression of E-cadherin protein was up-regulated(P all<0.05);VEGF and Vimentin protein expressions were down-regulated with the increase of the intervention dose of sufentanil group(P all<0.05).Conclusion Sufentanil can inhibit the growth,metastasis and angiogenesis of BT474 human breast catheter cancer cells ductal carcinoma cells.This process may be related to the occurrence of EMT.