摘要
目的探讨β受体在经典神经递质去甲肾上腺素（NE）诱导人胰腺癌细胞株MiaPaCa-2侵袭能力增强过程中的作用及其机制。方法逆转录-聚合酶链反应（RT-PCR）法检测人胰腺癌细胞株β受体的表达。实验分为对照组、NE干预组、普萘洛尔（Propranolol）干预组、普萘洛尔和NE共同干预组（NE＆Propranolol）。采用Transwell侵袭实验检测细胞侵袭能力的变化；RT—PCR法和免疫细胞化学法分别检测细胞中基质金属蛋白酶（MMP）-2、MMP-9、血管内皮生长因子（VEGF）mRNA和蛋白的表达。结果人胰腺癌细胞株MiaPaCa-2和BxPC3均表达β1和β2受体。NE组、NE＆Propranolol组、Propranolol组和对照组穿膜细胞的吸光度值分别为0．78±0．02、0．32±0．03、0．26±0．01、0．28±0．02，NE组显著高于对照组和NE＆Propranolol组（P〈0．05），而Propranolol组穿膜细胞数与对照组间差异无统计学意义（P〉0．05）；NE组中MMP．2、MMP-9和VEGFmRNA表达指数为0．87±0．02、1．04±0．02和0．92±0．01，蛋白表达灰度值为131．20±2．34、105．324-7．21和115．60±5．03，显著高于对照组和NE＆Propranolol组（P〈0．05），Propranolol组与对照组间差异无统计学意义（P〉0．05）。结论β受体在NE诱导胰腺癌细胞侵袭能力增强的发展过程中发挥重要作用，NE通过β受体介导上调MMP-2、MMP-9和VEGF的表达进而增强胰腺癌细胞的侵袭能力。
Objective To investigate the effect and mechanism of β-adrenoceptors on norepinephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenoceptors mRNA in human pancreatic cancer cell lines MiaPaCa-2 and BxPC3 was detected by using RT-PCR.The cells were randomly divided into control group, 10 mol/L NE intervention group, 1 mol/L propranolol intervention group and NE ＋ propranolol intervention group. After 48 h, transwell invasiveness test was used to examine the changes in invasive ability of MiaPaCa-2. The expression of MMP-2, MMP-9 and VEGF mRNA was measured by semi-quantitative RT-PCR. The levels of MMP-2, MMP-9 and VEGF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenoceptors. The absorbance （A） values of invasive cells in NE, NE ＋ propranolol, propranolol and control groups were 0.78 ± 0.02,0.32 ± 0.03,0.26 ± 0.01 and 0.28 ± 0.02, respectively, and those in NE intervention group were significantly higher than in control and NE ＋ propranolol groups （P 〈 0.05 ）. There was no significant difference in the number of invasive cells between propranolol and control groups （ P 〉 0.05 ）. In NE group, the expression index of MMP-2, MMP-9 and VEGF mRNA was 0.87 ±0. 02,1.04 ± 0.02 and 0.92 ± 0.01, and the gray value of the protein expression was 131.20 ± 2.34,105.32 ± 7.21 and 115.60 ± 5.03, respectively, which were higher than those in control and NE ＋ propranolol groups （ P 〈 0.05 ）.There was no significant difference in the expression levels of MMP-2, MMP-9 and VEGF mRNA and protein between propranolot and control groups （P 〉 0.05）. Conclusion β-adrenoceptors play an important role in the process of norepinephrine-induced invasiveness of pancreatic cancer cells. NE can promote the invasiveness of MiaPaCa-2 through up-regulating the expression of MMP-2, MMP-9 and VEGF via β-adrenoeeptors.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2009年第7期827-829,共3页
Chinese Journal of Experimental Surgery
