CAJ | 학술논문

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Mia-PaCa-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&Pmpranolol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&Pmpranolol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VEGF mRNA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、105.32±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.
목적 탐토β수체재경전신경체질거갑신상선소(NE)유도인이선암세포주Mia-PaCa-2침습능력증강과정중적작용급기궤제.방법 역전록-취합매련반응(RT-PCR)법검측인이선암세포주β수체적표체.실험분위대조조、NE간예조、보내락이(Propranolol)간예조、보내락이화NE공동간예조(NE&Propranolol).채용Transwell침습실험검측세포침습능력적변화;RT-PCR법화면역세포화학법분별검측세포중기질금속단백매(MMP)-2、MMP-9、혈관내피생장인자(VEGF)mRNA화단백적표체.결과 인이선암세포주MiaPaCa-2화BxPC3균표체β1화β2수체.NE조、NE&Pmpranolol조、Propranolol조화대조조천막세포적흡광도치분별위0.78±0.02、0.32±0.03、0.26±0.01、0.28±0.02,NE조현저고우대조조화NE&Pmpranolol조(P<0.05),이Propranolol조천막세포수여대조조간차이무통계학의의(P>0.05);NE조중MMP-2、MMP-9화VEGF mRNA표체지수위0.87±0.02、1.04±0.02화0.92±0.01,단백표체회도치위131.20±2.34、105.32±7.21화115.60±5.03,현저고우대조조화NE&Propranolol조(P<0.05),Propranolol조여대조조간차이무통계학의의(P>0.05).결론 β수체재NE유도이선암세포침습능력증강적발전과정중발휘중요작용,NE통과β수체개도상조MMP-2、MMP-9화VEGF적표체진이증강이선암세포적침습능력.
Objective To investigate the effect and mechanism of β-adrenoceptors on norepi-nephrine-induced invasiveness of pancreatic cancer cell lines. Methods The expression of β-adrenocep-tors 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 VECF proteins were assayed by immunocytochemistry. Results Both MiaPaCa-2 and BxPC3 expressed β1-and β2-adrenocep-tors. 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 sig-nificant 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 pro-tein between propranolol 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-adre-noceptors.