ChRM3对胆管癌神经浸润的影响
|
摘要
目的探讨毒蕈碱型乙酰胆碱受体M3(ChRM3)对胆管癌神经浸润的影响。方法采用免疫组织化学方法检测60例胆管癌组织及30例正常胆管组织中ChRM3的表达水平;建立小鼠背根神经节(DRG)与胆管癌细胞RBE体外共培养模型,并加入ChRM3激动剂(匹罗卡品)及其拮抗剂(阿托品),通过观察神经纤维的生长和胆管癌RBE细胞对DRG和周围神经纤维的黏附及浸润,检测ChRM3在匹罗卡品和阿托品作用下对胆管癌RBE细胞浸润DRG及周围神经纤维的影响。结果 ChRM3在胆管癌组织中的阳性表达率高于正常胆管组织,差异有统计学意义(P<0.01)。神经浸润在胆管癌中发生率为91.7%(55/60)。DRG-RBE细胞共培养模型中加入匹罗卡品后,RBE细胞浸润DRG及周围神经纤维的细胞数明显增加,差异有统计学意义(P<0.05),这种作用在加入阿托品后明显减弱,差异有统计学意义(P<0.05)。而将阿托品单独作用于胆管癌组织,神经浸润结果与对照组比较无明显差异(P>0.05)。结论 ChRM3参与了胆管癌神经浸润过程,可能为抑制胆管癌的神经浸润提供了新治疗靶点。
Objective To explore the effect of muscarinic acetylcholine receptor M3( ChRM3) on the perineural invasion of cholangiocarcinoma( CLC). Methods Immunohistochemistry was used to detect the expression of ChRM3 in 60 CLCtissues and 30 normal bile duct tissues. A co-culture model of mouse dorsal root ganglia( DRG) and human CLC cell line RBE was established. The ChRM3 agonist pilocarpine and the ChRM3 antagonist atropine were added into the co-culture model to observe the growth of nerve fibers and the adhesion and invasion of RBE cells into the DRG and surrounding nerve fibers and to evaluate the effect of ChRM3 on the invasion of RBE cells into the DRG and surrounding nerve fibers. Results The positive rate of ChRM3 was significantly higher in the CLC tissues than in the normal bile duct tissues( P<0.01). The perineural invasion rate reached 91.7%( 55/60) in the CLC tissues. The number of RBE cells invading the DRG and surroundingnerve fibers was significantly increased after pilocarpine was added to the co-culture model( P<0.05). This effect was significantly weakened after atropine was added to the co-culture model( P<0.05). However,there was no significant change in the perineural invasion when atropine alone was added to the co-culture model( P> 0.05). Conclusion ChRM3 is involved in the perineural invasion of CLC cells,which may provide a new therapeutic target to inhibitthe perineural invasion of CLC.
Objective To explore the effect of muscarinic acetylcholine receptor M3( ChRM3) on the perineural invasion of cholangiocarcinoma( CLC). Methods Immunohistochemistry was used to detect the expression of ChRM3 in 60 CLCtissues and 30 normal bile duct tissues. A co-culture model of mouse dorsal root ganglia( DRG) and human CLC cell line RBE was established. The ChRM3 agonist pilocarpine and the ChRM3 antagonist atropine were added into the co-culture model to observe the growth of nerve fibers and the adhesion and invasion of RBE cells into the DRG and surrounding nerve fibers and to evaluate the effect of ChRM3 on the invasion of RBE cells into the DRG and surrounding nerve fibers. Results The positive rate of ChRM3 was significantly higher in the CLC tissues than in the normal bile duct tissues( P<0.01). The perineural invasion rate reached 91.7%( 55/60) in the CLC tissues. The number of RBE cells invading the DRG and surroundingnerve fibers was significantly increased after pilocarpine was added to the co-culture model( P<0.05). This effect was significantly weakened after atropine was added to the co-culture model( P<0.05). However,there was no significant change in the perineural invasion when atropine alone was added to the co-culture model( P> 0.05). Conclusion ChRM3 is involved in the perineural invasion of CLC cells,which may provide a new therapeutic target to inhibitthe perineural invasion of CLC.
引文
[1]RAZUMILAVA N,GORES G J.Cholangiocarcinoma[J].Lancet,2014,383(9935):2168-2179.
[2]FRIMAN S.Cholangiocarcinoma-current treatment options[J].Scand J Surg,2011,100(1):30-34.
[3]冯玉杰,张炳远,卢云.胆管癌周围神经浸润的研究进展[J].国际外科学杂志,2010,37(7):489-483.
[4]HASSAN M O,MAKSEM J.The prostatic perineural space and its relation to tumor spread:an ultrastructural study[J].Am J Surg Pathol,1980,4(2):143-148.
[5]BHUIYA M R,NIMURA Y,KAMIYA J,et al.Clinicopathologic studies on perineural invasion of bile duct carcinoma[J].Ann Surg,1992,215(4):344-349.
[6]刘磊,黄强,刘臣海,等.肝外胆管癌神经及脉管浸润转移的危险因素[J].肿瘤,2012,32(5):376-379.
[7]KANNO N,LESAGE G,PHINIZY J L,et al.Stimulation ofα2-adrenergic receptor inhibits cholangiocarcinoma growth through modulation of Raf-1 and B-Raf activities[J].Hepato-logy,2002,35(6):1329-1340.
[8]黄竹,刘子沛,夏锋,等.α1-肾上腺素受体对胆管癌细胞增殖的影响[J].中华消化外科杂志,2009,8(3):193-196.
[9]SHAH N,KHURANA S,CHENG K,et al.Muscarinic receptors and ligands in cancer[J].Am J Physiol Cell Physiol,2009,296(2):C221-232.
[10]SAID A H,HU S,ABUTALEB A,et al.Interacting post-muscarinic receptor signaling pathways potentiate matrix metalloproteinase-1 expression and invasion of human colon cancer cells[J].Biochemical Journal,2017,474(5):647-665.
[11]TROMBINO S,CESARIO A,MARGARITORA S,et al.α7-Nicotinic acetylcholine receptors affect growth regulation of human mesothelioma cells role of mitogen-activated protein kinase pathway[J].Cancer Res,2004,64(1):135-139.
[12]SONG P,SEKHON H S,PROSKOCIL B,et al.Synthesis of acetylcholine by lung cancer[J].Life Sci,2003,72(18):2159-2168.
[13]FRUCHT H,JENSEN R T,DEXTER D,et al.Human colon cancer cell proliferation mediated by the M3 muscarinic cholinergic receptor[J].Clin Cancer Res,1999,5(9):2532-2539.
[14]WEGENER C,HAMASAKA Y,NASSEL D R.Acetylcholine increases intracellular Ca2+via nicotinic receptors in cultured PDF-containing clock neurons of Drosophila[J].J Neurophysiol,2004,91(2):912-922.
[15]张亮,宦宏波,温旭东,等.毒蕈碱型胆碱能受体M3调控肝癌细胞侵袭转移的研究[J].第三军医大学学报,2017,39(6):509-514.
[16]ELSING C,HUBNER C,FITSCHER B A,et al.Muscarinic acetylcholine receptor stimulation of biliary epithelial cells and its effect on bile secretion in the isolated perfused liver[J].Hepatology,1997,25(4):804-814.
[17]FENG Y J,ZHANG B Y,YAO R Y,et al.Muscarinic acetylcholine receptor M3 in proliferation and perineural invasion of cholangiocarcinoma cells[J].Hepatobiliary Pancreat Dis Int,2012,11(4):418-423.
[18]刘昆鹏,张炳远,卢云,等.Transwell侵袭实验测定M受体对胆管癌细胞侵袭的影响[J].国际外科学杂志,2011,38(5):298-301.
[19]RADU A,PICHON C,CAMPARO P,et al.Expression of follicle-stimulating hormone receptor in tumor blood vessels[J].N Engl JMed,2010,186(2):1621-1630.
[2]FRIMAN S.Cholangiocarcinoma-current treatment options[J].Scand J Surg,2011,100(1):30-34.
[3]冯玉杰,张炳远,卢云.胆管癌周围神经浸润的研究进展[J].国际外科学杂志,2010,37(7):489-483.
[4]HASSAN M O,MAKSEM J.The prostatic perineural space and its relation to tumor spread:an ultrastructural study[J].Am J Surg Pathol,1980,4(2):143-148.
[5]BHUIYA M R,NIMURA Y,KAMIYA J,et al.Clinicopathologic studies on perineural invasion of bile duct carcinoma[J].Ann Surg,1992,215(4):344-349.
[6]刘磊,黄强,刘臣海,等.肝外胆管癌神经及脉管浸润转移的危险因素[J].肿瘤,2012,32(5):376-379.
[7]KANNO N,LESAGE G,PHINIZY J L,et al.Stimulation ofα2-adrenergic receptor inhibits cholangiocarcinoma growth through modulation of Raf-1 and B-Raf activities[J].Hepato-logy,2002,35(6):1329-1340.
[8]黄竹,刘子沛,夏锋,等.α1-肾上腺素受体对胆管癌细胞增殖的影响[J].中华消化外科杂志,2009,8(3):193-196.
[9]SHAH N,KHURANA S,CHENG K,et al.Muscarinic receptors and ligands in cancer[J].Am J Physiol Cell Physiol,2009,296(2):C221-232.
[10]SAID A H,HU S,ABUTALEB A,et al.Interacting post-muscarinic receptor signaling pathways potentiate matrix metalloproteinase-1 expression and invasion of human colon cancer cells[J].Biochemical Journal,2017,474(5):647-665.
[11]TROMBINO S,CESARIO A,MARGARITORA S,et al.α7-Nicotinic acetylcholine receptors affect growth regulation of human mesothelioma cells role of mitogen-activated protein kinase pathway[J].Cancer Res,2004,64(1):135-139.
[12]SONG P,SEKHON H S,PROSKOCIL B,et al.Synthesis of acetylcholine by lung cancer[J].Life Sci,2003,72(18):2159-2168.
[13]FRUCHT H,JENSEN R T,DEXTER D,et al.Human colon cancer cell proliferation mediated by the M3 muscarinic cholinergic receptor[J].Clin Cancer Res,1999,5(9):2532-2539.
[14]WEGENER C,HAMASAKA Y,NASSEL D R.Acetylcholine increases intracellular Ca2+via nicotinic receptors in cultured PDF-containing clock neurons of Drosophila[J].J Neurophysiol,2004,91(2):912-922.
[15]张亮,宦宏波,温旭东,等.毒蕈碱型胆碱能受体M3调控肝癌细胞侵袭转移的研究[J].第三军医大学学报,2017,39(6):509-514.
[16]ELSING C,HUBNER C,FITSCHER B A,et al.Muscarinic acetylcholine receptor stimulation of biliary epithelial cells and its effect on bile secretion in the isolated perfused liver[J].Hepatology,1997,25(4):804-814.
[17]FENG Y J,ZHANG B Y,YAO R Y,et al.Muscarinic acetylcholine receptor M3 in proliferation and perineural invasion of cholangiocarcinoma cells[J].Hepatobiliary Pancreat Dis Int,2012,11(4):418-423.
[18]刘昆鹏,张炳远,卢云,等.Transwell侵袭实验测定M受体对胆管癌细胞侵袭的影响[J].国际外科学杂志,2011,38(5):298-301.
[19]RADU A,PICHON C,CAMPARO P,et al.Expression of follicle-stimulating hormone receptor in tumor blood vessels[J].N Engl JMed,2010,186(2):1621-1630.