GPC1在胆道闭锁患儿肝组织中表达水平及临床意义的初步探索
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摘要
目的初步探讨GPC1在胆道闭锁患儿发病过程中的临床意义。方法选取深圳市儿童医院2016年7月至2018年2月收治的胆道闭锁患儿35例作为观察组,同期选取本院收治的先天性胆总管扩张症患儿42例作为对照组。通过免疫组化结果了解GPC1在胆道闭锁肝脏中的分布情况;通过qPCR及Western Blot对胆道闭锁(BA)及先天性胆总管扩张症(CC)患儿肝脏中的CGPC1蛋白水平、GPC1 mRNA水平进行检测。结果本研究结果显示GPC1主要分布在胆道闭锁肝脏胆管细胞顶部,观察组和对照组患儿GPC1 mRNA表达水平(0.65±0.05 vs. 2.54±0. 12,P<0.05)、蛋白表达量(1.31±0.14 vs. 2.83±0.25,P<0.05)比较,差异有统计学意义。结论 GPC1 mRNA和蛋白产物的低表达可能与胆道闭锁的发病密切相关。
Objective To explore the clinical significance of GPC1 in the pathogenesis of children with biliary atresia(BA). Methods A total of 35 hospitalized BA children at Children s Hospital of Shenzhen from July 2016 to February 2018 were recruited into observation group. And another 42 hospitalized children with congenital choledochal dilatation were selected as control group. The distribution of GPC1 protein in observation group was detected by immunohistochemistry and the expression levels of GPC1 and protein expression in liver samples of two groups were detected by quantitative polymerase chain reaction(qPCR) and Western blot. Results GPC1 was distributed predominantly at the top of cholangiocarcinoma cells. The expression of GPC1 mRNA in observation and control groups(0.65 ±0.05 vs. 2.54 ±0. 12,P< 0.05) and protein expression(1.31±0. 14 vs. 2. 83 ±0. 25,P <0.05) showed statistical differences. Conclusion The expression of GPCI may be closely correlated with the pathogenesis of BA.
Objective To explore the clinical significance of GPC1 in the pathogenesis of children with biliary atresia(BA). Methods A total of 35 hospitalized BA children at Children s Hospital of Shenzhen from July 2016 to February 2018 were recruited into observation group. And another 42 hospitalized children with congenital choledochal dilatation were selected as control group. The distribution of GPC1 protein in observation group was detected by immunohistochemistry and the expression levels of GPC1 and protein expression in liver samples of two groups were detected by quantitative polymerase chain reaction(qPCR) and Western blot. Results GPC1 was distributed predominantly at the top of cholangiocarcinoma cells. The expression of GPC1 mRNA in observation and control groups(0.65 ±0.05 vs. 2.54 ±0. 12,P< 0.05) and protein expression(1.31±0. 14 vs. 2. 83 ±0. 25,P <0.05) showed statistical differences. Conclusion The expression of GPCI may be closely correlated with the pathogenesis of BA.
引文
1 Asai A,Miethke A,Bezerra JA. Pathogenesis of biliary atresia:defining biology to understand clinical phenotypes[D].Nat Rev Gastroenterol Hepatol,2015,12(6):342-345. DOI:10.1038/nrgastro. 2015, 74.
2 Wen J, Xiao Y, Wang J, et al. Low doses of CMV induce autoimmune-mediated and inflammatory responses in bile duct epithelia of regulatory T cell-depleted neonatal mice[J]. Lab Invest,2015, 95(2):180-192. DOI:10. 1038/labinvest.2014.148.
3郭振亚,张若岩,柳明江,等.先天性胆道闭锁的病因研究进展[J].临床肝胆病杂志,2015,31(8):1343-1346.DOI:10. 3969/j. issn. 1001-5256.2015.08. 043.Guo ZY, Zhang RY, Liu MG, et al. Etiology of congenital biliary atresia[J]. Journal of Clinical Hepatobiliary Diseases,2015,31(8):1343-1346. DOI:10. 3969/j. issn. 1001-5256.2015.08.043.
4 Tang V,Cofer ZC,Cui S, et al. Loss of a candidate biliary atresia susceptibility gene,add3 a,causes biliary developmental defects in zebrafish[J]. J Pediatr Gastroenterol Nutr,2016,63(5):524-530. DOI:10. 1097/MPG. 0000000000001375.
5李智涵,王斌.Ⅲ型胆道闭锁手术治疗的研究进展[J].临床小儿外科杂志,2016,15(1):11-13. DOI:10.3969/j.issn. 1671-6353.2016.01.004. Li ZH,Wang B. Advances in surgical treatment of biliary atresia of typeⅢ[J]. J Clin Ped Sur,2016,15(1):11-13. DOI:10. 3969/j. issn. 1671-6353.2016.01.004.
6 Asai A, Miethke A, Bezerra JA. Pathogenesis of biliary atresia:defining biology to understand clinical phenotypes[J].Nat Rev Gastroenterol Hepatol,2015,12(6):342-352. DOI:10. 1038/nrgastro. 2015. 74.
7 Hill JJ, Tremblay TL,Fauteux F,et al. Glycoproteomic comparison of clinical triple-negative and luminal breast tumors[J]. Journal of Proteome Research,2015,14(3):1376-1388. DOI:10. 1021/pr500987r.
8 Sadek KH,Ezzat S, Abdel-Aziz SA,et al. Macrophage migration inhibitory factor(MIF)gene promotor polymorphism is associated with increased fibrosis in biliary atresia patients,but not with disease susceptibility[J]. Ann Hum Genet,2017,81(5):177-183.DOI:10. 1111/ahg. 12199.
9 Tang V,Cofer ZC,Cui S,et al. Loss of a candidate biliary atresia susceptibility gene, add3 a, causes biliary developmental defects in zebrafish[J]. J Pediatr Gastroenterol Nutr,2016,63(5):524-530. DOI:10. 1097/MPG. 0000000000001375.
10 Leyva-Vega M, Gerfen J, Thiel BD,et al. Genomic alterations in biliary atresia suggest region of potential disease susceptibility in 2q37. 3[J]. Am J. Med. Genet. A, 2010,152A(4):886-895. DOI:10.1002/ajmg. a. 33332.
11 Cui S,Leyva-Vega M,Tsai EA,et al. Evidence from human and zebrafish that GPC1 is a biliary atresia susceptibility gene[J]. Gastroenterology,2013,144(5):1107-1115.DOI:10. 1053/j. gastro. 2013.01.022.
12 Ke J,Zeng S,Mao J,et al. Common genetic variants of GPC1 gene reduce risk of biliary atresia in a Chinese population[J]. J Pediatr. Surg, 2016,51(10):1661-1664.DOI:10. 1016/j. jpedsurg. 2016. 05. 009.
13 Melo SA,Luecke LB,Kahlert C,et al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer[J].Nature, 2015,523(7559):177-182. DOI:10. 1038/nature14581.
14 Lu H,Niu F,Liu F,et al. Elevated glypican-1 expression is associated with an unfavorable prognosis in pancreatic ductal adenocarcinoma[J]. Cancer Med, 2017, 6(6):1181-1191. DOI:10. 1002/cam4. 1064.
15 Hara H,Takahashi T,Serada S,et al. Overexpression of glypican4 implicates poor prognosis and their chemoresistance in oesophageal squamous cell carcinoma[J]. Br J Cancer,2016, 115(1):66-75. DOI:10. 1038/bjc. 2016.183.
16 Lai X,Wang M,McElyea SD,et al. A microRNA signature in circulating exosomes is superior to exosomal glypican-4levels for diagnosing pancreatic cancer[J]. Cancer letters,2017,393(1):86-93. DOI:10. 1016/j. canlet. 2017.02.019.
17 Li J,Li B,Ren C,et al. The clinical significance of circulating GPC1 positive exosomes and its regulative miRNAs in colon cancer patients[J]. Oncotarget,2017,8(60):101189-101202. DOI:10.18632/oncotarget. 20516.
18 Chamorro-Jorganes A,Araldi E,Rotllan N,et al. Autoregulation of glypican-1 by intronic microRNA-149 fine tunes the angiogenic response to FGF2 in humanendothelial cells[J]. J Cell Sci,2014,127(6):1169-1178. DOI:10. 1242/jcs. 130518.
2 Wen J, Xiao Y, Wang J, et al. Low doses of CMV induce autoimmune-mediated and inflammatory responses in bile duct epithelia of regulatory T cell-depleted neonatal mice[J]. Lab Invest,2015, 95(2):180-192. DOI:10. 1038/labinvest.2014.148.
3郭振亚,张若岩,柳明江,等.先天性胆道闭锁的病因研究进展[J].临床肝胆病杂志,2015,31(8):1343-1346.DOI:10. 3969/j. issn. 1001-5256.2015.08. 043.Guo ZY, Zhang RY, Liu MG, et al. Etiology of congenital biliary atresia[J]. Journal of Clinical Hepatobiliary Diseases,2015,31(8):1343-1346. DOI:10. 3969/j. issn. 1001-5256.2015.08.043.
4 Tang V,Cofer ZC,Cui S, et al. Loss of a candidate biliary atresia susceptibility gene,add3 a,causes biliary developmental defects in zebrafish[J]. J Pediatr Gastroenterol Nutr,2016,63(5):524-530. DOI:10. 1097/MPG. 0000000000001375.
5李智涵,王斌.Ⅲ型胆道闭锁手术治疗的研究进展[J].临床小儿外科杂志,2016,15(1):11-13. DOI:10.3969/j.issn. 1671-6353.2016.01.004. Li ZH,Wang B. Advances in surgical treatment of biliary atresia of typeⅢ[J]. J Clin Ped Sur,2016,15(1):11-13. DOI:10. 3969/j. issn. 1671-6353.2016.01.004.
6 Asai A, Miethke A, Bezerra JA. Pathogenesis of biliary atresia:defining biology to understand clinical phenotypes[J].Nat Rev Gastroenterol Hepatol,2015,12(6):342-352. DOI:10. 1038/nrgastro. 2015. 74.
7 Hill JJ, Tremblay TL,Fauteux F,et al. Glycoproteomic comparison of clinical triple-negative and luminal breast tumors[J]. Journal of Proteome Research,2015,14(3):1376-1388. DOI:10. 1021/pr500987r.
8 Sadek KH,Ezzat S, Abdel-Aziz SA,et al. Macrophage migration inhibitory factor(MIF)gene promotor polymorphism is associated with increased fibrosis in biliary atresia patients,but not with disease susceptibility[J]. Ann Hum Genet,2017,81(5):177-183.DOI:10. 1111/ahg. 12199.
9 Tang V,Cofer ZC,Cui S,et al. Loss of a candidate biliary atresia susceptibility gene, add3 a, causes biliary developmental defects in zebrafish[J]. J Pediatr Gastroenterol Nutr,2016,63(5):524-530. DOI:10. 1097/MPG. 0000000000001375.
10 Leyva-Vega M, Gerfen J, Thiel BD,et al. Genomic alterations in biliary atresia suggest region of potential disease susceptibility in 2q37. 3[J]. Am J. Med. Genet. A, 2010,152A(4):886-895. DOI:10.1002/ajmg. a. 33332.
11 Cui S,Leyva-Vega M,Tsai EA,et al. Evidence from human and zebrafish that GPC1 is a biliary atresia susceptibility gene[J]. Gastroenterology,2013,144(5):1107-1115.DOI:10. 1053/j. gastro. 2013.01.022.
12 Ke J,Zeng S,Mao J,et al. Common genetic variants of GPC1 gene reduce risk of biliary atresia in a Chinese population[J]. J Pediatr. Surg, 2016,51(10):1661-1664.DOI:10. 1016/j. jpedsurg. 2016. 05. 009.
13 Melo SA,Luecke LB,Kahlert C,et al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer[J].Nature, 2015,523(7559):177-182. DOI:10. 1038/nature14581.
14 Lu H,Niu F,Liu F,et al. Elevated glypican-1 expression is associated with an unfavorable prognosis in pancreatic ductal adenocarcinoma[J]. Cancer Med, 2017, 6(6):1181-1191. DOI:10. 1002/cam4. 1064.
15 Hara H,Takahashi T,Serada S,et al. Overexpression of glypican4 implicates poor prognosis and their chemoresistance in oesophageal squamous cell carcinoma[J]. Br J Cancer,2016, 115(1):66-75. DOI:10. 1038/bjc. 2016.183.
16 Lai X,Wang M,McElyea SD,et al. A microRNA signature in circulating exosomes is superior to exosomal glypican-4levels for diagnosing pancreatic cancer[J]. Cancer letters,2017,393(1):86-93. DOI:10. 1016/j. canlet. 2017.02.019.
17 Li J,Li B,Ren C,et al. The clinical significance of circulating GPC1 positive exosomes and its regulative miRNAs in colon cancer patients[J]. Oncotarget,2017,8(60):101189-101202. DOI:10.18632/oncotarget. 20516.
18 Chamorro-Jorganes A,Araldi E,Rotllan N,et al. Autoregulation of glypican-1 by intronic microRNA-149 fine tunes the angiogenic response to FGF2 in humanendothelial cells[J]. J Cell Sci,2014,127(6):1169-1178. DOI:10. 1242/jcs. 130518.