Abstract
In the present study, we investigated expression pattern of Cholecystokinin type A receptor (CCKAR) in relation to its commonly studied polymorphism (rs1800857, T/C) in gallstone disease (GSD) patients and controls. A total of 502 subjects (272 GSD and 230 controls) were enrolled, and genotyping was performed by evaluating restriction fragments of PstI digested DNA. For analyzing expression pattern of CCKAR in relation to polymorphism, gallbladder tissue samples from 80 subjects (GSD—55; control—25) were studied. Expression of CCKAR mRNA was evaluated by reverse transcriptase-PCR and confirmed using real-time PCR. Protein expression was evaluated by enzyme-linked immunosorbent assay. We observed significantly (p < 0.0001) lower expression of CCKAR mRNA and protein in GSD tissues as compared with control. Significantly higher frequency of A1/A1 genotype (C/T transition) (p = 0.0005) was observed for GSD as compared with control. Expression of CCKAR protein was found to be significantly lower (p < 0.0001) in A1/A1 genotype as compared with other genotypes for GSD patients. Perhaps, this is the first report providing evidence of alteration in CCKAR expression in relation to its polymorphism elucidating the molecular pathway of the disease. Additional investigations with lager sample size are needed to confirm these findings.
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Baghel K, Kazmi HR, Raj S, Chandra A, Srivastava RN (2014) Elevated expression of maspin mRNA as a predictor of survival in stage II and III gallbladder cancer cases. Asian Pac J Cancer Prev 15:343–347
Chomczynski P (1993) A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques 15:532–537
Ding X, Lu CY, Mei Y, Liu CA, Shi YJ (2005) Correlation between gene expression of CCK-A receptor and emptying dysfunction of the gallbladder in patients with gallstones and diabetes mellitus. Hepatobiliary Pancreat Dis Int 4:295–298
Juvonen T, Niemelä O, Mäkelä J, Kairaluoma MI (1994) Characteristics of symptomatic gallbladder disease in patients with either solitary or multiple cholesterol gallstones. Hepatogastroenterology 41:263–266
Kazmi HR, Chandra A, Baghel K, Singh A, Nigam J, Parmar D, Mahdi AA, Goel SK, Kumar S (2014) Differential expression of cholecystokinin a receptor in gallbladder cancer in the young and elderly suggests two subsets of the same disease? Biomed Res Int 2014:625695. doi:10.1155/2014/625695
Khanna R, Chansuria R, Kumar M, Shukla HS (2006) Histological changes in gall bladder due to stone disease. Indian J Surg 68:201–204
Lu W-T, Zhang X, Zhang M, Gong S-L, Wei J (2004) Association analysis of the Cholecystokinin type A receptor gene in Schizophrenia. Chin Med J 117:627–629
Marschall HU, Einarsson C (2007) Gallstone disease. J Intern Med 261:529–542
Mathur SK, Duhan A, Singh S, Aggarwal M, Aggarwal G, Sen R, Singh S, Garg S (2012) Correlation of gallstone characteristics with mucosal changes in gall bladder. Trop Gastroenterol 33:39–44
Miller LJ, Holicky EL, Ulrich CD, Wieben ED (1995) Abnormal processing of the human cholecystokinin receptor gene in association with gallstones and obesity. Gastroenterology 109:1375–1380
Miyasaka K, Takata Y, Funakoshi A (2002) Association of cholecystokinin A receptor gene polymorphism with cholelithiasis and the molecular mechanisms of this polymorphism. J Gastroenterol 37(Suppl 14):102–106
Nakeeb A, Comuzzie AG, Martin L, Sonnenberg GE, Swartz-Basile D, Kissebah AH, Pitt HA (2002) Gallstones: genetics versus environment. Ann Surg 235:842–849
Nigam J, Chandra A, Kazmi HR, Parmar D, Singh D, Gupta V (2015) Prognostic significance of survivin in resected gallbladder cancer. J Surg Res 194:57–62
Rai R, Tewari M, Kumar M, Singh TB, Shukla HS (2011) Expression profile of cholecystokinin type-A receptor in gallbladder cancer and gallstone disease. Hepatobiliary Pancreat Dis Int 10:408–414
Randi G, Franceschi S, La Vecchia C (2006) Gallbladder cancer worldwide: geographical distribution and risk factors. Int J Cancer 118:1591–1602
Sato N, Miyasaka K, Suzuki S, Kanai S, Ohta M, Kawanami T, Yoshida Y, Takiguchi S, Noda T, Takata Y, Funakoshi A (2003) Lack of cholecystokinin-A receptor enhanced gallstone formation: a study in CCK-A receptor gene knockout mice. Dig Dis Sci 48:1944–1947
Shaffer EA (2006) Gallstone disease: epidemiology of gallbladder stone disease. Best Pract Res Clin Gastroenterol 20:981–996
Shuai J, Zhang S, Han T, Jiang Y, Lei R, Chen S (1999) Correlation between gene expression of CCK-A receptor and gallbladder emptying in gallstone patients. Zhonghua Wai Ke Za Zhi 37:292–294
Shukla VK, Goel S, Trigun SK, Sharma D (2008) Electrophoretic pattern of proteins in carcinoma of the gallbladder. Eur J Cancer Prev 17:9–12
Singh V, Trikha B, Nain C, Singh K, Bose S (2001) Epidemiology of gallstone disease in Chandigarh: a community-based study. J Gastroenterol Hepatol 16:560–563
Srivastava A, Pandey SN, Dixit M, Choudhuri G, Mittal B (2008) Cholecystokinin receptor A gene polymorphism in gallstone disease and gallbladder cancer. J Gastroenterol Hepatol 23:970–975
Stinton LM, Shaffer EA (2012) Epidemiology of gallbladder disease: cholelithiasis and cancer. Gut Liver 6:172–187
Takata Y, Takeda S, Kawanami T, Takiguchi S, Yoshida Y, Miyasaka K, Funakoshi A (2002) Promoter analysis of human cholecystokinin type-A receptor gene. J Gastroenterol 37:815–820
Takimoto T, Terayama H, Waga C, Okayama T, Ikeda K, Fukunishi I, Iwahashi K (2005) Cholecystokinin (CCK) and the CCKA receptor gene polymorphism and smoking behaviour. Psychiatry Res 133:123–128
Tokunaga Y, Cox KL, Coleman R, Concepcion W, Nakazato P, Esquivel CO (1993) Characterization of cholecystokinin receptors on the human gallbladder. Surgery 113:155–162
Upp JR Jr, Nealon WH, Singh P, Fagan CJ, Jonas AS, Greeley GH Jr, Thompson JC (1987) Correlation of cholecystokinin receptors with gallbladder contractility in patients with gallstones. Ann Surg 205:641–648
Völzke H, Baumeister SE, Alte D, Hoffmann W, Schwahn C, Simon P, John U, Lerch MM (2005) Independent risk factors for gallstone formation in a region with high cholelithiasis prevalence. Digestion 71:97–105
Wei J, Hemmings GP (1999) The CCK-A receptor gene possibly associated with auditory hallucinations in schizophrenia. Eur Psychiatry 14:67–70
Xu HL, Hsing AW, Vogtmann E, Chu LW, Cheng JR, Gao J, Tan YT, Wang BS, Shen MC, Gao YT (2013) Variants in CCK and CCKAR genes to susceptibility to biliary tract cancers and stones: a population-based study in Shanghai, China. J Gastroenterol Hepatol 28:1476–1481
Xu GQ, Xu CF, Chen HT, Liu S, Teng XD, Xu GY, Yu CH (2014) Association of caveolin-3 and cholecystokinin A receptor with cholesterol gallstone disease in mice. World J Gastroenterol 20:9513–9518
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The present work was supported by the research grant from Council of Science and Technology, Lucknow, Uttar Pradesh, India (Grant No. CST/SERPD/D-3429), and Indian Council of Medical Research, New Delhi, India (Grant No. 3/2/2/60/2011/NCDIII).
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Kazmi, H.R., Chandra, A., Nigam, J. et al. Polymorphism and Expression Profile of Cholecystokinin Type A Receptor in Relation to Gallstone Disease Susceptibility. Biochem Genet 54, 665–675 (2016). https://doi.org/10.1007/s10528-016-9745-x
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DOI: https://doi.org/10.1007/s10528-016-9745-x