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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front. Med.    2019, Vol. 13 Issue (4) : 482-491
G protein-coupled receptor LGR6 is an independent risk factor for colon adenocarcinoma
Wenjing Wang1, Shigang Ding1(), Hejun Zhang1, Jun Li1, Jun Zhan2(), Hongquan Zhang2()
1. Department of Gastroenterology, Peking University Third Hospital, Beijing 100191, China
2. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), and State Key Laboratory of Natural and Biomimetic Drugs, and Department of Anatomy, Histology and Embryology, Peking University Health Science Center, Beijing 100191, China
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LGR6 is a member of the G protein-coupled receptor family that plays a tumor-suppressive role in colon cancer. However, the relationship between LGR6 expression in patients and clinicopathological factors remains unclear. This study aimed to clarify whether the expression level of LGR6 is correlated with colon adenocarcinoma progression. Immunohistochemistry was used to detect LGR6 expression in colon adenoma tissues (n = 21), colon adenocarcinoma tissues (n = 156), and adjacent normal tissues (n = 124). The expression levels of LGR6 in colon adenoma and adenocarcinoma were significantly higher than those in normal colon epithelial tissues (P<0.001). Low LGR6 expression predicted a short overall survival in patients with colon adenocarcinoma (log-rank test, P = 0.016). Univariate and multivariate survival analyses showed that, in addition to N and M classification, LGR6 expression served as an independent prognostic factor. Thus, low expression of LGR6 can be used as an independent prognostic parameter in patients with colon adenocarcinoma.

Keywords LGR6      colon adenocarcinoma      immunohistochemistry      prognosis     
Corresponding Authors: Shigang Ding,Jun Zhan,Hongquan Zhang   
Just Accepted Date: 28 April 2018   Online First Date: 02 July 2018    Issue Date: 02 August 2019
 Cite this article:   
Wenjing Wang,Shigang Ding,Hejun Zhang, et al. G protein-coupled receptor LGR6 is an independent risk factor for colon adenocarcinoma[J]. Front. Med., 2019, 13(4): 482-491.
Characteristic Value/number of patients Ratio
Age median (range), year 67 (24–91)
Male 85 54.50%
Female 71 45.50%
Age, year
<60 30 19.20%
60–91 126 80.80%
Tumor Nodes Metastases category
T1 4 2.60%
T2 9 5.80%
T3 113 72.40%
T4 30 19.20%
N0 85 54.50%
N1 54 34.60%
N2 17 10.90%
M0 149 95.50%
M1 7 4.50%
American Joint Committee on Cancer category
I 11 7.10%
II 73 46.80%
III 65 41.70%
IV 7 4.50%
Survival status
Alive 88 56.40%
Dead 68 43.60%
Tab.1  Clinicopathological factors of patients with colon adenocarcinoma (n = 156)
Fig.1  LGR6 is expressed in colon adenocarcinoma. Immunohistochemical staining was performed to examine LGR6 expression in colon adenocarcinomas. Representative photographs are shown, and corresponding regions are enlarged. Staining scores were categorized into four grades: (A) no staining (0); (B) weak staining (1+); (C) moderate staining (2+); (D) strong staining (3+).
Fig.2  Expression of LGR6 in normal, adenoma, and adenocarcinoma colon tissues. (A) Normal colon mucosa; (B) large tissue section enclosing adenoma and normal colon mucosa; (C) normal tissue extracted from Fig. 2B; (D) adenoma tissue extracted from Fig. 2B; (E) colon adenocarcinoma tissue; (F) LGR6 expression is higher in colon adenoma and adenocarcinoma tissues than in normal colon mucosa (t-test, P<0.001), whereas differences between adenoma and adenocarcinoma are insignificant (t-test, P>0.05).
Fig.3  Expression of LGR6 in colorectal adenoma and adenocarcinoma. (A) LGR6 expression in Sabates–Bellver colon data set in Oncomine: rectal and colonic adenomas highly express LGR6; (B) LGR6 expression in Skrzypczak colorectal data set in Oncomine: LGR6 expression in colorectal adenoma is significantly higher than that in normal tissue; (C and D) relationships between KRAS and LGR6 expression in normal and tumor tissues, respectively, in the UCSC database (P = 0.1810; P = 0.2177). (E) LGR6 expression is significantly higher in KRAS mutant than in KRAS wild-type specimens in the Oncomine database (P = 0.0006). (F) LGR6 expression is lower in specimens with microsatellite than in those with microsatellite stability in Oncomine (P = 0.0220).
Characteristic Low-expression group High-expression group P
0 (0–1) (1–2) (2–3)
No. of patients Ratio No. of patients Ratio No. of patients Ratio No. of patients Ratio
Gender 0.271
Male 5 3.20% 31 19.90% 31 19.90% 18 11.50%
Female 7 4.50% 16 10.30% 30 19.20% 18 11.50%
Age, year 0.420
<60 2 1.30% 11 7.10% 8 5.10% 9 5.80%
60–91 10 6.40% 36 23.10% 53 34.00% 27 17.30%
Tumor Nodes Metastases category
T1 1 0.60% 3 1.90% 0 0.00% 0 0.00% 0.068a
T2 1 0.60% 1 0.60% 6 3.80% 1 0.60% 0.123b
T3 8 5.10% 32 20.50% 42 26.90% 31 19.90%
T4 2 1.30% 11 7.10% 13 8.30% 4 2.60%
N0 6 3.80% 28 17.90% 26 16.70% 25 16.00% 0.063
N1–N2 6 3.80% 19 12.20% 35 22.40% 11 7.10%
M0 11 7.10% 44 28.20% 58 37.20% 36 23.10% 0.367
M1 1 0.60% 3 1.90% 3 1.90% 0 0.00%
American Joint Committee on Cancer
I 1 0.60% 4 2.60% 5 3.20% 1 0.60% 0.285a
II 5 3.20% 23 14.70% 21 13.50% 24 15.40% 0.146b
III 5 3.20% 17 10.90% 32 20.50% 11 7.10%
IV 1 0.60% 3 1.90% 3 1.90% 0 0.00%
Survival status
Alive 4 2.60% 23 14.70% 36 23.10% 25 16.00% 0.036*
Dead 8 5.10% 24 15.40% 25 16.00% 11 7.10%
Tab.2  Association between LGR6 expression levels and clinical variables in colon adenocarcinoma
Characteristic Univariate survival analysis Multivariate survival analysis
P HR 95% CI P HR 95% CI
Male 0.653 0.896 0.554–1.499
Age, year
<60 0.612 1.174 0.629–2.191
Tumor Nodes Metastases category
T1–T2 0.063 3.794 0.929–15.505
N0 0.002* 2.122 1.307–3.444 0.003* 2.109 1.283–3.467
M0 0.000* 5.437 2.305–12.825 0.002* 4.029 1.681–9.657
American Joint Committee on Cancer
I–II 0.001* 2.229 1.370–3.626
Relative level of LGR6
High expression 0.018* 1.780 1.103–2.872 0.008* 1.919 1.182–3.114
Low expression
Tab.3  Univariate and multivariate analysis of overall survival in 156 patients with colon adenocarcinoma
Fig.4  Elevated expression of LGR6 predicts long overall survival for patients with colon adenocarcinoma. Kaplan–Meier analysis of two groups of patients with colon adenocarcinoma with high or low LGR6 expression was determined by log-rank test (P<0.05), which shows that elevated expression of LGR6 correlates with better overall survival for patients with colon adenocarcinoma.
1 W Chen, R Zheng, H Zeng, S Zhang. The incidence and mortality of major cancers in China, 2012. Chin J Cancer 2016; 35(1): 73 pmid: 27484217
2 H Brenner, M Kloor, CP Pox. Colorectal cancer. Lancet 2014; 383(9927): 1490–1502 pmid: 24225001
3 JR Jass. Classification of colorectal cancer based on correlation of clinical, morphological and molecular features. Histopathology 2007; 50(1): 113–130 pmid: 17204026
4 SD Markowitz, MM Bertagnolli. Molecular origins of cancer: molecular basis of colorectal cancer. N Engl J Med 2009; 361(25): 2449–2460 pmid: 20018966
5 FA Sinicrope, DJ Sargent. Molecular pathways: microsatellite instability in colorectal cancer: prognostic, predictive, and therapeutic implications. Clin Cancer Res 2012; 18(6): 1506–1512 pmid: 22302899
6 F De Sousa E Melo, X Wang, M Jansen, E Fessler, A Trinh, LP de Rooij, JH de Jong, OJ de Boer, R van Leersum, MF Bijlsma, H Rodermond, M van der Heijden, CJ van Noesel, JB Tuynman, E Dekker, F Markowetz, JP Medema, L Vermeulen. Poor-prognosis colon cancer is defined by a molecularly distinct subtype and develops from serrated precursor lesions. Nat Med 2013; 19(5): 614–618 pmid: 23584090
7 FJ Carmona, D Azuara, A Berenguer-Llergo, AF Fernández, S Biondo, J de Oca, F Rodriguez-Moranta, R Salazar, A Villanueva, MF Fraga, J Guardiola, G Capellá, M Esteller, V Moreno. DNA methylation biomarkers for noninvasive diagnosis of colorectal cancer. Cancer Prev Res (Phila) 2013; 6(7): 656–665 pmid: 23694962
8 N Vatandoost, J Ghanbari, M Mojaver, A Avan, M Ghayour-Mobarhan, R Nedaeinia, R Salehi. Early detection of colorectal cancer: from conventional methods to novel biomarkers. J Cancer Res Clin Oncol 2016; 142(2): 341–351 pmid: 25687380
9 I Böhme, AG Beck-Sickinger. Illuminating the life of GPCRs. Cell Commun Signal 2009; 7(1): 16 pmid: 19602276
10 SY Hsu, M Kudo, T Chen, K Nakabayashi, A Bhalla, PJ van der Spek, M van Duin, AJ Hsueh. The three subfamilies of leucine-rich repeat-containing G protein-coupled receptors (LGR): identification of LGR6 and LGR7 and the signaling mechanism for LGR7. Mol Endocrinol 2000; 14(8): 1257–1271 pmid: 10935549
11 W de Lau, N Barker, TY Low, BK Koo, VS Li, H Teunissen, P Kujala, A Haegebarth, PJ Peters, M van de Wetering, DE Stange, JE van Es, D Guardavaccaro, RB Schasfoort, Y Mohri, K Nishimori, S Mohammed, AJ Heck, H Clevers. Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling. Nature 2011; 476(7360): 293–297 pmid: 21727895
12 M Leushacke, N Barker. Lgr5 and Lgr6 as markers to study adult stem cell roles in self-renewal and cancer. Oncogene 2012; 31(25): 3009–3022 pmid: 22002312
13 JA Lehoczky, CJ Tabin. Lgr6 marks nail stem cells and is required for digit tip regeneration. Proc Natl Acad Sci USA 2015; 112(43): 13249–13254 pmid: 26460010
14 Y Gao, K Kitagawa, Y Hiramatsu, H Kikuchi, T Isobe, M Shimada, C Uchida, T Hattori, T Oda, K Nakayama, KI Nakayama, T Tanaka, H Konno, M Kitagawa. Up-regulation of GPR48 induced by down-regulation of p27Kip1 enhances carcinoma cell invasiveness and metastasis. Cancer Res 2006; 66(24): 11623–11631 pmid: 17178856
15 JS Steffen, E Simon, V Warneke, K Balschun, M Ebert, C Röcken. LGR4 and LGR6 are differentially expressed and of putative tumor biological significance in gastric carcinoma. Virchows Arch 2012; 461(4): 355–365 pmid: 22855134
16 S Ryuge, Y Sato, SX Jiang, G Wang, M Kobayashi, R Nagashio, K Katono, A Iyoda, Y Satoh, N Masuda. The clinicopathological significance of Lgr5 expression in lung adenocarcinoma. Lung Cancer 2013; 82(1): 143–148 pmid: 23915911
17 E Simon, D Petke, C Böger, HM Behrens, V Warneke, M Ebert, C Röcken. The spatial distribution of LGR5+ cells correlates with gastric cancer progression. PLoS One 2012; 7(4): e35486 pmid: 22530031
18 A Gregorieff, Y Liu, MR Inanlou, Y Khomchuk, JL Wrana. Yap-dependent reprogramming of Lgr5+ stem cells drives intestinal regeneration and cancer. Nature 2015; 526(7575): 715–718 pmid: 26503053
19 H Takahashi, H Ishii, N Nishida, I Takemasa, T Mizushima, M Ikeda, T Yokobori, K Mimori, H Yamamoto, M Sekimoto, Y Doki, M Mori. Significance of Lgr5+ve cancer stem cells in the colon and rectum. Ann Surg Oncol 2011; 18(4): 1166–1174 pmid: 21125339
20 F Walker HH Zhang, A Odorizzi, AW Burgess. LGR5 is a negative regulator of tumourigenicity, antagonizes Wnt signalling and regulates cell adhesion in colorectal cancer cell lines. PLoS One 2011; 6(7): e22733 pmid: 21829496
21 F de Sousa E Melo, S Colak, J Buikhuisen, J Koster, K Cameron, JH de Jong, JB Tuynman, PR Prasetyanti, E Fessler, SP van den Bergh, H Rodermond, E Dekker, CM van der Loos, ST Pals, MJ van de Vijver, R Versteeg, DJ Richel, L Vermeulen, JP Medema. Methylation of cancer-stem-cell-associated Wnt target genes predicts poor prognosis in colorectal cancer patients. Cell Stem Cell 2011; 9(5): 476–485 pmid: 22056143
22 A Guinot, F Oeztuerk-Winder, JJ Ventura. miR-17–92/p38a dysregulation enhances Wnt signaling and selects Lgr6+ cancer stem-like cells during lung adenocarcinoma progression. Cancer Res 2016; 76(13): 4012–4022 pmid: 27197183
23 L Blaas, F Pucci, HA Messal, AB Andersson, E Josue Ruiz, M Gerling, I Douagi, B Spencer-Dene, A Musch, R Mitter, L Bhaw, R Stone, D Bornhorst, AK Sesay, J Jonkers, G Stamp, I Malanchi, R Toftgård, A Behrens. Lgr6 labels a rare population of mammary gland progenitor cells that are able to originate luminal mammary tumours. Nat Cell Biol 2016; 18(12): 1346–1356 pmid: 27798604
24 T Sjöblom, S Jones, LD Wood, DW Parsons, J Lin, TD Barber, D Mandelker, RJ Leary, J Ptak, N Silliman, S Szabo, P Buckhaults, C Farrell, P Meeh, SD Markowitz, J Willis, D Dawson, JK Willson, AF Gazdar, J Hartigan, L Wu, C Liu, G Parmigiani, BH Park, KE Bachman, N Papadopoulos, B Vogelstein, KW Kinzler, VE Velculescu. The consensus coding sequences of human breast and colorectal cancers. Science 2006; 314(5797): 268–274 pmid: 16959974
25 TA Chan, S Glockner, JM Yi, W Chen, L Van Neste, L Cope, JG Herman, V Velculescu, KE Schuebel, N Ahuja, SB Baylin. Convergence of mutation and epigenetic alterations identifies common genes in cancer that predict for poor prognosis. PLoS Med 2008; 5(5): e114 pmid: 18507500
26 X Gong, KS Carmon, Q Lin, A Thomas, J Yi, Q Liu. LGR6 is a high affinity receptor of R-spondins and potentially functions as a tumor suppressor. PLoS One 2012; 7(5): e37137 pmid: 22615920
27 X Wang, Q Sun. TP53 mutations, expression and interaction networks in human cancers. Oncotarget 2017; 8(1): 624–643 pmid: 27880943
28 J Zhan, M Niu, P Wang, X Zhu, S Li, J Song, H He, Y Wang, L Xue, W Fang, H Zhang. Elevated HOXB9 expression promotes differentiation and predicts a favourable outcome in colon adenocarcinoma patients. Br J Cancer 2014; 111(5): 883–893 pmid: 25025961
29 H Clevers, R Nusse. Wnt/b-catenin signaling and disease. Cell 2012; 149(6): 1192–1205 pmid: 22682243
30 KS Carmon, X Gong, Q Lin, A Thomas, Q Liu. R-spondins function as ligands of the orphan receptors LGR4 and LGR5 to regulate Wnt/b-catenin signaling. Proc Natl Acad Sci USA 2011; 108(28): 11452–11457 pmid: 21693646
31 R Lappano, M Maggiolini. G protein-coupled receptors: novel targets for drug discovery in cancer. Nat Rev Drug Discov 2011; 10(1): 47–60 pmid: 21193867
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