Cancer stem cells in liver metastasis from colon adenocarcinoma express components of the renin-angiotensin system

Abstract

Aim: We have recently identified a cancer stem cell (CSC) subpopulations within the tumor nests (TNs) and another within the peritumoral stroma (PTS) in liver metastasis from colon adenocarcinoma (LMCA). This study investigated the expression of components of the renin-angiotensin (RAS): pro-renin receptor (PRR), angiotensin converting enzyme (ACE), and angiotensin II receptor 1 (ATIIR1) and angiotensin II receptor 2 (ATIIR2) in LMCA.

Methods: 3,3-Diaminobenzidine (DAB) immunohistochemical (IHC) staining was performed on 16 LMCA samples for PRR, ACE, ATIIR1 and ATIIR2. Immunofluorescence (IF) IHC staining was performed to investigate co-expression of these components of the RAS with SOX2 or OCT4. NanoString analysis (n = 6) and Western blotting (WB, n = 3) were performed on snap-frozen LMCA samples to confirm mRNA and protein expression, respectively.

Results: DAB IHC staining showed the expression of PRR, ACE, ATIIR1 and ATIIR2 within all LMCA samples. NanoString analysis and WB confirmed gene and protein expression of these components of the RAS. IF IHC staining demonstrated expression of PRR, ATIIR1 and ATIIR2 by the SOX2⁺ CSCs within the TNs and the OCT4⁺ CSCs within the PTS. ACE was expressed on the endothelium of the microvessels within the PTS.

Conclusion: These finding suggests the CSCs within LMCA maybe a novel therapeutic target by manipulation of the RAS.

Keywords

Cancer stem cells / colorectal / colon / cancer / adenocarcinoma / liver / metastasis / renin-angiotensin system

References

[1] Aran V,Thuler LC.Colorectal cancer: epidemiology, disease mechanisms and interventions to reduce onset and mortality..Clin Colorectal Cancer2016;15:195-203

[2] New Zealand Ministry of Health2016;WellingtonMinistry of Health

[3] Garden OJ,Poston GJ,Saunders M.Guidelines for resection of colorectal cancer liver metastases..Gut2006;55:iii1-8 PMCID:PMC1860000

[4] Fearon ER.Molecular genetics of colorectal cancer..Annu Rev Pathol2011;6:479-507

[5] Haggar FA.Colorectal cancer epidemiology: incidence, mortality, survival, and risk factors..Clin Colon and Rectal Surg2009;22:191-7 PMCID:PMC2796096

[6] Weiss L,Torhorst J,Moberg I.Haematogenous metastatic patterns in colonic carcinoma: an analysis of 1541 necropsies..J Pathol1986;150:195-203

[7] Scheele J,Altendorf-Hofmann A.Resection of colorectal liver metastases..World J Surg1995;19:59-71

[8] Grillet F,Villeronce O,Lagerqvist EL.Circulating tumour cells from patients with colorectal cancer have cancer stem cell hallmarks in ex vivo culture..Gut2017;66:1802-10 PMCID:PMC5595103

[9] Lahr CJ,Cloud G,Urist MM.A multifactorial analysis of prognostic factors in patients with liver metastases from colorectal carcinoma..J Clin Oncol1983;1:720-6

[10] Stangl R,Charnley RM.Factors influencing the natural history of colorectal liver metastases..Lancet1994;343:1405-10

[11] Cady B.The role of surgical resection of liver metastases in colorectal carcinoma..Semin Oncol1991;18:399-406

[12] Zalinski S,Farges O.Management of patients with synchronous liver metastases of colorectal cancer. Clinical practice guidelines. Guidelines of the French society of gastrointestinal surgery (SFCD) and of the association of hepatobiliary surgery and liver transplantation (ACHBT). Short version..J Visc Surg2011;148:e171-82

[13] Shaheen Mousa A,Nada OH.Immunohistochemical expression of stem cell markers CD133 and Oct4 in colorectal adenocarcinoma..Egyptian Journal of Pathology2014;34:44-51

[14] Baillie R,Yu HH,Davis PF.Cancer stem cells in moderately differentiated oral tongue squamous cell carcinoma..J of Clin Pathol2016;69:742-4 PMCID:PMC4975854

[15] Yu HH,Tan ST,Brasch HD.Characterization of cancer stem cells in moderately differentiated buccal mucosal squamous cell carcinoma..Front Surg2016;3:46 PMCID:PMC4970507

[16] Ram RS,Dunne JC,Tan ST.Cancer stem cells in moderately differentiated lip squamous cell carcinoma express components of the renin-angiotensin system..Front Surg2017;4:30 PMCID:PMC5459876

[17] Bradshaw AR,Brasch HD,Davis PF.Glioblastoma multiforme cancer stem cells express components of the renin-angiotensin system..Front Surg2016;3:51 PMCID:PMC5037176

[18] Noto Z,Okabe M,Fathy M.CD44 and SSEA-4 positive cells in an oral cancer cell line HSC-4 possess cancer stem-like cell characteristics..Oral Oncology49:787-95

[19] Elsir T,Carlson J,Bergqvist M.A study of embryonic stem cell-related proteins in human astrocytomas: identification of nanog as a predictor of survival..Int J Cancer2014;134:1123-31

[20] Chinn S,Peters R.The role of head and neck squamous cell carcinoma cancer stem cells in tumorigenesis, metastasis, and treatment failure..Front Endocrinol (Lausanne)2012;3:90 PMCID:PMC3411100

[21] Humphries H,Marsh RW,Mehrotra S.Characterisation of cancer stem cells in metastatic colon adenocarcinoma to the liver..Front Surg2018;4:76 PMCID:PMC5786574

[22] Munro MJ,Davis PF,Tan ST.Renin-angiotensin system and cancer: a review..Integr Cancer Sci Ther2017;

[23] Nguyen G.Renin, (pro) renin and receptor: an update..Clin Sci (Lond)2011;120:169-78

[24] Ager EI,Christophi C.The renin-angiotensin system and malignancy..Carcinogenesis2008;29:1675-84

[25] Childers WK.Interactions of the renin-angiotensin system in colorectal cancer and metastasis..Int J Colorectal Dis2015;30:749-52

[26] Heinzerling JH,Livingston EH.Predictors of distant metastasis and mortality in patients with stage II colorectal cancer..Am Surg2007;73:230-8

[27] Featherston T,Dunne JC,Brasch HD.Cancer stem cells in moderately differentiated buccal mucosal squamous cell carcinoma express components of the renin–angiotensin system..Front Surg2016;3:52 PMCID:PMC5037224

[28] Itinteang T,Chibnall AM,Davis PF.Cancer stem cells in moderately differentiated oral tongue squamous cell carcinoma express components of the renin-angiotensin system..J Clin Pathol2016;69:942-5 PMCID:PMC5050289

[29] Sun H,Zhuang R,Zheng Y.Do renin-angiotensin system inhibitors influence the recurrence, metastasis, and survival in cancer patients? Evidence from a meta-analysis including 55 studies..Medicine (Baltimore)2017;96:e6394 PMCID:PMC5380250

[30] Tan EMS,Chudakova DA,Marsh R.Characterisation of lymphocyte subpopulations in infantile haemangioma..J Clin Pathol2015;68:812-8

[31] Tan K,van Schaijik B,Marsh RW.Expression and Localization of Cathepsins B, D, and G in Dupuytren’s Disease..Plast Reconstr Surg Glob Open 90002018;6:e1686 PMCID:PMC5865920

[32] Harada K,Fujimoto N,Hasegawa Y.Localization of type-2 angiotensin ii receptor in adrenal gland..J Histochem Cytochem2010;58:585-93 PMCID:PMC2889401

[33] Hallersund P,Helander HF.The expression of renin-angiotensin system components in the human gastric mucosa..J Renin Angiotensin Aldosterone Syst2011;12:54-64

[34] Mehrotra S,Schaijik BV,Marsh RW.Expression and localization of cathepsins B, D and G in cancer stem cells in liver metastasis from colon adenocarcinoma..Front Surg2018;5:40 PMCID:PMC6110174

[35] Wen SW,Neo J.The renin angiotensin system regulates Kupffer cells in colorectal liver metastases..Cancer Biol Ther2013;14:720-7 PMCID:PMC3841211

[36] Neal B,Chapman N.Effects of ACE inhibitors, calcium antagonists, and other blood-pressure-lowering drugs: results of prospectively designed overviews of randomised trials..Lancet2000;356:1955-64

[37] Oh BH,Herron JR,Khan M.Aliskiren, an oral renin inhibitor, provides dose-dependent efficacy and sustained 24-hour blood pressure control in patients with hypertension..J Am Coll Cardiol2007;49:1157-63

[38] Burnier M.Angiotensin II receptor antagonists..Lancet2000;355:637-45

[39] Koh SP,Brasch HD,Tan ST.Expression of cathepsins B, D, and G in isocitrate dehydrogenase-wildtype glioblastoma..Front Surg2017;4:28 PMCID:PMC5447023