Evaluation of CD44 and CD133 as markers of liver cancer stem cells in Egyptian patients with HCV-induced chronic liver diseases versus hepatocellular carcinoma
Keywords:
hepatocellular carcinoma, Cancer stem cell, CD133, CD44, METAVIRAbstract
Background: Cancer stem cells (CSCs) play a critical role in tumor development, progression, metastasis and recurrence.
Aim: To evaluate hepatic expression of CD44 and CD133 in Egyptian patients with HCV-induced chronic liver diseases and hepatocellular carcinomas (HCCs), and to assess its correlation with inflammatory activity scores, stages of fibrosis (in chronic hepatitis with or without cirrhosis) and grades of HCC.
Methods: This prospective case-control study was conducted on eighty subjects who attended the Tropical Diseases Department, Al-Azhar University Hospital, and in collaboration with Theodor Bilharz Research Institute (2014-2016). They were divided as follows: A) Control healthy group: Ten individuals with serologically negative HCV-Ab and HBsAg, and histopathologically normal liver, B) Seventy patients subdivided into 3 groups; Twenty subjects each, as: HCV-Ab+ non-cirrhotic, HCV-Ab+ cirrhotic and HCC. Necroinflammatory activity and fibrosis in non-neoplastic liver biopsies were scored according to the METAVIR scoring system. CD44 and CD133 immunostaining was evaluated in all groups semi-quantitatively using H score. Statistical analysis was performed by SPSS version 22, using independent-samples t-test.
Results: Our study showed a significant increase of mean CD44 & CD133 expression values with disease progression among the groups (p<0.05). Their expressions increased significantly with the inflammatory activity scores and stages of fibrosis, reaching the highest values in A3F4 score compared to A1F1 (p<0.05). Moreover, there was a significant increase of their expressions across HCC grades (p<0.05), however with no significant correlation with focal lesions size.
Conclusion: CSCs clusters exhibiting CD133+ and/or CD44+ profiles were identified in chronic hepatitis, liver cirrhosis and HCC. CD133 and CD44 expressions significantly corresponded to the increased inflammatory activity, fibrosis stages and higher tumor grades. Therefore, evaluation of CD44 and CD133 expression profiles as CSCs markers in non-neoplastic liver and HCCs can help in development of novel therapeutic agents for HCC targeting and prevention.
References
Dai L, Ren P, Liu M, Imai H, Tan EM, Zhang JY. Using immunomic approach to enhance tumor- associated autoantibody detection in diagnosis of hepatocellular carcinoma. Clin Immunol. 2014; 152(1-2):
–39. doi: 10.1016/j.clim.2014.03.007. PMID: 24667685, PMCID: PMC4096568.
American Cancer Society. Cancer Facts and Figures. Atlanta, Ga: American Cancer Society; 2014.
El-Serag HB, Rudolph KL. Hepatocellular carcinoma: epidemiology and molecular carcinogenesis.
Gastroenterology. 2007; 132(7): 2557-76. doi: 10.1053/j.gastro.2007.04.061. PMID: 17570226.
Ismail AM, Ziada HN, Sheashaa HA, Shehab El-Din. AB Decline of viral hepatitis prevalence among
asymptomatic Egyptian blood donors: a glimmer of hope. Eur J Intern Med. 2009; 20(5): 490-3. doi:
1016/j.ejim.2009.03.005. PMID: 19712851.
Gomaa AI, Hashim MS, Waked I. Comparing staging systems for predicting prognosis and survival in
patients with hepatocellular carcinoma in Egypt. PLoS One. 2014; 9(3): e90929. doi:
1371/journal.pone.0090929. PMID: 24603710, PMCID: PMC3946382.
El-Serag HB. Epidemiology of hepatocellular carcinoma. Clin Liver Dis. 2001; 5(1): 87-107. doi:
1016/S1089-3261(05)70155-0.
Bruix J, Colombo M. Hepatocellular carcinoma: current state of the art in diagnosis and treatment. Best
Pract Res Clin Gastroenterol. 2014; 28: 751. doi: 10.1016/j.bpg.2014.08.010. PMID: 25260305.
Nishi M, Sakai Y, Akutsu H, Nagashima Y, Quinn G, Masui S, et al. Induction of cells with cancer stem
cell properties from nontumorigenic human mammary epithelial cells by defined reprogramming factors.
Oncogene. 2014; 33(5): 643-52. doi: 10.1038/onc.2012.614. PMID: 23318426, PMCID: PMC4697746.
Xu XL, Xing BC, Han HB, Zhao W, Hu MH, Xu ZL, et al. The properties of tumor-initiating cells from a
hepatocellular carcinoma patient's primary and recurrent tumor. Carcinogenesis. 2010; 31(2): 167-74. doi:
1093/carcin/bgp232. PMID: 19897602.
Sun JH, Luo Q, Liu LL, Song GB. Liver cancer stem cell markers: Progression and therapeutic
implications. World J Gastroenterol. 2016; 22(13): 3547-57. doi: 10.3748/wjg.v22.i13.3547. PMID:
, PMCID: PMC4814640.
Jaggupilli A, Elkord E. Significance of CD44 and CD24 as Cancer Stem Cell Markers: An Enduring
Ambiguity. Clinical and Developmental Immunology. 2012; 2012. doi: 10.1155/2012/708036.
Naor D, Nedvetzki S, Golan I, Melnik L, Faitelson Y. CD44 in cancer. Crit Rev Clin Lab Sci. 2002; 39(6):
-57. doi: 10.1080/10408360290795574. PMID: 12484499.
Al-Hajj M, Wicha MS, Benito-Hernandez A, Morrison SJ, Clarke MF. Prospective identification of
tumorigenic breast cancer cells. Proc Natl Acad Sci U S A. 2003; 100(7): 3983-8. doi:
1073/pnas.0530291100. PMID: 12629218, PMCID: PMC153034.
Collins AT, Berry PA, Hyde C, Stower MJ, Maitland NJ. Prospective identification of tumorigenic prostate
cancer stem cells. Cancer Res. 2005; 65(23): 10946-51. doi: 10.1158/0008-5472.CAN-05-2018. PMID: 16322242.
Dalerba P, Dylla SJ, Park IK, Liu R, Wang X, Cho RW, et al. Phenotypic characterization of human
colorectal cancer stem cells. Proc Natl Acad Sci U S A. 2007; 104(24): 10158-63. doi:
1073/pnas.0703478104. PMID: 17548814, PMCID: PMC1891215.
Dan T, Hewitt SM, Ohri N, Ly D, Soule BP, Smith SL, et al. CD44 is prognostic for overall survival in the
NCI randomized trial on breast conservation with 25 year follow-up. Breast Cancer Res Treat. 2014;
(1): 11-8. doi: 10.1007/s10549-013-2758-9. PMID: 24276281.
Galizia G, Gemei M, Del Vecchio L, Zamboli A, Di Noto R, Mirabelli P, et al. Combined CD133/CD44
expression as a prognostic indicator of disease-free survival in patients with colorectal cancer. Arch Surg.
; 147(1): 18-24. doi: 10.1001/archsurg.2011.795. PMID: 22250106.
Tovuu LO, Imura S, Utsunomiya T, Morine Y, Ikemoto T, Arakawa Y, et al. Role of CD44 expression in
non-tumor tissue on intrahepatic recurrence of hepatocellular carcinoma. Int J Clin Oncol. 2013; 18(4):
-6. doi: 10.1007/s10147-012-0432-6. PMID: 22706704.
Sun J, Luo Q, Liu L, Zhang B, Shi Y, Ju Y, et al. Biomechanical profile of cancer stem-like cells derived
from MHCC97H cell lines. J Biomech. 2016; 49(1): 45-52. doi: 10.1016/j.jbiomech.2015.11.007. PMID:
Grosse-Gehling P, Fargeas CA, Dittfeld C, Garbe Y, Alison MR, Corbeil D, et al. CD133 as a biomarker
for putative cancer stem cells in solid tumours: limitations, problems and challenges. J Pathol. 2013;
(3): 355-78. doi: 10.1002/path.4086. PMID: 22899341.
Vander Griend DJ, Karthaus WL, Dalrymple S, Meeker A, DeMarzo AM, Isaacs JT. The role of CD133 in
normal human prostate stem cells and malignant cancer-initiating cells. Cancer Res. 2008; 68(23): 9703-11.
doi: 10.1158/0008-5472.CAN-08-3084. PMID: 19047148, PMCID: PMC3072758.
Shmelkov SV, Butler JM, Hooper AT, Hormigo A, Kushner J, Milde T, et al. CD133 expression is not
restricted to stem cells, and both CD133+ and CD133-metastatic colon cancer cells initiate tumors. J Clin
Invest. 2008; 118(6): 2111-20. doi: 10.1172/JCI34401. PMID: 18497886, PMCID: PMC2391278.
Qin Q, Sun Y, Fei M, Zhang J, Jia Y, Gu M, et al. Expression of putative stem marker nestin and CD133 in
advanced serous ovarian cancer. Neoplasma. 2012; 59(3): 310-5. doi: 10.4149/neo_2012_040. PMID:
Suetsugu A, Nagaki M, Aoki H, Motohashi T, Kunisada T, Moriwaki H. Characterization of CD133+
hepatocellular carcinoma cells as cancer stem/progenitor cells. Biochem Biophys Res Commun. 2006;
(4): 820-4. doi: 10.1016/j.bbrc.2006.10.128. PMID: 17097610.
Liu H, Zhang W, Jia Y, Yu Q, Grau GE, Peng L, et al. Single-cell clones of liver cancer stem cells have the
potential of differentiating into different types of tumor cells. Cell Death Dis; 2013; 4: e857. doi:
1038/cddis.2013.340. PMID: 24136221, PMCID: PMC3824650.
Bedossa P, Poynard T. An Algorithm for the Grading of Activity in Chronic Hepatitis C. The METAVIR
Cooperative Study Group. Hepatology. 1996; 24(2): 289–93. doi: 10.1002/hep.510240201. PMID:
Hsu SM, Raine L. Protein A, avidin, and biotin in immunohistochemistry. J Histochem Cytochem. 1981;
(11): 1349-53. doi: 10.1177/29.11.6172466. PMID: 6172466.
Itoi H, Fujimori Y, Tsutsui H, Matsui K, Hada T, Kakishita E, et al. Differential upregulation of
interleukin-18 receptor alpha chain between CD4+ and CD8+ T cells during acute graft-versus-host disease
in mice. J Interferon Cytokine Res. 2004; 24(5): 291-6. doi: 10.1089/107999004323065075. PMID:
Machlin ES, Sarnow P, Sagan SM. Masking the 5' terminal nucleotides of the hepatitis C virus genome by
an unconventional microRNA-target RNA complex. Proc Natl Acad Sci U S A. 2011; 108(8): 3193-8. doi:
1073/pnas.1012464108. PMID: 21220300, PMCID: PMC3044371.
Nguyen MH, Keeffe EB. Prevalence and treatment of hepatitis C virus genotypes 4, 5, and 6. Clin
Gastroenterol Hepatol. 2005; 3(10 Suppl 2): S97-S101. doi: 10.1016/S1542-3565(05)00711-1. PMID:
Bartenschlager R, Penin F, Lohmann V, André P. Assembly of infectious hepatitis C virus particles. Trends
Microbiol. 2011; 19: 95–103. doi: 10.1016/j.tim.2010.11.005. PMID: 21146993.
Siegel R, Ma J, Zou Z, Jemal A. Cancer statistics, 2014. CA Cancer J Clin. 2014; 64(1): 9-29. doi:
3322/caac.21208. PMID: 24399786.
Malanchi I, Santamaria-Martínez A, Susanto E, Peng H, Lehr HA, Delaloye JF, et al. Interactions between
cancer stem cells and their niche govern metastatic colonization. Nature. 2011; 481(7379): 85–9. doi:
1038/nature10694. PMID: 22158103.
Eyler CE, Rich JN. Survival of the fittest: cancer stem cells in therapeutic resistance and angiogenesis. J
Clin Oncol. 2008; 26(17): 2839–45. doi: 10.1200/JCO.2007.15.1829. PMID: 18539962, PMCID: PMC2739000. 35) Zhao Q, Zhou H, Liu Q, Cao Y, Wang G, Hu A, et al. Prognostic value of the expression of cancer stem
cell-related markers CD133 and CD44 in hepatocellular carcinoma: From patients to patient-derived tumor
xenograft models. Oncotarget. 2016; 7(30): 47431-43. doi: 10.18632/oncotarget.10164. PMID: 27329727,
PMCID: PMC5216952.
Zhu Z, Hao X, Yan M, Yao M, Ge C, Gu J, et al. Cancer stem/progenitor cells are highly enriched in
CD133 (+) CD44 (+) population in hepatocellular carcinoma. Int J Cancer. 2010; 126(9): 2067-78. doi:
1002/ijc.24868. PMID: 19711346.
Ma S, Chan KW, Hu L, Lee TK, Wo JY, Ng IO, et al. Identification and characterization of tumorigenic
liver cancer stem/progenitor cells. Gastroenterology. 2007; 132(7): 2542–56. doi:
1053/j.gastro.2007.04.025. PMID: 17570225.
Mansour AH, Elkhodary TR, Anwar R, Habeeb MR, Mohammed MA. Regulation of cancer stem cell
marker (CD133) by Transforming Growth Factor Beta in hepatocellular carcinoma. International Journal of
Cancer research. 2014; 10(2): 65-73. doi: 10.3923/ijcr.2014.65.73.
Endo K, Terada T. Protein expression of CD44 (standard and variant isoforms) in hepatocellular
carcinoma: relationships with tumor grade, clinicopathologic parameters, p53 expression, and patient
survival. J Hepatol. 2000; 32(1): 78–84. doi: 10.1016/S0168-8278(00)80192-0. PMID: 10673070.
Luo Y, Tan Y. Prognostic value of CD44 expression in patients with hepatocellular carcinoma:
meta‑analysis. Cancer Cell Int. 2016; 16: 47. doi: 10.1186/s12935-016-0325-2. PMID: 27330410
Yoshikawa S, Zen Y, Fujii T, Sato Y, Ohta T, Aoyagi Y, et al. Characterization of CD133+ parenchymal
cells in the liver: histology and culture. World J Gastroenterol. 2009; 15(39): 4896-906. doi:
3748/wjg.15.4896. PMID: 19842219, PMCID: PMC2764966.
Zeng Z, Ren J, O'Neil M, Zhao J, Bridges B, Cox J, et al. Impact of stem cell marker expression on
recurrence of TACE-treated hepatocellular carcinoma post liver transplantation. BMC Cancer. 2012; 12:
doi: 10.1186/1471-2407-12-584. PMID: 23216644, PMCID: PMC3534240.
Sabet MN, Rakhshan A, Erfani E, Madjd Z. Co-Expression of Putative Cancer Stem Cell Markers, CD133
and Nestin, in Skin Tumors. Asian Pac J Cancer Prev. 2014; 15: 8161–9. PMID: 25339000.
Gandara DR, Lara PN Jr, Mack PC. Patient-Derived Xenografts for Investigation of Acquired Resistance in
Oncogene-Driven Cancers: Building a Better Mousetrap. J Clin Oncol. 2015; 33(26): 2839-40. doi:
1200/JCO.2015.61.9692. PMID: 26215942.
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