Gene expression profiles from members of two gene families, chosen from the microarray analysis of reisolated CC531 cells.
Abstract
With estimated 700,000 deaths each year, colorectal carcinoma (CRC) continues to be the fourth leading cause of cancer-related deaths worldwide. Fortunately, the mortality of CRC is considered to be most avertable; hence, it is essential to develop new approaches for more accurate and early diagnosis of primary as well as metastatic CRC, including genetic and biomarker tests. In this regard, the intercellular junctions and the insulin-like growth factor (IGF) axis attract increasing attention, since they are involved in several stages of cancer and for their vital role in regulating cell survival and growth; furthermore, constituents of intercellular junctions and of the IGF axis could be used as tumor and/or metastasis markers, which are becoming the focus of increasing research activities. Our experimental results highlight the importance of gene expression changes in the tight junction proteins claudins, and in the IGF-binding proteins IGFBP3 and IGFBP7. They show additionally that claudins and IGFBPs cannot be simply defined in terms of favoring or antagonizing cancer progression but have additional properties and activities, which become apparent only in the context of liver colonization. Furthermore, their intensive modulation during the initial phase of liver colonization may suggest them as early metastasis-related markers.
Keywords
- colorectal cancer
- liver metastasis
- personalized medicine
- influence on treatment
- claudins
- IGFBPs
- tumor cell reisolation
- metastasis marker
1. Introduction
With estimated 700,000 deaths each year, colorectal carcinoma (CRC) continues to be the fourth leading cause of cancer-related deaths in both sexes worldwide [1]. The 5-year relative survival rate for stage IV metastatic CRC is about 11%, while in stage I this number rises to nearly 90%. These figures reflect the fact that despite the high incidence and mortality rate of CRC, its mortality is among the most avertable ones. In addition, the fact that liver metastasis is the cause of most deaths from CRC [2], underlines the significance of (early) metastasis prevention. In other words, it is of great importance to develop new approaches for more accurate and early diagnosis of primary CRC on one hand and of its metastasis on the other; including screening programs as well as genetic, molecular and biomarker tests.
Colorectal cancer progression is driven by increasing or recurring growth of the primary carcinoma as well as hematogenic and lymphatic spread. For hematogenic spread, the liver is most important as it constitutes the first vascular bed in which disseminating CRC cells can be trapped after their dissemination. Hence, this organ is affected in up to 10–20% of CRC patients at the time of presentation. Another 20–25% will develop overt liver metastasis during the course of their illness [3, 4].
The main purpose of our experimental studies was first to develop a suitable model for investigating the efficacy of novel drugs [5, 6, 7]. One of the few well-characterized animal models for hepatic CRC benefits from the rat CC531 cell line. After injecting the cells, liver metastases develop and their growth has been frequently used for studying effects of various anti-cancer treatments [8, 9, 10].
The second aim was to identify genes, which are instrumental in the survival and metastasis formation of disseminated CRC cells. In addition, we reasoned that there are genes, which are necessary for the primary tumor as well as those, which are essential for metastasis initiation and formation. We furthermore hypothesized that the latter genes would be modulated in expression during the cells’ colonization of the liver. Consequently, temporal changes in gene expression of CRC cells homing into the liver were investigated using an
The National Institutes of Health defined a biomarker as “a characteristic that is objectively measured and evaluated as an indicator of normal biological processes, pathogenic processes, or pharmacologic responses to a therapeutic intervention [11]. From a therapeutic point of view, genome variations are recognized as the main cause of variable response to and side effects of drugs and a “one size fits all” approach is not the best solution any more. The individual’s genetic and molecular makeup will be devoted to improve and develop more specific and “personal” diagnostic and therapeutic approaches.
Claudins (CLDNs) are tight junction (TJ) proteins that serve an intercellular adhesion function. The aberrant expression of individual claudins is well documented in different stages of various human cancers [12]. In addition, some claudins were proven to be useful as biomarkers [13, 14].
The insulin-like growth factor (IGF) axis attracts increasing attention since it is involved in several stages of cancer [15, 16, 17], and for its vital role in regulating cell survival and growth [18, 19] as well as the possible use of constituents of this axis as tumor and/or metastasis markers, which is becoming the focus of increasing research activities.
The insulin-like growth factors IGF-I and –II orchestrate their roles through the interaction with other members in this system, namely their receptors IGF-IR and -IIR, their binding proteins (IGFBPs) and the IGFBP proteases including matrix metalloproteinases (MMPs), cathepsins, and kallikreins [20]. Type I receptor mediates the growth promoting effects of IGFs [21], which are further modulated by 6 binding proteins (IGFBPs 1–6) with high affinity for IGFs [22] as well as at least 4 IGFBPs with low affinity [23, 24], also known as IGFBP-related proteins (IGFBP-rp-1-4).
Based on the observation that the increased expression of IGFBPs attenuates the proliferative and anti-apoptotic effects of IGFs, they have been long considered as tumor suppressors, mostly due to their IGF-dependent roles. Interestingly, however, in addition to these IGF-dependent actions, IGFBPs were found to exert IGF-independent effects, as was reported for IGFBP1 [25], IGFBP3 [26, 27, 28], IGFBP5 [29] and IGFBP7 (or IGFBP-rp1) [30, 31, 32].
In this report, we have used the technique of cancer cell reisolation from rat liver, which permitted to monitor for the first time the expression profile of numerous candidate genes in a time-dependent manner. Based on these results we summarize our knowledge on claudins and IGFBP members and delineate their potential as tumor and/or metastasis markers.
2. Results
2.1 Modulation of selected genes in reisolated CC531 tumor cells
For identifying genes, which enable tumor cells to metastasize and colonize the liver, the CC531 cells were reisolated from rats, which had been implanted intraportally with these tumor cells. After various periods following tumor cell implantation, the CC531 cells were reisolated with a specific technique [33]. In subsequent experiments, these cells were used for mRNA and protein isolation and the mRNA screened by cDNA microarray and RT-PCR, and the proteins by Western blot.
As shown in Table 1, the microarray analysis revealed a significantly increased expression of insulin like growth factor binding proteins (
2.2 Effects of genes’ knockdown in colorectal cancer cells
To investigate the knockdown effect(s) of each gene on various functions of colorectal cancer cell lines, siRNA experiments for transient knockdown were performed.
Colorectal cancer cells (CC531, Caco2 or SW480) cultured in 6-well-plates were transfected with specific siRNA (200 nM) or negative control using Lipofectamine 2000 (Invitrogen) following the manufacturer’s instructions. The cells were harvested at 24, 48 and 72 h after treatment.
As shown for overview in Table 2 (for details see Figures 2–4), knockdown of
Target gene | Cell proliferation | Cell migration | Colony formation |
---|---|---|---|
↓ | ↓ | ↓ | |
↓ | ns | ↓ | |
ns | ↑ | ↓ | |
ns | ↑ | ↓ |
Interestingly, different effects were noticed after
3. Discussion
“If it were not for the great variability among individuals, medicine might as well be a science and not an art“ with these words the famous Canadian physician Sir William Osler anticipated and acknowledged the concept of personalized medicine since the nineteenth century.
Personalized medicine aims to optimize and tailor preventive and therapeutic approaches in favor of the best outcome for each patient, by using genetics, proteomics, and biological information, including biomarkers [34]. It attempts to sub-categorize patients into different groups according to their “molecular make up”, i.e. using biomarkers.
Here we have used a rat model of liver metastasis to identify genes with importance for organ colonization, which could be used as biomarkers or therapeutic targets.
The selected model for this experiment is characterized by a defined metastatic proliferation-onset in rat liver after intra-portal inoculation of CC531 cancer cells. Consequently, this allows exploring the chronological modulation of gene expression, from the very beginning of cancer cell homing into the liver to their final colonization of the whole organ. The technique of cancer cell reisolation from rat liver permitted, for the first time, monitoring the expression profile of numerous candidate genes from the whole genome in a time-dependent manner.
The initial observation of these studies was based on cDNA microarray analysis of CC531 colon cancer cells, which allowed selecting candidates from gene families with significant up- or down-regulation. These candidates were further analyzed by different
We have been focusing on the detection and evaluation of biomarkers for the last decades [33, 35, 36, 37, 38, 39, 40, 41, 42, 43]. From the several gene groups, which were highly modulated in expression during liver colonization we focused on two families. Both were characterized by dramatic initial changes in expression, with claudins being down modulated and IGFBPs being up-regulated.
The first group (claudins; CLDNs) form the structural backbone of tight junctions (TJs), one type of cell-cell adhesion, and comprise at least 27 members of integral transmembrane proteins ranging in size from 20 to 34 kDa [12]. In recent years, the up- or down-modulated expression of several claudins has been associated with the progression of various cancers in humans, even in a tissue specific manner [12].
Interestingly, individual claudins are being used as therapeutic targets [44, 45] as well as diagnostic biomarkers [13, 14], making them a very interesting molecule to be investigated and characterized.
The second group of insulin like growth factor binding proteins (IGFBPs) differs in its importance from claudins, as they belong to the insulin-like growth factor (IGF) axis, which has a vital role in regulating cell survival and growth and is involved in several stages of cancer.
The expression of the two IGFBP genes, igfbp3 and igfbp7, was intensely upregulated at the beginning of liver colonization (days 3 and 6 after tumor implantation). Subsequently, however, this increased expression returned gradually to normal, hence we assume that the up-regulation of IGFBPs is essential for dissemination and homing of tumor cells into the liver during early metastasis formation. This strongly suggests that the tumor/metastasis microenvironment has a crucial impact on the regulation of igfbp3 and igfbp7. Furthermore, these results along with previous studies [28, 46, 47, 48] show that the balanced expression of IGFBP3 and IGFBP7 is very essential for several cellular functions as both, up- and down-regulation of these two genes were related to malignant properties. This further suggests that the aberrant expression of these two genes can be an early indicator of CRC progression.
Further verification of their value arises from studies that demonstrate involvement of the IGF axis in several stages of cancer and for its vital role in regulating cell and tissue survival, growth and differentiation [18, 19]. In addition, the possible use of constituents of this axis as tumor and/or metastasis markers is becoming the focus of increasing research activities [49, 50, 51]. Most
IGFBP3 is well known in the literature: It is the predominant IGFBP in plasma, hence plays a crucial role in regulating the bioavailability of plasma IGFs, and it is expressed locally in most tissues including the intestine [55]. Additionally, IGFBP3 induces apoptosis and inhibits proliferation in human colon [56], prostate [57], breast [58], and lung cancer cells [26]
In contrast to IGFBP3, the regulation and functions of IGFBP7 are less investigated. This gene was originally cloned as a gene, which is down-regulated in meningioma cell lines [68]. IGFBP7 is usually expressed by colonic mucosa [65], however both, up- and down-regulation patterns were recorded in the context of cancer [46, 47]. Lately, it was shown that IGFBP7 is a direct p53 target and the DNA methylation mediated-epigenetic silencing of IGFBP7 was associated with the absence of p53 mutations in CRC [30]. Until now,
As also known from the literature, several members of the IGF axis were found to be prognostic markers for various tumor types, including IGFBP5 and IGFBP7 for nonsmall cell lung cancer (NSCLC) [72], IGFBP2 and IGFBP3 as compensatory biomarkers for CA19-9 in early-stage pancreatic cancer [51], IGF-1 for metastatic uveal melanoma [49] and IGF-IR for glioblastoma [50]. In addition, a recent study showed that certain single nucleotide polymorphisms (SNPs) in IGF1R and IGF2R were associated (positively or inversely, respectively) with adenomas in Caucasian, but not in African American CRC patients [73]. Similarly, specific SNPs in the IGF-l gene were suggested as risk assessment markers of gastrointestinal cancers [74]. All these studies emphasize the crucial role of the patient’s genetic background in tailoring the therapeutic approach to fit the “size” of this particular patient.
With regard to using the above family members as prognostic marker, the past experience should be considered. Contrasting with our expectations and reports from the literature, no significant correlation was found between the increased expression levels of four known tumor progression-associated genes (Opn, Tgf-β, Mmp-2 and Cox-2) and the prognostic value of these genes in CRC patients [43]. This raises an important to answer question, i.e. what would be the best procedure to apply personalized medicine effectively and reliably? Could it be a minimum number of (bio)-markers for each cancer type to be tested, or should it be a complex approach as high throughput genome sequencing, as it is increasingly performed?
Ideally, a few markers would be better suited regarding costs of analysis and time until a patient can benefit from the results. However, there are currently only few markers which succeeded to be applied accordingly. Therefore, even more methods are being approached to assess the specific changes inherent to the full genome.
4. Conclusion and perspectives
Here we show a new high throughput approach in exploring genes relevant to CRC progression in terms of liver metastasis. Our method has yielded initial results related to the importance of claudins and IGFBP in liver colonization. Nevertheless, we reason that other genes, which result from this model, might be even more valuable. For instance, one of the very important and interesting gene families that resulted from this model, which is extensively investigated, is the endothelin system with all its components (endothelins, their converting enzymes and their receptors). Several members of this system could prove useful as tumor/metastasis markers. Future experiments will show whether such a functional model can compete or complement, at least partially, other techniques, including whole genome sequencing.
References
- 1.
Arnold M, Sierra MS, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global patterns and trends in colorectal cancer incidence and mortality. Gut. 2017; 4 :683-691. DOI: 10.1136/gutjnl-2015-310912 - 2.
Valderrama-Trevino AI, Barrera-Mera B, Ceballos-Villalva JC, Montalvo-Jave EE. Hepatic metastasis from colorectal cancer. Euroasian Journal of Hepato-Gastroenterology. 2017; 2 :166-175. DOI: 10.5005/jp-journals-10018-1241 - 3.
Bismuth H, Adam R, Levi F, Farabos C, Waechter F, Castaing D, et al. Resection of nonresectable liver metastases from colorectal cancer after neoadjuvant chemotherapy. Annals of Surgery. 1996; 4 :509-520. discussion 520-502 - 4.
Fong Y, Fortner J, Sun RL, Brennan MF, Blumgart LH. Clinical score for predicting recurrence after hepatic resection for metastatic colorectal cancer: Analysis of 1001 consecutive cases. Annals of Surgery. 1999; 3 :309-318. discussion 318-321 - 5.
Wittmer A, Khazaie K, Berger MR. Quantitative detection of lac-Z-transfected CC531 colon carcinoma cells in an orthotopic rat liver metastasis model. Clinical & Experimental Metastasis. 1999; 5 :369-376 - 6.
Seelig MH, Leible M, Sanger J, Berger MR. Chemoembolization of rat liver metastasis with microspheres and gemcitabine followed by evaluation of tumor cell load by chemiluminescence. Oncology Reports. 2004; 5 :1107-1113 - 7.
Eyol E, Boleij A, Taylor RR, Lewis AL, Berger MR. Chemoembolisation of rat colorectal liver metastases with drug eluting beads loaded with irinotecan or doxorubicin. Clinical & Experimental Metastasis. 2008; 3 :273-282 - 8.
Marinelli A, van Dierendonck JH, van Brakel GM, Irth H, Kuppen PJ, Tjaden UR, et al. Increasing the effective concentration of melphalan in experimental rat liver tumours: Comparison of isolated liver perfusion and hepatic artery infusion. British Journal of Cancer. 1991; 6 :1069-1075 - 9.
Oldenburg J, Begg AC, van Vugt MJ, Ruevekamp M, Schornagel JH, Pinedo HM, et al. Characterization of resistance mechanisms to cis-diamminedichloroplatinum(II) in three sublines of the CC531 colon adenocarcinoma cell line in vitro. Cancer Research. 1994; 2 :487-493 - 10.
Veenhuizen RB, Marijnissen JP, Kenemans P, Ruevekamp-Helmers MC, t Mannetje LW, Helmerhorst TJ, et al. Intraperitoneal photodynamic therapy of the rat CC531 adenocarcinoma. British Journal of Cancer. 1996; 11 :1387-1392 - 11.
Biomarkers Definitions Working Group. Biomarkers and surrogate endpoints: Preferred definitions and conceptual framework. Clinical Pharmacology & Therapeutics. 2001; 3 :89-95. DOI: 10.1067/mcp.2001.113989 - 12.
Osanai M, Takasawa A, Murata M, Sawada N. Claudins in cancer: Bench to bedside. Pflügers Archiv/European Journal of Physiology. 2017; 1 :55-67. DOI: 10.1007/s00424-016-1877-7 - 13.
Benczik M, Galamb A, Koiss R, Kovacs A, Jaray B, Szekely T, et al. Claudin-1 as a biomarker of cervical cytology and histology. Pathology Oncology Research. 2016; 1 :179-188. DOI: 10.1007/s12253-015-9990-z - 14.
Szabo I, Kiss A, Schaff Z, Sobel G. Claudins as diagnostic and prognostic markers in gynecological cancer. Histology and Histopathology. 2009; 12 :1607-1615. DOI: 10.14670/HH-24.1607 - 15.
Chhabra Y, Waters MJ, Brooks AJ. Role of the growth hormone-IGF-1 axis in cancer. Expert Review of Endocrinology and Metabolism. 2011; 1 :71-84. DOI: 10.1586/eem.10.73 - 16.
Samani AA, Yakar S, LeRoith D, Brodt P. The role of the IGF system in cancer growth and metastasis: Overview and recent insights. Endocrine Reviews. 2007; 1 :20-47. DOI: 10.1210/er.2006-0001 - 17.
LeRoith D, Roberts CT Jr. The insulin-like growth factor system and cancer. Cancer Letters. 2003; 2 :127-137. DOI: 10.1016/S0304-3835(03)00159-9 - 18.
Stewart CE, Rotwein P. Growth, differentiation, and survival: Multiple physiological functions for insulin-like growth factors. Physiological Reviews. 1996; 4 :1005-1026 - 19.
Bustin SA, Jenkins PJ. The growth hormone-insulin-like growth factor-I axis and colorectal cancer. Trends in Molecular Medicine. 2001; 10 :447-454. DOI: S1471-4914(01)02104 - 20.
Rajah R, Katz L, Nunn S, Solberg P, Beers T, Cohen P. Insulin-like growth factor binding protein (IGFBP) proteases: Functional regulators of cell growth. Progress in Growth Factor Research. 1995; 2 (4):273-284 - 21.
Grimberg A, Cohen P. Role of insulin-like growth factors and their binding proteins in growth control and carcinogenesis. Journal of Cellular Physiology. 2000; 1 :1-9 - 22.
Baxter RC, Binoux MA, Clemmons DR, Conover CA, Drop SL, Holly JM, et al. Recommendations for nomenclature of the insulin-like growth factor binding protein superfamily. The Journal of Clinical Endocrinology and Metabolism. 1998; 9 :3213 - 23.
Hwa V, Oh Y, Rosenfeld RG. The insulin-like growth factor-binding protein (IGFBP) superfamily. Endocrine Reviews. 1999; 6 :761-787 - 24.
Kostecka Y, Blahovec J. Insulin-like growth factor binding proteins and their functions (minireview). Endocrine Regulations. 1999; 2 :90-94 - 25.
Peterkofsky B, Gosiewska A, Wilson S, Kim YR. Phosphorylation of rat insulin-like growth factor binding protein-1 does not affect its biological properties. Archives of Biochemistry and Biophysics. 1998; 1 :101-110 - 26.
Lee KW, Cohen P. Nuclear effects: Unexpected intracellular actions of insulin-like growth factor binding protein-3. The Journal of Endocrinology. 2002; 1 :33-40. DOI: 10.1677/joe.0.1750033 - 27.
Baxter RC. Signalling pathways involved in antiproliferative effects of IGFBP-3: A review. Molecular Pathology. 2001; 3 :145-148 - 28.
Kansra S, Ewton DZ, Wang J, Friedman E. IGFBP-3 mediates TGF beta 1 proliferative response in colon cancer cells. International Journal of Cancer. 2000; 3 :373-378 - 29.
Nam TJ, Busby W Jr, Clemmons DR. Insulin-like growth factor binding protein-5 binds to plasminogen activator inhibitor-I. Endocrinology. 1997; 7 :2972-2978 - 30.
Suzuki H, Igarashi S, Nojima M, Maruyama R, Yamamoto E, Kai M, et al. IGFBP7 is a p53-responsive gene specifically silenced in colorectal cancer with CpG island methylator phenotype. Carcinogenesis. 2010; 3 :342-349. DOI: 10.1093/carcin/bgp179 - 31.
Ruan WJ, Lin J, Xu EP, Xu FY, Ma Y, Deng H, et al. IGFBP7 plays a potential tumor suppressor role against colorectal carcinogenesis with its expression associated with DNA hypomethylation of exon 1. Journal of Zhejiang University. Science. B. 2006; 11 :929-932. DOI: 10.1631/jzus.2006.B0929 - 32.
Ahmed S, Yamamoto K, Sato Y, Ogawa T, Herrmann A, Higashi S, et al. Proteolytic processing of IGFBP-related protein-1 (TAF/angiomodulin/mac25) modulates its biological activity. Biochemical and Biophysical Research Communications. 2003; 2 :612-618. DOI: 10.1016/j.bbrc.2003.09.058 - 33.
Georges R, Bergmann F, Hamdi H, Zepp M, Eyol E, Hielscher T, et al. Sequential biphasic changes in claudin1 and claudin4 expression are correlated to colorectal cancer progression and liver metastasis. Journal of Cellular and Molecular Medicine. 2012; 2 :260-272. DOI: 10.1111/j.1582-4934.2011.01289.x - 34.
Schleidgen S, Klingler C, Bertram T, Rogowski WH, Marckmann G. What is personalized medicine: Sharpening a vague term based on a systematic literature review. BMC Medical Ethics. 2013; 55 . DOI: 10.1186/1472-6939-14-55 - 35.
Schimanski CC, Linnemann U, Berger MR. Sensitive detection of K-ras mutations augments diagnosis of colorectal cancer metastases in the liver. Cancer Research. 1999; 20 :5169-5175 - 36.
Shao L, Huang Q, Luo M, Lai M. Detection of the differentially expressed gene IGF-binding protein-related protein-1 and analysis of its relationship to fasting glucose in Chinese colorectal cancer patients. Endocrine-Related Cancer. 2004; 1 :141-148. DOI: 10.1677/erc.0.0110141 - 37.
Conzelmann M, Linnemann U, Berger MR. K-ras codon 12 and 13 mutations are correlated with differential patterns of tumor cell dissemination in colorectal cancer patients. International Journal of Oncology. 2004; 6 :1537-1544 - 38.
Schimanski CC, Zimmermann T, Schmidtmann I, Gockel I, Lang H, Galle PR, et al. K-ras mutation status correlates with the expression of VEGFR1, VEGFR2, and PDGFRalpha in colorectal cancer. International Journal of Colorectal Disease. 2010; 2 :181-186. DOI: 10.1007/s00384-009-0843-7 - 39.
Schimanski CC, Mohler M, Zimmermann T, Gockel I, Lang H, Arbogast R, et al. Detection of hepatic micrometastases in the context of adjuvant chemotherapy and surgery for hepatic metastases. Deutsche Medizinische Wochenschrift. 2010; 21 :1078-1082. DOI: 10.1055/s-0030-1253703 - 40.
Georges R, Adwan H, Zhivkova M, Eyol E, Bergmann F, Berger MR. Regulation of osteopontin and related proteins in rat CC531 colorectal cancer cells. International Journal of Oncology. 2010; 2 :249-256 - 41.
Schimanski CC, Moehler M, Gockel I, Zimmermann T, Lang H, Galle PR, et al. Expression of chemokine receptor CCR5 correlates with the presence of hepatic molecular metastases in K-ras positive human colorectal cancer. Journal of Cancer Research and Clinical Oncology. 2011; 7 :1139-1145. DOI: 10.1007/s00432-011-0980-6 - 42.
Allami RH, Graf C, Martchenko K, Voss B, Becker M, Berger MR, et al. Analysis of the expression of SDF-1 splicing variants in human colorectal cancer and normal mucosa tissues. Oncology Letters. 2016; 3 :1873-1878. DOI: 10.3892/ol.2016.4139 - 43.
Uhlmann ME, Georgieva M, Sill M, Linnemann U, Berger MR. Prognostic value of tumor progression-related gene expression in colorectal cancer patients. Journal of Cancer Research and Clinical Oncology. 2012; 10 :1631-1640. DOI: 10.1007/s00432-012-1238-7 - 44.
Hashimoto Y, Fukasawa M, Kuniyasu H, Yagi K, Kondoh M. Claudin-targeted drug development using anti-claudin monoclonal antibodies to treat hepatitis and cancer. Annals of the New York Academy of Sciences. 2017; 1 :5-16. DOI: 10.1111/nyas.13337 - 45.
Morin PJ. Claudin proteins in human cancer: Promising new targets for diagnosis and therapy. Cancer Research. 2005; 21 :9603-9606. DOI: 10.1158/0008-5472.CAN-05-2782 - 46.
How HK, Yeoh A, Quah TC, Oh Y, Rosenfeld RG, Lee KO. Insulin-like growth factor binding proteins (IGFBPs) and IGFBP-related protein 1-levels in cerebrospinal fluid of children with acute lymphoblastic leukemia. The Journal of Clinical Endocrinology and Metabolism. 1999; 4 :1283-1287 - 47.
Komatsu S, Okazaki Y, Tateno M, Kawai J, Konno H, Kusakabe M, et al. Methylation and downregulated expression of mac25/insulin-like growth factor binding protein-7 is associated with liver tumorigenesis in SV40T/t antigen transgenic mice, screened by restriction landmark genomic scanning for methylation (RLGS-M). Biochemical and Biophysical Research Communications. 2000; 1 :109-117 - 48.
Leibowitz BJ, Cohick WS. Endogenous IGFBP-3 is required for both growth factor-stimulated cell proliferation and cytokine-induced apoptosis in mammary epithelial cells. Journal of Cellular Physiology. 2009; 1 :182-188. DOI: 10.1002/jcp.21748 - 49.
Frenkel S, Zloto O, Pe'er J, Barak V. Insulin-like growth factor-1 as a predictive biomarker for metastatic uveal melanoma in humans. Investigative Ophthalmology & Visual Science. 2013; 1 :490-493. DOI: 10.1167/iovs.12-10228 - 50.
Maris C, D'Haene N, Trepant AL, Le Mercier M, Sauvage S, Allard J, et al. IGF-IR: A new prognostic biomarker for human glioblastoma. British Journal of Cancer. 2015; 5 :729-737. DOI: 10.1038/bjc.2015.242 - 51.
Yoneyama T, Ohtsuki S, Honda K, Kobayashi M, Iwasaki M, Uchida Y, et al. Identification of IGFBP2 and IGFBP3 as compensatory biomarkers for CA19-9 in early-stage pancreatic cancer using a combination of antibody-based and LC-MS/MS-based proteomics. PLoS One. 2016; 8 :e0161009. DOI: 10.1371/journal.pone.0161009 - 52.
Ma Y, Lu B, Ruan W, Wang H, Lin J, Hu H, et al. Tumor suppressor gene insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1) induces senescence-like growth arrest in colorectal cancer cells. Experimental and Molecular Pathology. 2008; 2 :141-145. DOI: 10.1016/j.yexmp.2008.04.005 - 53.
Mohan S, Baylink DJ. IGF-binding proteins are multifunctional and act via IGF-dependent and -independent mechanisms. The Journal of Endocrinology. 2002; 1 :19-31. DOI: 10.1677/joe.0.1750019 - 54.
Jiang W, Xiang C, Cazacu S, Brodie C, Mikkelsen T. Insulin-like growth factor binding protein 7 mediates glioma cell growth and migration. Neoplasia. 2008; 12 :1335-1342. DOI: 10.1593/neo.08694 - 55.
Winesett DE, Ulshen MH, Hoyt EC, Mohapatra NK, Fuller CR, Lund PK. Regulation and localization of the insulin-like growth factor system in small bowel during altered nutrient status. The American Journal of Physiology. 1995; 4 (Pt 1):G631-G640 - 56.
Williams AC, Collard TJ, Perks CM, Newcomb P, Moorghen M, Holly JM, et al. Increased p53-dependent apoptosis by the insulin-like growth factor binding protein IGFBP-3 in human colonic adenoma-derived cells. Cancer Research. 2000; 1 :22-27 - 57.
Hong J, Zhang G, Dong F, Rechler MM. Insulin-like growth factor (IGF)-binding protein-3 mutants that do not bind IGF-I or IGF-II stimulate apoptosis in human prostate cancer cells. The Journal of Biological Chemistry. 2002; 12 :10489-10497. DOI: 10.1074/jbc.M109604200 - 58.
Gill ZP, Perks CM, Newcomb PV, Holly JM. Insulin-like growth factor-binding protein (IGFBP-3) predisposes breast cancer cells to programmed cell death in a non-IGF-dependent manner. The Journal of Biological Chemistry. 1997; 41 :25602-25607. DOI: 10.1074/jbc.272.41.25602 - 59.
Kirman I, Poltoratskaia N, Sylla P, Whelan RL. Insulin-like growth factor-binding protein 3 inhibits growth of experimental colocarcinoma. Surgery. 2004; 2 :205-209. DOI: 10.1016/j.surg.2004.04.020 - 60.
Ogino S, Meyerhardt JA, Kawasaki T, Clark JW, Ryan DP, Kulke MH, et al. CpG island methylation, response to combination chemotherapy, and patient survival in advanced microsatellite stable colorectal carcinoma. Virchows Archiv. 2007; 5 :529-537. DOI: 10.1007/s00428-007-0398-3 - 61.
Chan JM, Stampfer MJ, Ma J, Gann P, Gaziano JM, Pollak M, et al. Insulin-like growth factor-I (IGF-I) and IGF binding protein-3 as predictors of advanced-stage prostate cancer. Journal of the National Cancer Institute. 2002; 14 :1099-1106 - 62.
Krajcik RA, Borofsky ND, Massardo S, Orentreich N. Insulin-like growth factor I (IGF-I), IGF-binding proteins, and breast cancer. Cancer Epidemiology, Biomarkers & Prevention. 2002; 12 :1566-1573 - 63.
Palmqvist R, Hallmans G, Rinaldi S, Biessy C, Stenling R, Riboli E, et al. Plasma insulin-like growth factor 1, insulin-like growth factor binding protein 3, and risk of colorectal cancer: A prospective study in northern Sweden. Gut. 2002; 5 :642-646 - 64.
DeLellis K, Rinaldi S, Kaaks RJ, Kolonel LN, Henderson B, Le Marchand L. Dietary and lifestyle correlates of plasma insulin-like growth factor-I (IGF-I) and IGF binding protein-3 (IGFBP-3): The multiethnic cohort. Cancer Epidemiology, Biomarkers & Prevention. 2004; 9 :1444-1451 - 65.
Durai R, Yang W, Gupta S, Seifalian AM, Winslet MC. The role of the insulin-like growth factor system in colorectal cancer: Review of current knowledge. International Journal of Colorectal Disease. 2005; 3 . DOI: 203-220. 10.1007/s00384-004-0675-4 - 66.
Serel TA, Kecelioglu M. Serum insulin-like growth factor is not a useful marker of prostate cancer. BJU International. 2000; 4 :559-560 - 67.
Kaaks R, Toniolo P, Akhmedkhanov A, Lukanova A, Biessy C, Dechaud H, et al. Serum C-peptide, insulin-like growth factor (IGF)-I, IGF-binding proteins, and colorectal cancer risk in women. Journal of the National Cancer Institute. 2000; 19 :1592-1600 - 68.
Murphy M, Pykett MJ, Harnish P, Zang KD, George DL. Identification and characterization of genes differentially expressed in meningiomas. Cell Growth & Differentiation. 1993; 9 :715-722 - 69.
Kato MV. A secreted tumor-suppressor, mac25, with activin-binding activity. Molecular Medicine. 2000; 2 :126-135 - 70.
Wilson HM, Birnbaum RS, Poot M, Quinn LS, Swisshelm K. Insulin-like growth factor binding protein-related protein 1 inhibits proliferation of MCF-7 breast cancer cells via a senescence-like mechanism. Cell Growth & Differentiation. 2002; 5 :205-213 - 71.
Adachi Y, Itoh F, Yamamoto H, Arimura Y, Kikkawa-Okabe Y, Miyazaki K, et al. Expression of angiomodulin (tumor-derived adhesion factor/mac25) in invading tumor cells correlates with poor prognosis in human colorectal cancer. International Journal of Cancer. 2001; 4 :216-222 - 72.
Shersher DD, Vercillo MS, Fhied C, Basu S, Rouhi O, Mahon B, et al. Biomarkers of the insulin-like growth factor pathway predict progression and outcome in lung cancer. The Annals of Thoracic Surgery. 2011; 5 :1805-1811. DOI: 10.1016/j.athoracsur.2011.06.058. discussion 1811 - 73.
Ochs-Balcom HM, Vaughn CB, Nie J, Chen Z, Thompson CL, Parekh N, et al. Racial differences in the association of insulin-like growth factor pathway and colorectal adenoma risk. Cancer Causes & Control. 2014; 2 :161-170. DOI: 10.1007/s10552-013-0318-6 - 74.
Ong J, Salomon J, te Morsche RH, Roelofs HM, Witteman BJ, Dura P, et al. Polymorphisms in the insulin-like growth factor axis are associated with gastrointestinal cancer. PLoS One. 2014; 3 :e90916. DOI: 10.1371/journal.pone.0090916