Copy number changes of CRISP3 in oral squamous cell carcinoma

  • Authors:
    • Wen-Chang Ko
    • Keisuke Sugahara
    • Takumi Sakuma
    • Ching-Yu Yen
    • Shyun-Yeu Liu
    • Gwo-An Liaw
    • Takahiko Shibahara
  • View Affiliations

  • Published online on: September 9, 2011     https://doi.org/10.3892/ol.2011.418
  • Pages: 75-81
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The aim of this study was to identify tumor suppressor genes (TSGs) in oral squamous cell carcinoma (OSCC) using whole-genome analysis of microarray technology and real-time quantitative polymerase chain reaction (QPCR). We applied whole-genome analysis of TSGs in the specimens from 3 patients of OSCC by microarray technology. A total of 11 genes, CRISP3, SCGB3A1, AGR2, PIP, C20orf114, TFF1, STATH, AZGP1, MUC7, DMBT1 and LOC389429, were found to be down-regulated, and 2, matrix metallopeptidase (MMP) 1 and MMP3, were found to be up-regulated in the 3 OSCC patients using microarray technology. In this study, we selected the CRISP3 gene. CRISP3 belongs to the cystein-rich secretary protein gene family in chromosome 6p12.3. CRISP3 has been found in the salivary gland, spleen and prostate gland and is a prominent biomarker in the gene expression of prostate cancer. Down-regulation of this gene was previously observed in OSCC. No studies examining the DNA copy number of CRISP3 in detail exist. We analyzed the DNA copy number of CRISP3 in 5 OSCC-derived cell lines (SAS, Ca9-22, KON, HSC2 and HSC4) and 60 OSCC tissues by real-time QPCR. The DNA copy number loss of CRISP3 was observed in 2 of the 5 OSCC-derived cell lines (SAS, HSC2) and in 24 of 60 patients (40.0%) using real-time QPCR. A significant statistical correlation between the copy number loss and gender and T classification was observed. These results indicate that the inactivation of CRISP3 is an early event in OSCC, since the T1/T2 classification is correlated with DNA copy number loss of CRISP3, whereas T3/T4 classification is not. We conclude that CRISP3 may be involved in the carcinogenesis of OSCC.

Introduction

Oral squamous cell carcinoma (OSCC) is a common malignancy that affects 300,000 individuals per year worldwide (1). A number of etiologic factors have been implicated in the development of OSCCs, such as the use of tobacco, alcohol or betel nut chewing, human papillomavirus infection, and the presence of incompatible prosthetic materials. Tobacco is a major risk factor for the development of OSCC and overall risk of OSCC among smokers is 7–10 times higher than that of non-smokers (24).

The primary treatment for management of OSCC is surgical intervention. Despite considerable advances in the treatment of OSCC over the past two decades, overall disease outcomes have only modestly improved (5). Local tumor recurrence affects approximately 60% of patients and metastasis develops in 15–25% (6). The prevention and management of this disease is likely to greatly benefit from the identification of molecular markers and targets (7,8).

However, little is known about the molecular basis of OSCC compared with other malignancies. Molecular alterations in a number of oncogenes and tumor suppressor genes (TSGs) associated with the development of OSCC may be significant clues with which to address these problems (9,10).

Inactivation of TSG is considered to be associated with carcinogenesis, and alterations in TSGs are accepted to be critical events in the multi-step process leading toward the development of cancer. Loss of chromosome 3p22-24 is a common early genetic event in OSCC (11). Specific regions of arm harbor candidate tumor suppressor genes including FHIT and RSSF1A. It is generally accepted that the transformation of normal tissue into malignant tissue follows an accumulation of genetic changes in the TSGs and oncogenes (9). High throughput investigation into the molecular characteristics of OSCC has mainly utilized microarray technology to search for gene expression profiles associated with disease and disease outcome.

In the present study, microarray technology was applied to screen novel TSGs in OSCC patients, and we selected candidate gene CRISP3. CRISP3 DNA copy number was evaluated by real-time QPCR in 5 OSCC-derived cell lines and 60 primary OSCC samples. Human CRISP3 is also likely to be involved in the pathogenesis of prostate cancer, where CRISP3 expression is significantly upregulated (12). CRISP3 is secreted and can be detected in human tissue fluids including saliva, sweat, blood and seminal plasma, rendering it an ideal candidate biomarker for pathophysiological conditions, including ectopic pregnancy (1214). Ye et al reported the expression of CRISP3 in OSCC using microarray technology (15). Therefore, the purpose of the present study was to assess the role of CRISP3 in OSCC.

Materials and methods

Cells

The 5 human OSCC-derived cell lines used in this study were SAS, Ca9-22, KON, HSC2, and HSC4 (Human Science Research Resources Bank, Osaka, Japan). SAS was from male tongue, Ca9-22 from male gingiva, KON from male oral floor, HSC2 from male mouth and HSC4 from male tongue. The cell lines were maintained at 37°C (humidified atmosphere 5% CO2/95% air) in 150×200 mm tissue culture dishes (Nunc, Roskilde, Denmark) and cultured in Dulbecco’s modified Eagle’s medium F-12 HAM (Sigma, St. Louis, MO, USA) with 10% fetal bovine serum (Sigma) plus 50 U/ml penicillin and streptomycin.

Normal oral keratinocyte (NOKs) strains from two patients who had undergone dental surgery served as the controls, and the patients provided written informed consent prior to the start of the study. The normal oral specimens were washed in Dulbecco’s phosphate-buffered saline (PBS) (Sigma) and then placed overnight in 0.25% trypsin-EDTA solution (Sigma) at 4°C. After the epithelial tissue was separated from the connective tissue, it was disaggregated by incubation in 0.25% trypsin-EDTA solution for 15 min with gentle pipetting at 37°C. Isolated epithelial cells were then seeded into Collagen I Cellware 60-mm dish biocoat cell environments (Becton Dickinson Labware, Bedford, MA, USA) and cultured in Keratinocyte Basal Medium-2 (Cambrex, Walkersville, MD, USA) with 0.4% bovine pituitary extract, 0.1% human epidermal growth factor, 0.1% insulin, 0.1% hydrocortisone, 0.1% transferrin, 0.1% epinephrine and 0.1% GA-1000 (Cambrex) (16).

Patient characteristics

A total of 60 patients with OSCC were included in the present study. Surgical resection of primary tumors and marginal normal tissues from all patients was performed at the Hospital of Chimei, Tainan, Taiwan, and the Hospital of Tokyo Dental College, Chiba, Japan, between July 1999 and September 2009. Written informed consent was obtained from all patients and the study approved by the ethics committees of the Hospital of Chimei and the Tokyo Dental College, ethical clearance number 205. Informed consent was obtained from each patient prior to surgical resection.

Staging of tumors was performed according to the International Union Against Cancer TNM staging system (17). Cervical LNM during the 12-month follow-up period was evaluated by computed tomography and magnetic resonance imaging. In case of a positive signal, metastasis was further confirmed by histopathological examination of the resected tissues. Patients not exhibiting any cervical LNM for 12 months after surgery were considered metastasis-free. Patients with distant metastasis at the time of clinical examination or those receiving preventive radiotherapy or chemotherapy were excluded from this study. Detailed patient characteristics are shown in Table I.

Table I

Clinical characteristics of 60 OSCC patients.

Table I

Clinical characteristics of 60 OSCC patients.

CaseGenderAgeEthnic groupTumor siteTNStagepNTobaccoAlcohol
1M43TaiwaneseBuccal mucosa30III++
2M66TaiwaneseLower gingival20II++
3M31TaiwaneseTongue20II++
4M55TaiwaneseBuccal mucosa10I++
5M37TaiwaneseTongue30III++
6M58TaiwaneseBuccal mucosa10I++
7F71TaiwaneseUpper gingival20II
8M43TaiwaneseOral floor20II++
9M39TaiwaneseBuccal mucosa20II++
10M56TaiwaneseTongue10I++
11M49TaiwaneseBuccal mucosa30III++
12M72TaiwaneseTongue11III+++
13M51TaiwaneseLower gingival40IV++
14M55TaiwaneseTongue22IV+
15M59TaiwaneseBuccal mucosa31III+
16M54TaiwaneseBuccal mucosa10I++
17M68TaiwaneseBuccal mucosa20II+
18M33JapaneseTongue21III++
19M55JapaneseOral floor32cIV+++
20M74JapaneseTongue21III++
21M54JapaneseBuccal mucosa42cIV++
22M64JapaneseLower gingival42aIV++
23M53JapaneseTongue21III++
24F70JapaneseBuccal mucosa31III+
25M44JapaneseTongue10I+
26F50JapaneseUpper gingival10I+
27M36JapaneseUpper gingival20II+
28F37JapaneseTongue22cIV
29M47JapaneseTongue20II++
30M43JapaneseTongue10I+
31M57JapaneseTongue22bIV++
32M52JapaneseTongue10I++
33F58JapaneseTongue20II
34M59JapaneseUpper gingival31III+
35M62JapaneseTongue10I++
36F70JapaneseUpper gingival21III
37M82JapaneseUpper gingival30III+
38F69JapaneseBuccal mucosa20II
39M80JapaneseLower gingival22bIV+
40M48JapaneseTongue10I+
41M70JapaneseLower gingival21III++
42M49JapaneseLower gingival22bIV
43M85JapaneseTongue21III
44M71JapaneseLower gingival41IV+
45F73JapaneseTongue10I
46F58JapaneseLower gingival30III
47M41JapaneseTongue32bIV++
48M58JapaneseTongue11III+++
49F66JapaneseBuccal mucosa21III+
50M63JapaneseTongue10I++
51F60JapaneseLower gingival20II
52M52JapaneseTongue20II++
53F72JapaneseUpper gingival22aIV
54F63JapaneseLower gingival42bIV
55F66JapaneseBuccal mucosa10I
56M66JapaneseOral floor42cIV+++
57M66JapaneseLower gingival42bIV+
58M76JapaneseOral floor21III++
59M77JapaneseOral floor21III+
60F69JapaneseLower gingival42bIV+
Primary tumor samples

Resected primary tumor tissues were divided into two sections. Of these, one section was frozen immediately in liquid nitrogen and stored at −80°C. The other was fixed in 10% formalin for histopathological examination. The resected marginal normal tissues were frozen immediately in liquid nitrogen and stored at −80°C.

Microarray gene expression profiling

A total of 3 OSCC patients (2 tongue and 1 oral floor patient) were subjected to whole-genome analysis using microarray technology to determine the TSGs.

Total RNA was extracted from 3 OSCC patients using the Qiagen RNeasy mini kit and the SuperScript double-stranded cDNA synthesis kit (Invitrogen) was used to generate cDNA according to the manufacturer’s instructions. Cy3 labeling of ds-cDNA was performed overnight using the NimbleGen One-Color DNA labeling kit.

Cy3-labeled ds-cDNA (4 μg) was hybridized to the Homo sapiens 4×72 K gene expression array (Roche NimbleGen) representing 24,000 protein-coding genes, according to the manufacturer’s instructions. The mRNA expression data were analyzed using NimbleScan software version 2.4, which applied quintile normalization (18), and expression values were obtained using the Robust Multi-Chip Average algorithm as described by Irizarry et al (19). Expressional alterations of 2-fold across the two biological repeats were considered significant.

DNA copy number analysis by real-time QPCR

DNA from the frozen 60 tumor and marginal normal tissues was extracted using the QIAamp tissue kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions.

Real-time QPCR was performed using 60 tumor and marginal normal tissue samples with SYBR-Green I fluorescence detection on a LightCycler (Roche Diagnostics, Basel, Switzerland). Oligonucleotide primers for real-time QPCR were designed using Primer 3 software (Whitehead Institute for Biomedical Research), and uniqueness in the human genome was confirmed using a BLAST search. The primer set specific for CRISP3 was forward: 5′-ATCAGGCTGCATC CCAATAC-3′, and reverse: 5′-AACACCAAATCCCCACA GAA-3′. The 20-μl reaction mixture consisted of 10 μl 2X iQ SYBR-Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA), 10 ng genomic DNA, and 800 nM of each PCR primer. The reaction mixture was loaded into glass capillary tubes and submitted to an initial denaturation at 95°C for 10 min, followed by 45 cycles of amplification at 95°C for 10 sec for denaturation, 58°C for 10 sec for annealing, and 72°C for 15 sec for extension, with a temperature slope of 20°C/sec, performed in the LightCycler. The crossing point for each amplification curve was determined by the second derivative maximum method. The copy numbers are presented as the log ratio of each target locus in tumor normalization to internal reference loci (GAPDH) and relative to the normal DNA. The primer set specific to GAPDH was forward: 5′-CCACTAGGCGCTCACTGTTCT-3′, and reverse: 5′-GCG AACTCACCCGTTGACT-3′. DNA copy number loss was determined as <1.2 (20). The data were analyzed as the mean ± SD of three independent experiments with samples in triplicate.

Statistical analysis

The association between copy number loss and any clinical findings were assessed by the Fisher’s exact test. The association between the DNA copy number of tumor and normal tissues was assessed by the unpaired U test (SAS Institute, Cary, NC, USA). P-<0.05 was considered to indicate statistical significance.

Results

Clinicopathological findings

This study included 60 patients who had undergone surgical resection of primary OSCC. The patients, 45 males and 15 females, had average ages of 56.7 years for the males (range 31–85) and 63.1 years for the females (range 37–73). Regarding patient ethnicity,17 patients of Taiwanese and 43 of Japanese ethnicity were included in this study. The tumor sites were as follows: 23 patients, tongue; 13, buccal mucosa; 12, lower gingiva; 7, upper gingiva; and 5, oral floor. The T classifications, which indicate the sizes of the primary clinical tumors, were as follows: 15 patients with T1, 27 with T2, 10 with T3 and 8 with T4. The classifications by TNM stage were: 13 patients with Stage I, 12 with Stage II, 19 with Stage III, and 16 with Stage IV. A total of 18 of the 60 patients had histopathologically-confirmed cervical lymph node metastasis (LNM) at the time of diagnosis or during the 12-month follow-up period (LNM present).

Microarray analysis

Three OSCC patients were subjected to microarray analysis to screen for TSGs in this population. As in previous studies, we know that there are 17 upregulated genes in OSCC, comprising matrix metallopeptidase 1 MMP1, MMP10, MMP3, MMP12, PTHLH, INHBA, LAMC2, IL8, KRT17, COL1A2, IF16, ISG15, PLAU, GREM1, MMP9, IFI44 and CXCL1, and that there are 18 downregulated genes in OSCC, comprising KRT4, MAL, CRNN, SCEL, CRISP3, SPINK5, CLLA4, ADH1B, P11, TGM3, RHCG, PPP1R3C, CEACAM7, HPGD, CFD, ABCA8, CLU and CYP3A5 (15,21). In this study, we found 2 upregulated genes, MMP1 and MMP3, and 11 downregulated genes, CRISP3, SCGB3A1, AGR2, PIP, C20orf114, TFF1, STATH, AZGP1, MUC7, DMBT1 and LOC389429, in all three patients. The detected genes of up- and down-regulation are shown in Table II. CRISP3 was selected as it is currently unknown as to whether this gene is associated with OSCC.

Table II

List of aberrantly expressed genes.

Table II

List of aberrantly expressed genes.

Gene symbolGene nameLocation
Upregulated genes
MMP1Matrix metallopeptidase 111q22
MMP3Matrix metallopeptidase 311q22
Downregulated genes
STATHStatherin4q11-q13
MUC7Mucin 74q13-q21
SCGB3A1Secretoglobin, family 3A, member 15q35-qter
CRISP3Cysteine-rich secretory protein 36p12.3
LOC389429Hypothetical LOC3894296q21
AGR2Anterior gradient homolog 27p21.3
AZGP1α-2-glycoprotein 1, zinc-binding7q22.1
PIPProlactin-induced protein7q34
DMBT1Deleted in malignant brain tumors 110q26.13
C20orf114Chr. 20 open reading frame 11420q11.21
TFF1Trefoil factor 121q22.3
Assessment of CRISP3 by real-time QPCR

To evaluate microarray data at CRISP3, real-time QPCR was performed using non-amplified genomic DNA as a template. Initially, PCR primer sets located in CRISP3 were successfully designed to meet the criteria for reliable quantification. One PCR primer set was designed in a housekeeping gene, GAPDH, located on chromosome 12p, as a reference for normalization. Real-time QPCR analysis was performed with the above-mentioned primer sets using genomic DNA of 5 OSCC cell lines and 60 OSCC patients. DNA copy number loss of CRISP3 was observed in 2 of the 5 OSCC-derived cell lines (SAS and HSC2). The copy number of CRISP3 was significantly reduced in OSCC-derived cell lines compared with NOKs (P=0.024, Unpaired U-test: Fig. 1). The copy number loss of CRISP3 was observed in 24 (40%) of the 60 patient specimens. We evaluated statistically significant differences in all clinical characteristics pertaining to the CRISP3 copy number between OSCCs and normal tissues. No statistically significant differences were noted. Moreover, no statistically significant differences were observed in the CRISP3 copy number between OSCC tumor tissues and normal tissues in the 60 OSCC patients (P=0.401, Mann-Whitney’s U-test; Fig. 2).

Clinicopathological findings and statistical analysis

We compared our results with the clinicopathological findings for each tumor. The copy number loss of CRISP3 was observed in 24 (40%) of the 60 patients. The Fisher’s exact test was performed to evaluate the significance of correlations between copy number loss of CRISP3 and clinicopathological findings (Table III). A significant statistical correlation between the copy number loss and gender and T classification was observed. The copy number loss of CRISP3 was detected in early stage lesions; and the copy number loss of CRISP3 tended to be higher in early clinical stages. Moreover, no statistically significant correlation was found between the copy number loss of CRISP3 and other clinicopathological findings such as age, ethnicity, lymph node metastasis, tobacco and alcohol associated with the tumor samples.

Table III

Correlation between the DNA copy numbers of CRISP3 and clinical characteristics in OSCCs.

Table III

Correlation between the DNA copy numbers of CRISP3 and clinical characteristics in OSCCs.

LossNormalP-value
Gender
 Male13320.005
 Female114
Age
 <6517210.104
 ≥65157
Ethnic group
 Taiwanese6110.773
 Japanese1825
Tumor site
 Tongue6170.491
 Buccal mucosa67
 Lower gingival66
 Upper gingival43
 Oral floor23
T classification
 T1/T221210.021
 T3/T4315
LNM
 Present6120.573
 Absent1824
Stage
 I580.111
 II84
 III613
 IV511
Tobacco
 Yes7190.11
 No1717
Alcohol
 Yes12270.058
 No129

[i] LNM, lymph node metastasis confirmed by histopathological examination within the 12-month follow-up period.

Discussion

OSCC is often associated with loss of eating and speech function, disfigurement and psychological distress. The development of OSCC is strongly associated with smoking and excessive alcohol consumption (22). The prevention and management of this disease is likely to benefit from the identification of molecular markers and targets (7,8).

Recently, the development of tools for measuring gene expression and copy numbers across the entire genome has revolutionized our ability to characterize cancers at the molecular level. Cytogenetic analyses in conjunction with molecular genetics analyses by a number of research groups have shown an accumulation of genetic abnormalities during the development and/or progression of OSCC (23). In this study, we identified 11 downregulated genes, CRISP3, SCGB3A1, AGR2, PIP, C20orf114, TFF1, STATH, AZGP1, MUC7, DMBT1 and LOC389429, and 2 up-regulation genes, MMP1 and MMP3, using microarray technology. CRISP3 was selected due to the fact that it is unknown as to whether CRISP3 is associated with OSCC. However, to the best of our knowledge no previous OSCC studies have identified CRISP3 as we observed in the current study. Our findings indicate that DNA copy number loss of CRISP3 is involved in carcino-genesis in OSCC patients.

Little is known about the function of the mammalian CRISPs; however, CRISP1 and CRISP2 are known to be involved in various steps in reproduction (24). The C-terminal domain of CRISP2 has been shown to interact with calcium channels (25) and to bind a kinase present in the acrosome of mouse sperm (26). Less is known about human CRISP3. CRISP3, which is also known as specific granule protein of 28 kDa (SGP28), belongs to a family of CRISPs characterized by their size (220–230 amino acids), their secretory properties and a content of 16 highly conserved cysteine residues, which form an intra-molecular disulphide bond (27). Apart from its ability to bind A1BG in serum, Udby et al have shown that CRISP3 forms similar complexes with one of the three major proteins secreted from the prostate in seminal plasma, β-microseminoprotein (28). Human CRISP3 is also likely to be involved in the pathogenesis of prostate cancer, where CRISP3 expression is significantly upregulated (12). However, to the best of our knowledge no previous reports have identified the CRISP3 gene in OSCC.

Table II lists other candidate genes that may be associated with carcinogenesis in OSCC. These genes exhibit correlations with various types of carcinoma excluding C20orf114, STATH, and LOC389429. SCGB3A1 (HIN1) is a TSG that is highly expressed in a number of epithelial tissues, including the breast, lung, trachea, pancreas, prostrate and salivary gland. Inactivation of SCGB3A1 expression by promoter methylation is frequent in many types of epithelial carcinoma and carcinoma in situ, including breast, lung and nasopharyngeal carcinoma (29,30). AGR2 is a putative member of the protein disulfide isomerase family and was first identified as a homolog of the Xenopus laevis gene XAG-2. AGR2 was down-regulated in gastric tumor tissue compared to the control (31). AGR2 has previously been found to be one of several genes that encode secreted proteins showing an increased expression in prostate cancer cells compared to normal prostatic epithelium (32). We observed that the MMP1 gene is up-regulated in OSCC. MMP1 is located on chromosome 11q22.3 and belongs to the MMP family, which is responsible for the degradation of extracellular matrix components. There is clear evidence indicating that MMP1 is involved in various cell and tumour events, including cancer-cell development, growth, proliferation, apoptosis, invasion and metastasis, as well as angiogenesis and immune surveillance (3336).

Our results suggest that the CRISP3 gene is a novel TSG particular to OSCC, and inactivation of the CRISP3 gene may play one or more roles in the carcinogenesis of OSCCs. In the present study, we applied whole-genome analysis of TSGs in specimens from 3 patients with OSCC by microarray techno-logy. A significant statistical correlation was observed between the DNA copy number of CRISP3 and T-classification and gender. No significant correlation was found between DNA copy number loss of CRISP3 and age, ethnic group, tumor site, lymph node metastasis, tumor stage, tobacco or alcohol. In this population, 15 patients were habitual consumers of tobacco, alcohol and/or betel nuts. We compared these 15 patients with 22 patients who did not consume tobacco, alcohol or betel nuts. No significant correlation was found between the DNA copy number loss of CRISP3 and tobacco, alcohol and/or betel nut consumption. These results indicate that inactivation of the CRISP3 is an early event in OSCC, since T1/T2 classification is correlated with DNA copy number loss of CRISP3 rather than T3/T4 classification. We were not able to apply the functional analysis of the CRISP3 gene. Further studies employing techniques including immunoblotting, immunofluorescence, and immunohistochemistry are required to clarify the function of this gene in the development and progression of OSCC. Furthermore, a study with a larger patient series is requred to validate these results, in order that more appropriate treatment modalities can be offered to OSCC patients in Taiwan, Japan, and worldwide.

Acknowledgements

We thank Dr Homare Kawachi for his help with collecting OSCC samples and supplying the clinical data for the preparation of this manuscript.

References

1 

Sudbo J: Novel management of oral cancer: a paradigm of predictive oncology. Clin Med Res. 2:233–242. 2004. View Article : Google Scholar : PubMed/NCBI

2 

Macfarlane GJ, Zheng T, Marshall JR, Boyle P, et al: Alcohol, tobacco, diet and the risk of oral cancer: a pooled analysis of three case-control studies. Oral Oncol. 31:181–187. 1995. View Article : Google Scholar : PubMed/NCBI

3 

Mashberg A, Boffetta P, Winkelman R and Garfinkel L: Tobacco smoking, alcohol drinking, and cancer of the oral cavity and oropharynx among U.S. veterans. Cancer. 72:1369–1375. 1993. View Article : Google Scholar : PubMed/NCBI

4 

Warnakulasuriya S, Sutherland G and Scully C: Tobacco, oral cancer, and treatment of dependence. Oral Oncol. 41:244–260. 2005. View Article : Google Scholar

5 

Ries LAG, Harkins D, Krapcho M, et al: Cancer Statistics Review 1975–2003. National Cancer Institute; 2006

6 

Genden EM, Ferlito A, Bradley PJ, Rinaldo A and Scully C: Neck disease and distant metastases. Oral Oncol. 39:207–212. 2003. View Article : Google Scholar : PubMed/NCBI

7 

Sabichi AL, Demierre MF, Hawk ET, Lerman CE and Lippman SM: Frontiers in cancer prevention research. Cancer Res. 63:5649–5655. 2003.

8 

Spafford MF, Koch WM, Reed AL, et al: Detection of head and neck squamous cell carcinoma among exfoliated oral mucosal cells by microsatellite analysis. Clin Cancer Res. 7:607–612. 2001.PubMed/NCBI

9 

Fearon ER and Vogelstein B: A genetic model for colorectal tumorigenesis. Cell. 61:759–767. 1990. View Article : Google Scholar : PubMed/NCBI

10 

Marshall CJ: Tumor suppressor genes. Cell. 64:313–326. 1991. View Article : Google Scholar

11 

Garnis C, Baldwin C, Zhang L, Rosin MP and Lam WL: Use of complete coverage array comparative genomic hybridization to define copy number alterations on chromosome 3p in oral squamous cell carcinomas. Cancer Res. 63:8582–8585. 2003.PubMed/NCBI

12 

Bjartell A, Johansson R, Björk T, et al: Immunohistochemical detection of cysteine-rich secretory protein 3 in tissue and in serum from men with cancer or benign enlargement of the prostate gland. Prostate. 66:591–603. 2006. View Article : Google Scholar : PubMed/NCBI

13 

Horne AW, Duncan WC, King AE, et al: Endometrial cysteine-rich secretory protein 3 is inhibited by human chorionic gonadotrophin, and is increased in the decidua of tubal ectopic pregnancy. Mol Hum Reprod. 15:287–294. 2009. View Article : Google Scholar : PubMed/NCBI

14 

Udby L, Cowland JB, Johnsen AH, Sørensen OE, Borregaard N and Kjeldsen L: An ELISA for SGP28/CRISP-3, a cysteine-rich secretory protein in human neutrophils, plasma, and exocrine secretions. J Immunol Methods. 263:43–55. 2002. View Article : Google Scholar : PubMed/NCBI

15 

Ye H, Yu T, Temam S, et al: Transcriptomic dissection of tongue squamous cell carcinoma. BMC Genomics. 6:9–69. 2008.

16 

Kato H, Uzawa K, Onda T, et al: Down-regulation of 1D- myo-inositol 1,4,5-trisphosphate 3-kinase A protein expression in oral squamous cell carcinoma. Int J Oncol. 28:873–881. 2006.PubMed/NCBI

17 

UICC. TNM classification of malignant tumours. Fifth edition. pp. 20–24. 1997

18 

Bolstad BM, Irizarry RA, Astrand M and Speed TP: A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics. 19:185–193. 2003. View Article : Google Scholar : PubMed/NCBI

19 

Irizarry RA, Hobbs B, Collin F, et al: Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics. 4:249–264. 2003. View Article : Google Scholar

20 

Kuroiwa T, Yamamoto N, Onda T and Shibahara T: Expression of the FAM5C in tongue squamous cell carcinoma. Oncol Rep. 22:1005–1011. 2009.PubMed/NCBI

21 

Xu C, Liu Y, Wang P, et al: Integrative analysis of DNA copy number and gene expression in metastatic oral squamous cell carcinoma identifies genes associated with poor survival. Mol Cancer. 9:143–148. 2010. View Article : Google Scholar : PubMed/NCBI

22 

La Vecchia C: Epidemiology and prevention of oral cancer. Oral Oncol. 33:302–312. 1997.

23 

Martin CL, Reshmi SC, Ried T, et al: Chromosomal imbalances in oral squamous cell carcinoma: examination of 31 cell lines and review of the literature. Oral Oncol. 44:369–382. 2008. View Article : Google Scholar : PubMed/NCBI

24 

Cohen DJ, Busso D, Da Ros V, et al: Participation of cysteine-rich secretory proteins (CRISP) in mammalian sperm-egg interaction. Int J Dev Biol. 52:737–742. 2008. View Article : Google Scholar : PubMed/NCBI

25 

Gibbs GM, Scanlon MJ, Swarbrick J, et al: The cysteine-rich secretory protein domain of Tpx-1 is related to ion channel toxins and regulates ryanodine receptor Ca2+ signaling. J Biol Chem. 281:4156–4163. 2006. View Article : Google Scholar : PubMed/NCBI

26 

Gibbs GM, Bianco DM, Jamsai D, et al: Cysteine-rich secretory protein 2 binds to mitogenactivated protein kinase 11 in mouse sperm. Biol Reprod. 77:108–114. 2007. View Article : Google Scholar : PubMed/NCBI

27 

Gibbs GM and O’Bryan MK: Cysteine rich secretory proteins in reproduction and venom. Soc Reprod Fertil. 65:261–267. 2007.PubMed/NCBI

28 

Udby L, Lundwall Å, Johnsen AH, et al: β-microseminoprotein binds CRISP-3 in human seminal plasma. Biochem Biophys Res Commun. 333:555–561. 2005.

29 

Guo M, Ren J, Brock MV, Herman JG and Carraway HE: Promoter methylation of HIN-1 in the progression to esophageal squamous cancer. Epigenetics. 3:336–341. 2008. View Article : Google Scholar : PubMed/NCBI

30 

Castro M, Grau L, Puerta P, et al: Multiplexed methylation profiles of tumor suppressor genes and clinical outcome in lung cancer. J Transl Med. 8:86–96. 2010. View Article : Google Scholar : PubMed/NCBI

31 

Bai Z, Ye Y, Liang B, et al: Proteomics-based identification of a group of apoptosis-related proteins and biomarkers in gastric cancer. Int J Oncol. 38:375–383. 2011.PubMed/NCBI

32 

Maresh EL, Mah V, Alavi M, et al: Differential expression of anterior gradient gene AGR2 in prostate cancer. BMC Cancer. 13:680–687. 2010. View Article : Google Scholar : PubMed/NCBI

33 

Brinckerhoff CE and Matrisian LM: Matrix metalloproteinases: a tail of a frog that became a prince. Nat Rev Mol Cell Biol. 3:207–214. 2002. View Article : Google Scholar : PubMed/NCBI

34 

Egeblad M and Werb Z: New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2:161–174. 2002. View Article : Google Scholar : PubMed/NCBI

35 

Nagase H and Woessner JF Jr: Matrix metalloproteinases. J Biol Chem. 274:21491–21494. 1999. View Article : Google Scholar

36 

Ala-aho R and Kahari VM: Collagenases in cancer. Biochimie. 87:273–286. 2005. View Article : Google Scholar

Related Articles

Journal Cover

January 2012
Volume 3 Issue 1

Print ISSN: 1792-1074
Online ISSN:1792-1082

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Ko W, Sugahara K, Sakuma T, Yen C, Liu S, Liaw G and Shibahara T: Copy number changes of CRISP3 in oral squamous cell carcinoma. Oncol Lett 3: 75-81, 2012
APA
Ko, W., Sugahara, K., Sakuma, T., Yen, C., Liu, S., Liaw, G., & Shibahara, T. (2012). Copy number changes of CRISP3 in oral squamous cell carcinoma. Oncology Letters, 3, 75-81. https://doi.org/10.3892/ol.2011.418
MLA
Ko, W., Sugahara, K., Sakuma, T., Yen, C., Liu, S., Liaw, G., Shibahara, T."Copy number changes of CRISP3 in oral squamous cell carcinoma". Oncology Letters 3.1 (2012): 75-81.
Chicago
Ko, W., Sugahara, K., Sakuma, T., Yen, C., Liu, S., Liaw, G., Shibahara, T."Copy number changes of CRISP3 in oral squamous cell carcinoma". Oncology Letters 3, no. 1 (2012): 75-81. https://doi.org/10.3892/ol.2011.418