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ORIGINAL ARTICLE
Year : 2011  |  Volume : 7  |  Issue : 2  |  Page : 180-182

Serum phosphodiesterase levels in oral cancer


1 Department of Biochemistry, Kasturba Medical College, Manipal University, Manipal, India
2 Department of Surgical Oncology, Kasturba Medical College, Manipal University, Manipal, India
3 Department of Radiotherapy, Kasturba Medical College, Manipal University, Manipal, India
4 Department of Community Medicine, Kasturba Medical College, Manipal University, Manipal, India

Date of Web Publication12-Jul-2011

Correspondence Address:
K Prabhu
Department of Biochemistry, Kasturba Medical College, Manipal University, Manipal
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0973-1482.82911

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 > Abstract 

Background: Oral squamous cell carcinoma (OSCC) is one of the most common malignancies recognized nowadays. Its early detection is the better alternative to provide a good quality of life for the patients. During the last years, several studies have identified potential biomarkers of OSCC progression and prognosis. The phosphodiesterases (PDEs) are responsible for the hydrolysis of the second messengers with a fundamental role in the transduction of the intracellular signals. Variations in PDE activity have been correlated to different pathological mechanisms, such as cellular differentiation, apoptosis, and tumor invasivity. PDEs are also known to play a role in tumor growth by influencing angiogenesis.
Aim: To estimate and compare serum PDE levels in healthy controls and biopsy-proven oral cancer patients before definitive therapy.
Materials and Methods: Institutional Ethics Committee gave us the permission to conduct this study. After obtaining consent from biopsy-proven oral cancer patients (n = 39) (before onset of any definitive treatment) and age- and sex-matched healthy controls (n = 20), 2 ml of blood was collected in plain vacutainers. After clot formation, samples were centrifuged and serum was collected for estimation of PDE.
Statistical Analysis: Kruskal-Wallis test; Mann-Whitney Test
Results and Discussion: Pretreatment PDE levels were significantly elevated in oral cancer patients (P<0.0001) as compared with the controls and also there was a significant increase in PDE levels (P<0.001) with advancing stage in oral cancer patients. This may implicate a role for serum PDE in pathophysiology of oral cancer.

Keywords: Oral cancer, pathophysiology, phosphodiesterase


How to cite this article:
Prabhu K, Naik D, Ray S, Vadiraja B M, Kamath A. Serum phosphodiesterase levels in oral cancer. J Can Res Ther 2011;7:180-2

How to cite this URL:
Prabhu K, Naik D, Ray S, Vadiraja B M, Kamath A. Serum phosphodiesterase levels in oral cancer. J Can Res Ther [serial online] 2011 [cited 2019 Sep 20];7:180-2. Available from: http://www.cancerjournal.net/text.asp?2011/7/2/180/82911


 > Introduction Top


Cancer affects all communities worldwide and approximately 10 million people are diagnosed with cancer. Cancer is a class of disease in which a group of cells display uncontrolled growth, invasion, and angiogenesis. Tumor angiogenesis or neovascularization is the proliferation of a network of blood vessels that penetrate into cancerous growths, supplying nutrients and oxygen and removing waste products. [1]

Oral cancer is the eighth most common cancer worldwide, [2] prevalence being high among men. In India, the age-standardized incidence rate of oral cancer is 12.6 per 1 00 000 population. [3] "Oral" cancer per se includes cancers of the lip, tongue, gingiva, oral mucosa, oropharynx, and hypopharynx. [4] The clinical and histological features alone cannot always accurately predict whether potentially malignant disorders of the oral mucosa remain stable, regress, or progress to malignancy. [5] Identification of molecular markers (or biomarkers) which can predict disease progression is necessary for better management of these disorders. [5] Commonly, nonfunctional plasma enzymes are used as an aid for diagnosis and prognosis, e.g., aminotransferases, amylase, [6] etc. Markers like glutathione S-transferases, N-acetyltransferases, and serum dipeptidyl peptidase have been associated with various epithelial malignancies and have been shown to influence the susceptibility for cancer and outcome of treatment. [7],[8],[9],[10],[11] Alteration in PDE activity has been associated with cellular differentiation, apoptosis, tumor invasion, [Figure 1]. [12],[13] angiogenesis, etc. [14]
Figure 1: Figure showing mechanism of PDE in carcinogenesis

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Aim

To estimate and compare serum phosphodiesterase (PDE) levels in healthy controls and biopsy-proven oral cancer patients before definitive therapy.


 > Materials and Methods Top


The study was carried out after obtaining approval from the Institutional Ethics Committee. In this study, 59 subjects, aged between 25 and 75 years of either sex, were used. Of them, 39 were biopsy-proven oral cancer patients (stage 2, 3, and 4) and 20 were healthy controls. The oral cancer patients were admitted under Radiotherapy and Oncology department between June 2009 and June 2010. Cancer cases with associated serious diseases like liver disease, diabetes, renal disease, and those who were on any long-term medications were excluded. Cases were selected at our convenience. They received radiotherapy as definitive treatment. The control group comprised of age- and gender-matched healthy subjects. Among oral cancer patients, 31 were males and 8 were females.

Blood (2 ml) was collected from controls after obtaining consent. The blood samples (2 ml) were also collected with prior consent from cancer patients before radiotherapy. The blood samples were collected in a vacutainer with no anticoagulant. The sample was allowed to clot for 30 minutes. After centrifugation at 3 000 rpm for five minute, the clear supernatant serum was used for the determination of PDE levels. PDE levels of samples were measured by using Genesis 10 UV (Thermo Electron Corporation). For maintaining the reaction temperature at 37°C, automated water bath obtained from Rotek was used. Cyber scan 510 pH meter (Elico Ltd) was used. Sartorius balance was used for weighing chemicals.

PDE assay

Principle

Paranitrophenyl phosphate (4-nitrophenyl phosphate) is hydrolyzed by PDE to 4-nitrophenol (4-hydroxynitrobenzene) and inorganic phosphate. The yellow color formed due to liberation of 4-nitrophenolate at pH 9 was measured spectrophotometrically at 400 nm. [15]

Procedure

In a clean dry test tube, 1 ml of assay mixture containing 500 μl of Tris HCl, 100 μl of MgCl 2, 100 μl of paranitrophenyl phosphate, and 300 μl of distilled water was taken. The mixture was incubated at 37°C for 5 minutes. Then, to this mixture, 10 μl of serum (10 μl of double distilled water in case of blank) was added. The mixture was again incubated at 37°C for 10 minutes. 2 ml of NaOH with EDTA was added to each test tube to stop the reaction. Activity was calculated using molar absorption coefficient of the product of chemical reaction, 4-nitrophenol (17, 600). [15]


 > Results Top


The patients (n = 39) of oral cancer presented with either stage 2 or 3 or 4 of cancer. In the group comprising of oral cancer patients, pretreatment PDE levels were significantly elevated (P≤0.0001) as compared with the controls [Table 1]. Pretreatment serum PDE levels showed an increase with advancing stages of oral cancer [Table 2]. On stage-wise comparison of pretreatment serum PDE levels using Kruskal Wallis test, a significant (P<0.001) difference was observed. So, a pair-wise comparison between stages was made--there was a significant (P<0.001) difference in PDE levels of stage 4 as compared with that of stage 2 and stage 3 [Table 2].
Table 1: Comparison of pretreatment serum PDE levels of oral cancer patients with healthy controls

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Table 2: Comparison of pretreatment serum PDE levels in between stages of oral cancer patients

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 > Discussion Top


Oral squamous cell carcinoma (OSCC) is one of the most common malignancies recognized nowadays, and represents a public health problem. Its early detection helps to provide a good quality of life for the patients. During the past years, several studies have identified potential biomarkers of OSCC progression and prognosis. [16]

cAMP is an important "second messenger" transferring information into cells and exists ubiquitously in the tissues and cells of mammals. The cAMP signal pathway is involved in many metabolic pathways in cells, and regulates many physiological processes, such as cell metabolism, proliferation, and cell death. The PDEs are responsible for the hydrolysis of the second messengers, with a fundamental role in the transduction of the intracellular signals. Variations in PDE activity have been found in different pathologies, and they have also been correlated to different pathophysiological mechanisms, such as cellular differentiation, apoptosis, and tumor invasivity. [13],[17] PDEs are also known to play a role in tumor growth by influencing angiogenesis. [18],[19] Inhibition of selective PDE isoforms, which raises the levels of intracellular cAMP and cGMP, has been shown to induce apoptosis and cell cycle arrest in a broad spectrum of tumor cells. [20],[21],[22] Our results showed a significant increase in pretreatment PDE levels in cancer patients as compared with controls. In our study, pretreatment serum PDE levels correlated well with advancing stage of cancer and also showed higher levels in cancer patients than controls. This implies that they might have been induced and released from cancer cells. This may imply a possible role for inhibitors of specific PDE isoenzymes which may selectively restore normal intracellular signaling, providing antitumor therapy with reduced adverse effects. [19]

A limitation of the present study is lesser number of controls as compared with cases. The disparity in the number was due to lack of healthy volunteers consenting for the study.

 
 > References Top

1.Gupta MK, Qin RY. Mechanism and its regulation of tumor-induced angiogenesis. World J Gastroenterol 2003;9:1144-55.  Back to cited text no. 1
    
2.Petersen PE. Strengthening the prevention of oral cancer: The WHO perspective. Community Dent Oral Epidemiol 2005;33:397-9.  Back to cited text no. 2
    
3.Park K. Cancer. In: Park K, editor. Textbook of Preventive and Social Medicine. 20 th ed. Jabalpur: Banarsidas Bhanot Publishers; 2009. p. 332-40.  Back to cited text no. 3
    
4.Miller BA. Cancer/gov [homepage on the net]. Racial/Ethnic Patterns of Cancer in the United States. 1988:49. Available from: http://seer.cancer.gov/publications/ethnicity/oralcav.pdf. [Last accessed on 2010 Sep 27].  Back to cited text no. 4
    
5.Pitiyage G, Tilakaratne WM, Tavassoli M, Warnakulasuriya S. Molecular markers in oral epithelial dysplasia: Review. J Oral Pathol Med 2009;38:737-52.  Back to cited text no. 5
    
6.Murray RK, Granner DK, Rodwell VW, Kennelly PJ. Enzymes. In: Kennelly PJ, editor. Harper′s Illustrated Biochemistry. 26 th ed. New Delhi: Mc Graw Hill Lange; 2008. p. 57-9.  Back to cited text no. 6
    
7.Hengstler JG, Arand M, Herrero ME, Oesch F. Polymorphisms of N-acetyltransferases, glutathione S-transferases, microsomal epoxide hydrolase and sulfotransferases: Influence on cancer susceptibility. Recent Results Cancer Res 1998;154:47-85.  Back to cited text no. 7
    
8.Urade M, Uematsu T, Mima T, Ogura T, Matsuya T. Serum dipeptidyl peptidase (DPP) IV activity in hamster buccal pouch carcinogenesis with 9, 10-dimethyl-1, 2-benzanthracene. J Oral Pathol Med 1992;21:109-12.  Back to cited text no. 8
    
9.Prabhu K, Bhat GP. Serum total glutathione-S-transferase levels in oral cancer. J Cancer Res Ther 2007;3:167-8.  Back to cited text no. 9
    
10.Ferrandina G, Scambia G, Damia G, Tagliabue G, Fagotti A, Benedetti Panici P, et al. Glutathione S-transferase activity in epithelial ovarian cancer: association with response to chemotherapy and disease outcome. Ann Oncol 1997;8:343-50.  Back to cited text no. 10
    
11.Singh M, Shah PP, Singh AP, Ruwali M, Mathur N, Pant MC, et al. Association of genetic polymorphisms in glutathione S-transferases and susceptibility to head and neck cancer. Mutat Res 2008;638:184-94.  Back to cited text no. 11
    
12.Spoto G, Fioroni M, Rubini C, Di Nicola M, Di Pietrantonio F, Di Matteo E, et al. Cyclic AMP phosphodiesterase activity in human gingival carcinoma. J Oral Pathol Med 2004;33:269-73.  Back to cited text no. 12
    
13.Prosperi C, Scali C, Barba M, Bellucci A, Giovannini MG, Pepeu G, et al. Comparison between flurbiprofen and its nitric oxide-releasing derivatives HCT-1026 and NCX-2216 on Abeta(1-42)-induced brain inflammation and neuronal damage in the rat. Int J Immunopathol Pharmacol 2004;17:317-30.  Back to cited text no. 13
    
14.Ptaszynska MM, Pendrak ML, Bandle RW, Stracke ML, Roberts DD. Positive feedback between vascular endothelial growth factor-A and autotaxin in ovarian cancer cells. Mol Cancer Res 2008;6:352-63.  Back to cited text no. 14
    
15.Grossman L, Moldave K. Purification and properties of Venom Phosphodiesterase. In: Laskowski M, editor. Methods in enzymology, Nucleic acids. Part I, vol 65. New York: Academic press; 1974. p. 279-80.  Back to cited text no. 15
    
16.Hoffmann RR, Yurgel LS, Campos MM. Endothelins and their receptors as biological markers for oral cancer. Oral Oncol 2010;46:644-7.   Back to cited text no. 16
    
17.Spoto G, della Malva M, Rubini C, Fioroni M, Piattelli A, Serra E, et al. cAMP phosphodiesterase activity evaluation in human carcinoma of salivary glands. Nucleosides Nucleotides Nucleic Acids 2006;25:1113-7.  Back to cited text no. 17
    
18.Chougule A, Hussain S, Agarwal DP. Prognostic and diagnostic value of serum pseudocholinesterase, serum aspartate transaminase, and serum alinine transaminase in malignancies treated by radiotherapy. J Cancer Res Ther 2008;4:21-5.  Back to cited text no. 18
    
19.Savai R, Pullamsetti SS, Banat GA, Weissmann N, Ghofrani HA, Grimminger F, et al. Targeting cancer with phosphodiesterase inhibitors. Expert Opin Investig Drugs 2010;19:117-31.  Back to cited text no. 19
    
20.Dou AX, Wang X. Cyclic adenosine monophosphate signal pathway in targeted therapy of lymphoma. Chin Med J (Engl) 2010;123:95-9.  Back to cited text no. 20
    
21.Son J, Lee JH, Kim HN, Ha H, Lee ZH. CAMP-response-element-binding protein positively regulates breast cancer metastasis and subsequent bone destruction. Biochem Biophys Res Commun 2010;398:309-14.  Back to cited text no. 21
    
22.Safa M, Zand H, Mousavizadeh K, Kazemi A, Bakhshayesh M, Hayat P. Elevation of cyclic AMP causes an imbalance between NF-kappaB and p53 in NALM-6 cells treated by doxorubicin. FEBS Lett 2010;584:3492-8.  Back to cited text no. 22
    


    Figures

  [Figure 1]
 
 
    Tables

  [Table 1], [Table 2]



 

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