Secondary Logo

Journal Logo

Expression of programmed cell death protein 4 in nonmelanoma skin cancer compared with psoriasis and normal skin, a preliminary study

Abdel-Halim, Mona R.E.; Mohammed, Faisal N.; Sayed, Khadiga S.; Abouelfadl, Dalia M.; Gamal El-Din, Amina A.; Badawi, Manal A.

Journal of the Egyptian Women's Dermatologic Society: May 2016 - Volume 13 - Issue 2 - p 77–82
doi: 10.1097/01.EWX.0000481053.61341.ae
Original articles
Free

Background Tumors expressing programmed cell death protein 4 (PDCD4) are associated with a better prognosis. Limited work has been carried out on PDCD4 expression in nonmelanoma skin cancer.

Objective To study the expression of PDCD4 in basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) in comparison with psoriasis and normal skin.

Patients and methods Using immunohistochemistry, the expression of PDCD4 was studied in nine BCC, five SCC, ten psoriasis, and six normal skin biopsies.

Results All BCC specimens showed negative PDCD4 expression, with focal nuclear positivity detected only in three cases. Well-differentiated areas of SCC showed positive (mainly cytoplasmic) staining of PDCD4, whereas undifferentiated areas were negative. In psoriasis, suprabasal PDCD4 cytoplasmic positivity was detected in eight cases. All normal skin biopsies showed suprabasal nuclear staining of PDCD4.

Limitation Small number of cases in each group.

Conclusion This study preliminarily supports a potential role of PDCD4 in the suppression of nonmelanoma skin cancer.

aDepartment of Dermatology Department, Dermatopathology Unit

bDepartment of Dermatology, Faculty of Medicine, Cairo University, Giza

Departments of cDermatology

dPathology, National Research Centre, Cairo, Egypt

Correspondence to Mona R.E. Abdel-Halim, MD, 108 Nile Street, Dokki, Giza, 12311 Cairo, Egypt Tel: +20 238 246 595; fax: +20 237 494 428; e-mail: abdelhalimmona@gmail.com

Received June 5, 2015

Accepted January 27, 2016

Back to Top | Article Outline

Introduction

The term nonmelanoma skin cancer (NMSC) includes squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) 1. Many factors contribute toward the increased incidence of NMSC, and these are mainly linked to activation of proto-oncogenes and inhibition of tumor-suppressor genes 2.

Programmed cell death protein 4 (PDCD4) is a tumor-suppressor gene that was identified to be upregulated in apoptosis 3,4. It influences transcription, translation, and signal transduction pathways in a way that suppresses tumorous growth 5. Experimental studies showed that PDCD4 could suppress the transformation of mouse epidermal cells exposed to different carcinogenesis promoters 6,7. Accordingly, its high expression in tumors was associated with less aggressiveness and a better prognosis 8.

As limited work has been carried out on the expression of PDCD4 in NMSC, the current study aimed to study its expression in BCC and SCC in comparison with psoriasis (benign proliferative skin disease) and normal skin.

Back to Top | Article Outline

Patients and methods

The study was approved by the Dermatology Department Research Ethical Committee. Deparaffinized sections of 3 µm cut from routinely fixed and paraffin-embedded specimens of BCC (nine cases), SCC (five cases, moderately differentiated), psoriasis vulgaris (10 cases), and normal skin (six biopsies) were stained immunohistochemically with rabbit antihuman PDCD4 monoclonal antibody (EPR3431) (Novus Biologicals, Littleton, Colorado, USA) using the avidin–biotin complex peroxidase staining technique 9. The intensity and pattern of staining were assessed. In addition, a morphometric analysis was carried out using the Leica Qwin 500 Image Analyzer (LEICA Imaging Systems Ltd, Cambridge, UK), which consists of a Leica DM-LB microscope with a JVC color video camera attached to a computer system. Marker color was first detected, followed by the formation of a binary image for the area stained by the marker. The mean area percent of PDCD4 staining was estimated for each studied case.

Photomicrographs depicting the various findings were obtained using a Canon Power Shot G15 digital camera (Canon, New York, New York, USA), connected to a Primo Star Zeiss Microscope (Zeiss, Oberkochen, Germany). All photomicrographs presented are according to their original magnification.

Back to Top | Article Outline

Results

Apart from strong focal nuclear positivity in the basaloid cells in three (33.3%) BCC cases (Fig. 1), all BCC cases showed negative PDCD4 expression in the tumor islands (Fig. 2). Well-differentiated areas of SCC showed moderately positive (mainly cytoplasmic) staining of PDCD4 (Fig. 3), whereas undifferentiated areas were negative (Fig. 4). In psoriasis, moderate predominantly suprabasal PDCD4 cytoplasmic positivity was detected in eight (80%) cases (Fig. 5). This was diffuse in three cases, patchy in three cases, and focal in two cases. Negative expression was detected in two (20%) psoriatic cases. All normal skin biopsies showed epidermal nuclear staining of PDCD4 mainly in suprabasal layers (Fig. 6). Staining was also strongly detected in sebaceous lobules, ductal, and secretory portions of sweat glands. In all specimens, staining of inflammatory cells, fibroblasts, sebaceous lobules, and sweat gland structures was considered a positive internal control.

Figure 1

Figure 1

Figure 2

Figure 2

Figure 3

Figure 3

Figure 4

Figure 4

Figure 5

Figure 5

Figure 6

Figure 6

Table 1 shows the results of morphometric analysis of PDCD4 staining in all studied specimens. No significant difference was found in PDCD4 staining in BCC, SCC, and psoriasis compared with normal skin (P=0.1, 0.4, and 0.9, respectively). Also, no significant difference was found on comparing BCC and SCC with psoriasis (P=0.25 and 0.4, respectively).

Table 1

Table 1

Back to Top | Article Outline

Discussion

In agreement with Matsuhashi et al.10, almost no PDCD4 expression was detected in BCC cases. Focal nuclear positivity was detected only in three cases. Interestingly, this focal nuclear positivity was detected in areas of the tumor showing scattered apoptotic bodies on hematoxylin and eosin examination, providing more evidence of the role of PDCD4 in promoting apoptosis of epithelial cells.

To date, PDCD4 expression in cutaneous SCC has not been studied previously. However, its expression has been studied in laryngeal SCC 11, esophageal SCC 12, oral SCC 13, and in lung cancer including SCC 8. Similar to our findings, these studies reported loss of PDCD4 expression in high-grade undifferentiated areas (cases) of SCC. Because PDCD4 contributes toward the differentiation of keratinocytes by inhibiting AP-1 activities 7,14 and activating caspase-mediated pathways 10,15, well-differentiated areas of SCC showed evident PDCD4 expression.

Suprabasal cytoplasmic expression of PDCD4 was detected in 80% of psoriasis cases. It is speculated that this cytoplasmic upregulation of PDCD4 in psoriatic skin represents an additional mechanism, together with P16-induced senescence of keratinocytes 16, in preventing neoplastic transformation in this highly proliferative skin disorder.

Suprabasal nuclear expression of PDCD4 in all normal skin biopsies in the current study supports previous results that reported its expression in differentiating (suprabasal) layers of the skin and hair follicles (suprabulbar) and its absence or reduced expression in the proliferating basal cell layer and in the hair bulb 10.

Although normal skin showed nuclear staining of PDCD4 in differentiating suprabasal layers, well-differentiated areas of SCC and psoriatic epidermis showed cytoplasmic staining. In most studies on cancer cell lines, cytoplasmic expression of PDCD4 was detected 17. This may reflect a reaction of the keratinocytes to changes in its microenvironment both in psoriasis and in SCC, with subsequent increased activity of differentiating pathways and altered expression patterns.

Back to Top | Article Outline

Conclusion

This study provides preliminary support for a potential role of PDCD4 in tumor suppression in NMSC. Therapeutic approaches that ultimately lead to upregulation of PDCD4 expression in tumor cells with subsequent promotion of tumor cell apoptosis may represent a new approach in NMSC therapy. Further studies on larger number of cases are still needed to validate these results.

Back to Top | Article Outline

Acknowledgements

The authors would like to thank all residents in the Dermatology and Surgery Departments at Cairo University and National Research Centre for helping us with the recruitment of cases.

Back to Top | Article Outline

Conflicts of interest

There are no conflicts of interest.

Back to Top | Article Outline

References

1. Diepgen TL, Mahler V. The epidemiology of skin cancer. Br J Dermatol 2002; 146 (Suppl 61):1–6.
2. Stratigos AJ, Kapranos N, Petrakou E, Anastasiadou A, Pagouni A, Christofidou E, et al.. Immunophenotypic analysis of the p53 gene in non-melanoma skin cancer and correlation with apoptosis and cell proliferation. J Eur Acad Dermatol Venereol 2005; 19:180–186.
3. Matsuhasi S, Yoshinaga H, Yatsuki H, Tsugita A, Hori K. Isolation of a novel gene from a human cell line with Pr-28 MAb which recognizes a nuclear antigen involved in the cell cycle. Res Commun Biochem Cell Mol Biol 1997; 1:109–120.
4. Young MR, Yang HS, Colburn NH. Promising molecular targets for cancer prevention: AP-1, NF-kappa B and Pdcd4. Trends Mol Med 2003; 9:36–41.
5. Shibahara K, Asano M, Ishida Y, Aoki T, Koike T, Honjo T. Isolation of a novel mouse gene MA-3 that is induced upon programmed cell death. Gene 1995; 166:297–301.
6. Cmarik JL, Min H, Hegamyer G, Zhan S, Kulesz-Martin M, Yoshinaga H, et al.. Differentially expressed protein Pdcd4 inhibits tumor promoter-induced neoplastic transformation. Proc Natl Acad Sci USA 1999; 96:14037–14042.
7. Yang HS, Jansen AP, Nair R, Shibahara K, Verma AK, Cmarik JL, Colburn NH. A novel transformation suppressor, Pdcd4, inhibits AP-1 transactivation but not NF-kappaB or ODC transactivation. Oncogene 2001; 20:669–676.
8. Chen Y, Knösel T, Kristiansen G, Pietas A, Garber ME, Matsuhashi S, et al.. Loss of PDCD4 expression in human lung cancer correlates with tumour progression and prognosis. J Pathol 2003; 200:640–646.
9. Cote A, da Silva R, Cuello A. Cuello A. Current protocols for light microscopcy immunocytochemistry. Immunohistochemistry II. Chichester, UK: John Wiley and Sons; 1993. 147–168.
10. Matsuhashi S, Narisawa Y, Ozaki I, Mizuta T. Expression patterns of programmed cell death 4 protein in normal human skin and some representative skin lesions. Exp Dermatol 2007; 16:179–184.
11. Wang J, Zhang Y. Expression of programmed cell death 4 and its correlation with proliferation and apoptosis in laryngeal squamous cell carcinoma. Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi 2012; 26:266–269.
12. Fassan M, Realdon S, Pizzi M, Balistreri M, Battaglia G, Zaninotto G, et al.. Programmed cell death 4 nuclear loss and miR-21 or activated Akt overexpression in esophageal squamous cell carcinogenesis. Dis Esophagus 2012; 25:263–268.
13. Reis PP, Tomenson M, Cervigne NK, Machado J, Jurisica I, Pintilie M, et al.. Programmed cell death 4 loss increases tumor cell invasion and is regulated by miR-21 in oral squamous cell carcinoma. Mol Cancer 2010; 9:238.
14. Yang HS, Knies JL, Stark C, Colburn NH. Pdcd4 suppresses tumor phenotype in JB6 cells by inhibiting AP-1 transactivation. Oncogene 2003; 22:3712–3720.
15. Zhang H, Ozaki I, Mizuta T, Hamajima H, Yasutake T, Eguchi Y, et al.. Involvement of programmed cell death 4 in transforming growth factor-beta1-induced apoptosis in human hepatocellular carcinoma. Oncogene 2006; 25:6101–6112.
16. Nickoloff BJ. Creation of psoriatic plaques: the ultimate tumor suppressor pathway. A new model for an ancient T-cell-mediated skin disease. Viewpoint. J Cutan Pathol 2001; 28:57–64.
17. Yoshinaga H, Matsuhashi S, Fujiyama C, Masaki Z. Novel human PDCD4 (H731) gene expressed in proliferative cells is expressed in the small duct epithelial cells of the breast as revealed by an anti-H731 antibody. Pathol Int 1999; 49:1067–1077.
Keywords:

basal cell carcinoma; nonmelanoma skin cancer; programmed cell death protein 4; psoriasis; squamous cell carcinoma

© 2016 Egyptian Women's Dermatologic Society