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ORIGINAL ARTICLE
Year : 2021  |  Volume : 34  |  Issue : 3  |  Page : 941-946

Immunohistochemical expression of myxovirus resistant protein 1 in squamous cell carcinoma


1 Dermatology, Andrology and STDs Department, Faculty of Medicine, Menoufia University, Egypt
2 Dermatology, Andrology and STDs Department, Berket El-Sabae General Hospital, Berket El-Sabae, Menoufia, Egypt

Date of Submission03-May-2020
Date of Decision15-Jun-2020
Date of Acceptance19-Jun-2020
Date of Web Publication18-Oct-2021

Correspondence Address:
Walaa W Fayed
Berket El-Sabae, Menoufia
Egypt
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/mmj.mmj_148_20

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  Abstract 


Objective
The aim was to evaluate the role of myxovirus resistant protein 1 (MxA) in cutaneous squamous cell carcinoma (CSCC).
Background
CSCC is the second most common skin cancer. MxA has the potential importance in tumorigenesis and metastasis as well as in the treatment and prognosis of different cancers such as CSCC. However, the clear association between MxA expression and cancer remains unknown.
Patients and methods
This prospective and retrospective case-controlled study was conducted on 48 cases with CSCC and 43 age- and sex-matched apparently healthy participants. All sections were immunohistochemically stained for MxA antibody.
Results
Significant differences between control and CSCC groups regarding MxA positivity in both tumor and stroma were identified. All CSCC cases were positive in tumor and stroma, whereas 44.2% of control cases showed positivity (P < 0.001 for both). Overexpression of MxA in tumor was significantly associated with high grade and advanced stage (P = 0.02 and 0.03, respectively). Overexpression of MxA in stroma was significantly associated with low grade and early stage (P = 0.02 and 0.02, respectively).
Conclusion
The role of MxA is complex and controversial in tumor and stroma of CSCC. Tumor MxA expression is responsible for tumor progression, whereas stromal expression is responsible for regression.

Keywords: cutaneous squamous cell carcinoma, immunohistochemistry, marker, myxovirus resistant protein 1


How to cite this article:
Samaka RM, Basha MA, Fayed WW. Immunohistochemical expression of myxovirus resistant protein 1 in squamous cell carcinoma. Menoufia Med J 2021;34:941-6

How to cite this URL:
Samaka RM, Basha MA, Fayed WW. Immunohistochemical expression of myxovirus resistant protein 1 in squamous cell carcinoma. Menoufia Med J [serial online] 2021 [cited 2024 Mar 29];34:941-6. Available from: http://www.mmj.eg.net/text.asp?2021/34/3/941/328299




  Introduction Top


Cutaneous squamous cell carcinoma (CSCC) is a malignancy of epidermal keratinocytes that exhibits various degrees of differentiation [1]. It is the second most common cutaneous malignancy [2].

The pathogenesis of CSCC is usually combined with environmental factors, mainly ultraviolet irradiation through long-term sun exposure. Ultraviolet light can randomly induce DNA damage in keratinocytes. Moreover, it can mutate genes essential for control and surveillance in the skin epidermi., CSCC most often develops on areas of skin regularly exposed to the sun, such as the face, ears, hands, shoulders, upper chest, and back [3].

The myxovirus resistance 1 (M × 1) gene is one of the most prominent interferon (IFN)-stimulated genes in vertebrate that are highly activated when triggered by type I and III IFNs upon viral infection. Myxovirus resistant protein 1 (MxA) is thought to be a useful biomarker for monitoring IFN activity and predicting clinical efficacy during IFN therapy in patients with certain types of cancer [4]. MxA is associated with different types of human cancer. Although MxA plays a role in antitumor activity, it remains to be unclear how MxA affects tumor-infiltrating immune cells in cancer [5]. The role of MxA in tumor is still controversial, especially in CSCC, so our study aimed to evaluate the role of MxA in CSCC.


  Patients and methods Top


This retrospective and prospective case-controlled study included 48 cases with CSCC and 43 age-matched and sex-matched healthy participants as a control group. This study was approved by the Ethical Committee in Menoufia University. Written informed consent was obtained from participants. Newly diagnosed CSCC cases were selected from Dermatology Outpatient Clinic and plastic Surgery Department, Menoufia University Hospital, during the period from January 2016 to December 2017. For CSCC cases, they were selected, based on the availability of paraffin-embedded blocks for serial cutting and examination. Tissue blocks were retrieved from archives of Pathology Department, Faculty of Medicine, Menoufia University Hospital. Prospective collected cases surgically biopsied were fixed in 10% formalin solution, and then sent to Pathology Department, Faculty of Medicine, Menoufia University, where they were subjected to routine tissue processing and preparation of hematoxylin and eosin-stained slides for assessment of pathological changes. Normal skin samples were obtained from persons attending Plastic Surgery Department.

Tissue microarray was done on more representative parts of the section

Immunohistochemistry: immunohistochemical staining was performed using rabbit anti-MxA polyclonal antibody (Cat. # GTX110256; isotype IgG; diluted at 1: 100; Gene Tex, California, 2456 Alton Pkwy Irvine, CA 92606 USA). All slides were de-paraffinzed using xylene and then rehydrated in decreased concentrations of ethanol. Antigen retrieval was done by using microwave heating (20 min; 10 mmol/citrate buffer, pH 6. 0) after inhibition of endogenous peroxidase activity (hydrogen peroxidase for 15 min).

The primary antibodies were applied on the slides and incubated overnight at room temperature in a humid chamber. Finally, the detection of bound antibody was done by using a modified labeled avidin–biotin (LAB) reagent for 20 min and then PBS wash. We used a 0. 1% solution of diaminobenzidine for 5 min as a chromogen. Slides were counterstained with Mayer's hematoxylin for 5–10 min. Negative control slides were prepared, by removing the primary antibodies from the staining procedure. Human colon carcinoma was used as a positive control for MxA.

Statistical analysis

The statistical analysis was conducted using Statistical Package for the Social Sciences program for Windows (version 20; SPSS Inc., Chicago, Illinois, USA). Qualitative data are expressed as numbers and percentages. Quantitative data are expressed as arithmetic mean, SD, percentage, and median. χ2 test and Fisher's exacts test were used to compare qualitative variables. Mann–Whitney U-test was used in comparing two nonparametric quantitative variables. Kruskal–Wallis test was used to test the equality of population medians between groups, as it is considered an extension of the Mann–Whitney test. Spearman correlation was also used. The results were statistically significant for P value less than 0.05.


  Results Top


Clinicopathological data of the studied CSCC group (data not shown in tables) revealed the following: patients with CSCC had age ranged from 6 to 78 years, with a mean ± SD of 51. 96 ± 19. 59 years. They were equally distributed between males and females (50% for each), with male to female ratio of 1: 1. Extremities were affected in six (12.5%) cases, whereas head and neck were affected in 30 (62.5%) cases, and trunk was affected in 12 (25%) cases. The studied group clinically presented as ulcer in 14 (29.2%) cases and as mass in 34 (70.8%) cases. The margin was involved in four (8.3%) cases, cannot be assessed in 12 (25%) cases, and not involved in 32 (66.7%) cases. The size of the tumor ranged between 0.5 and 15 cm, with 3.88 ± 3.26 as a mean ± SD value and 3 as a median. Lymphovascular invasion and perineural invasion were absent in all 43 (100%) cases. Grade grouping in included cases was identified as grade I in 22 (45.8%) cases, grade II in 18 (37.5%) cases, and grade III in eight (16.7%) cases.

Comparison between controls and CSCC patients regarding MxA in tumor and stroma revealed the following: significant differences between control and CSCC groups regarding MxA positivity in both tumor and stroma were identified. All CSCC cases were positive in tumor and stroma, whereas 44. 2% of control cases showed positivity (P < 0.001 for both). A significant relationship between overexpression of MxA in CSCC epithelium and stroma was noted (P < 0.001 for both). A significant association between high H score MxA and CSCC epithelium and stroma was noted (P < 0.001 for both). All normal cases showed mild intensity, whereas 41.7% of epithelial and stromal CSCC cases show mild intensity [Table 1].
Table 1: Comparison between controls and patients with cutaneous squamous cell carcinoma regarding myxovirus resistant protein 1 in tumor and stroma

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Relation between (tumor and stroma) H scores of MxA in CSCC and clinicopathological parameters revealed the following: there was a significant association between epithelial overexpression of MxA and male sex, sun exposed area, and ulcer clinical presentation (P = 0.004, 0.02, and 0.001, respectively). Overexpression of MxA H score tumor was significantly associated with high grade and advanced stage (P = 0.02 and 0.03, respectively). Overexpression of MxA H score stroma was significantly associated with low grade and early stage (P = 0.02 and 0.02, respectively) [Table 2].
Table 2: Difference in H score regarding clinicopathological data in positive myxovirus resistant protein1 cutaneous squamous cell carcinoma cases

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Immunohistochemical expression of MxA in control and CSCC groups revealed the following: all CSCC groups showed positive MxA expression in tumor cells and surrounding stroma, whereas 44.2% of control showed positivity. All CSCC and positive control group showed cytoplasmic expression [Figure 1], [Figure 2], [Figure 3].
Figure 1: Normal skin showed mild patchy cytoplasmic expression of MxA in epidermis. Scattered view show dermal positivity (circles) (MxA IHC, ×200). IHC, immunohistochemistry; MxA, myxovirus resistant protein 1.

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Figure 2: A case of CSCC showed strong cytoplasmic MxA expression in tumor cells and the surrounding stroma (arrows) (MxA IHC, ×200). IHC, immunohistochemistry; MxA, myxovirus resistant protein 1.

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Figure 3: High-power view of previous slide demonstrating strong cytoplasmic MxA expression in tumor cells and the surrounding stroma (arrow) (MxA IHC, ×400). IHC, immunohistochemistry; MxA, myxovirus resistant protein 1.

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  Discussion Top


CSCC is a malignancy of epidermal keratinocytes that exhibits various degrees of differentiation [1]. It is the second most common cutaneous malignancy [2].

MxA, which is a major antiviral factor against a wide spectrum of RNA viruses, such as influenza virus [6], has also the potential importance in tumorigenesis and metastasis as well as in the treatment and prognosis of different cancers, such as head and neck squamous cell carcinoma, follicular lymphoma, CSCC, renal cell carcinoma, and melanoma [7]. As a key cytokine induced by types I and III IFNs, MxA is thought to be a useful biomarker for monitoring IFN activity and predicting clinical efficacy during IFN therapy in patients bearing certain types of cancer. Expression of IFN-regulated genes (including, Mx1) in squamous cell carcinoma was identified. The expression of IFN-regulated genes correlated with the extent of the lesional immune-cell infiltrate. Immunohistological examinations confirmed the expression of IFN-regulated genes in association with a CXCR3+ cytotoxic inflammatory infiltrate on the protein level. These findings support the concept that IFN-associated host responses could play an important role in tumor immunosurveillance in the skin [8]. This study aims to assess the role of MxA in CSCC. This study was done on 48 cases with CSCC and 43 age- and sex-matched apparently healthy participants.

To the best of our knowledge, there are scarce numbers of studies concerned with evaluation of the role of MxA in squamous cell carcinoma in English literation. In the current study, all CSCC cases showed positivity of MxA in the epithelium and the surrounding stroma. It could be explained by viral infections may play a role in the development of CSCC, as human papilloma virus infection has been implicated in the development of CSCC [9]. MxA is an important antiviral factor. These findings may also be owing to the expression of different mutant MxA forms. Seven point mutations affecting the structure and function of MxA could play important role in tumorigenesis and cancer development, suggesting a strong association between MxA and CSCC [5]. However, 44.2% of control cases showed positivity for MxA. It could be explained by, MxA could be involved in mitosis and transporting proteins or vesicles throughout the cell (vesicle trafficking) [10]. In the current study, overexpression of MxA in tumor and surrounding stroma was significantly noted in CSCC in comparison with control groups. It could be explained by overexpression of signal transducer and activator of transcription 1 (STAT1), which is an upstream mediator of IFN-signaling. Type I and type III IFNs bind to their cognate receptors and activate expression of MxA via the Janus-activated kinase/signal transducer and activator of transcription 1 (JAK/STAT1) pathway [4]. Upregulation of STAT1 gene expression is associated with head and neck squamous cell carcinoma (HNSCC) development. STAT1 activity functions as an oncogene mediating immune escape, cancer cell proliferation, and invasion in HNSCC [11]. In the present study, a significant overexpression of MxA in epithelial and stromal of CSCC was demonstrated. These results agreed with Wenzel et al. [12] who showed moderate to strong expression of the MxA protein in CSCC. Moreover, Song et al. [13] found that MxA is highly expressed in breast cancer.

In contrast, Calmon et al. [14] found that MxA has antitumor role in head and neck squamous cell carcinoma. This is explained by MxA promotes cell death induced by apoptotic stimuli. This provides an important link between MxA silencing and tumorigenesis. Brown et al. [15] found that low expression of MxA was noted in human prostatic cancer cell, and tissue loss of MxA expression results in increased metastasis. This could be explained by downregulation of MxA owing to a general decrease in IFN expression and signaling in prostate cancer. Davis et al. [16] found that IFN-α-induced endogenous MxA has antitumor and antimetastasis effect in hepatocellular carcinoma, as exposure of cancer cells to IFNs activates the Janus-activated kinase/signal transducer and is an activator of transcription signaling pathway.

A significant relationship between the overexpression of MxA and the male sex of the patients was also noted. Thorough search in English literatures failed to find a similar relationship. In contrary to our results, Shanora and colleagues found that androgen decreased expression of MxA, which could be explained by MxA is serum regulated and downregulated by androgen through affecting genes involved in these processes such as JAK/STAT signaling cascade [15]. Further studies are recommended to elicit the exact relationship between MxA and male gender of the patients. A significant positive correlation between MxA H score (epithelial and stromal) and sun exposed areas of CSCC cases was also noted. It could be explained by ultraviolet light induces type I IFN production in the skin through potentiation of stimulator of interferon genes (STING)-dependent activation of the immune signaling transcription factor interferon regulatory factor 3 (IRF3) in response to cytosolic DNA and cyclic dinucleotides in keratinocytes and other human cells [17].

Tumor MxA overexpression had a significant relationship with high-grade and advanced stage of CSCC. The relationship between MxA overexpression in tumor of CSCC and grade and stage was not previously done. However, MxA overexpression was correlated with tumor grade in breast carcinoma with poor prognosis patients [18]. In this study, stromal MxA overexpression had significant relationship with low grade and early stage of CSCC cases. This could be explained by MxA inhibits motility and invasiveness of cancer cells and is responsible for significant Gl cell cycle arrest, resulting in possible increase in p53 and apoptosis [19]. This raises an important question regarding the role of MxA in tumor and surrounding stroma.


  Conclusion Top


The role of MxA is complex and controversial in tumor and stroma of CSCC. Tumor MxA expression is responsible for tumor progression; however, stromal expression is responsible for regression. Further studies are recommended to identify the exact role of MxA in CSCC.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

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Heppt MV, Leiter U, Steeb T, Amaral T, Bauer A, Becker JC, et al. S3 guideline for actinic keratosis and cutaneous squamous cell carcinoma - short version, part 1: diagnosis, interventions for actinic keratoses, care structures and quality-of-care indicators. J Dtsch Dermatol Ges. 2020; 18:275-294. doi: 10.1111/ddg.14048. PMID: 32130773.  Back to cited text no. 1
    
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Kim YA, Lee HJ, Heo SH, et al. MxA expression is associated with tumor-infiltrating lymphocytes and is a prognostic factor in triple-negative breast cancer. Breast Cancer Res Treat 2016; 156:597–606.  Back to cited text no. 5
    
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Li Q, Wu H, Liao W et al. A comprehensive review of immune-mediated dermatopathology in systemic lupus erythematosus. J Autoimmun 2018; 93:1–5.  Back to cited text no. 8
    
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Chen Y, Zhang L, Graf L et al. Dynamics of dynamin-like MxA revealed by single-molecule FRET. Nat Commun 2017; 8:1–1.  Back to cited text no. 10
    
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Ryan N, Anderson K, Volpedo G, et al. STAT1 inhibits T-cell exhaustion and myeloid derived suppressor cell accumulation to promote antitumor immune responses in head and neck squamous cell carcinoma. Int J Cancer 2019; 146:1717–1729.  Back to cited text no. 11
    
12.
Wenzel J, Tomiuk S, Zahn S, et al. Transcriptional profiling identifies an interferon-associated host immune response in invasive squamous cell carcinoma of the skin. Int J Cancer 2008; 123:2605–2615.  Back to cited text no. 12
    
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Song IH, Kim YA, Lee H, et al. Retained or altered expression of major histocompatibility complex class I in patient-derived xenograft models in breast cancer. Immunol Res 2019; 67:469–477.  Back to cited text no. 13
    
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Calmon MF, Rodrigues RV, Kaneto CM, et al. Epigenetic silencing of CRABP2 and M×1 in head and neck tumors. Neoplasia 2009; 11:1329.  Back to cited text no. 14
    
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Brown SG, Knowell AE, Hunt A, Patel D, Bhosle S, Chaudhary J. Interferon inducible antiviral MxA is inversely associated with prostate cancer and regulates cell cycle, invasion and Docetaxel induced apoptosis. Prostate 2015; 75:266–279.  Back to cited text no. 15
    
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Davis D, Yuan H, Yang YM, Liang FX, Sehgal PB. Interferon-α-induced cytoplasmic MxA structures in hepatoma Huh7 and primary endothelial cells. Contemp Oncol 2018; 22:86.  Back to cited text no. 16
    
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Skopelja-Gardner S, An J, Sun X, et al. UV light induces acute type I interferon production in the skin and blood which is cGAS dependent. Arthritis Rheumatol 2019; 71:111.  Back to cited text no. 17
    
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Sistigu A, Yamazaki T, Vacchelli E, et al. Cancer cell–autonomous contribution of type I interferon signaling to the efficacy of chemotherapy. Nat Med 2014; 20:1301.  Back to cited text no. 18
    
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    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

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