Human leukocyte antigen and esophageal ... - Wiley Online Library

Diseases of the Esophagus (2006) 19, 238–240

Original article

Blackwell Publishing Asia

Human leukocyte antigen and esophageal cancer in the northwestern region of Iran J. Eivazi-Ziaei,1 S. Dastgiri,2 J. Majidi,3 J. Vaez,1 I. Asvadi,1 A. Mahmoudpour4 1

Tabriz University Hematology Oncology Research Centre, 2Epidemiology Department, 3Immunology Department, Center for Applied Drug Research, Tabriz University of Medical Sciences, Tabriz, 51665-139, Iran

4

SUMMARY. Esophageal squamous cell carcinoma is the 6th most commonly occurring cancer worldwide. A relationship between HLA A1 and B40 and esophageal cancer was described in patients examined in China. The aim of this study was to investigate the relation of HLA class 1 and esophageal carcinoma in the northwestern region of Iran. Using specific monoclonal antibodies, different human leukocyte antigens (HLA) were quantified in 100 patients suffering esophageal carcinoma in Tabriz, a major city located in the Northwestern region of Iran. These data were compared to those of 100 healthy matched individuals as a control group from the same region. HLA B14 and A24 were increased and showed statistically significant correlation in squamous cell carcinoma. These findings may also indicate the association between genetic factors and esophageal carcinoma. Further studies are suggested for detecting correlation of HLA and esophageal carcinoma in other regions. KEY WORDS: carcinoma, esophagus, HLA.

INTRODUCTION Esophageal squamous cell carcinoma (SCC) is the 6th most commonly occurring cancer worldwide.1,2 The areas with higher incidence include Northern provinces of Iran and certain regions of China, South America and Africa.2–5 According to the Iranian Institute of Cancer, 9% of all cancers and 27% of gastrointestinal cancers have been shown to be esophageal carcinoma.2 The high frequency of esophageal cancer has been continuously reported from the northern region of Iran since 1980.4 Both genetic and environmental factors should be considered when a disease etiology is investigated.6 Several reports from different regions of the world with high-risk populations, including the northern provinces of Iran, indicate a positive family history among patients with esophageal cancer.7,8 While reports from China which has extraordinarily higher rates of SCC have suggested a Mendelian mode of transmission, the higher significance of overall allelic loss in patients with a positive family history Address correspondence to: Jamal Eivazi-Ziaei, MD., Associate Professor of Internal Medicine, hematologist oncologist, Tabriz University Hematology Oncology Research Centre, Tabriz University of Medical Sciences, Tabriz, 51665-139, Iran. Email: [email protected] 238

suggest the presence of an inherited tumor suppressor gene on 13q.9 The level of HLA-DRB1*0901 AF was increased in patients with esophageal cancer compared to healthy individuals.7 A relationship between HLA A1 and B40 and esophageal cancer was described in patients examined in the Linxian area of China;10 this may suggest the possibility of a genetically determined risk. The aim of this study was to investigate the relation of HLA class 1 in the northwestern region of Iran, an area with prevalent occurrence of esophageal carcinoma. The primary objective of this investigation was to determine the HLA specificities in esophageal carcinoma.

METHODS AND PATIENTS One hundred adult patients with esophageal carcinoma and 100 healthy individuals selected randomly as a control group matched with cases by ethnicity and residence status from the same region, were studied in Shahid Ghazi hematology and oncology hospital and outpatients clinic. Inclusion criteria were those patients pathologically proven to suffer from esophageal carcinoma. In cases lacking appropriate control individuals, certain patients were excluded

© 2006 The Authors Journal compilation © 2006 The International Society for Diseases of the Esophagus

Esophageal carcinoma in Iran 239

from the study. An informed consent was obtained from all patients and controls for blood sampling. The university ethical committee approved this study. The healthy individuals were all donor candidates for patients with renal failure in desperate need of kidney transplantation. The patients were introduced to the transplantation immunology laboratory for typing of HLA. A volume of 5.0 mL defibrinated blood diluted 1 : 1 with Hanks balanced salt solution buffer (HBSS), pH 7.2 was added to 3.0 mL of separating medium (ficol-hypaqe, D = 1.077). After centrifugation at 1000 g for 20 min, lymphocyte enriched Buffy coat layers were washed with HBSS. One µL of product, equivalent to 2000–5000 lymphocytes was added to micro plate wells coated with different anti-HLA monoclonal antibodies. After incubation for 20– 30 min at 37°C with gentle shaking, 5.0 µL of rabbit complement was added at each well and incubated for 1–1.5 h under the same conditions. Finally, eosin and formalin were added to the wells. After incubating for 24 h at 0°C, live and dead lymphocytes were differentiated using an inverted microscope in order to compare positive (ALG) and negative (pooled AB) controls. The results of patients’ HLA were compared to those of 100 controls. The data were analyzed using descriptive statistics and two proportions statistical test (Z-test) to compare HLA proportions between cases and controls.

Table 1 Basic characteristics of cases Age groups 30 –39 40 – 49 50 –59 60 – 69 70 –79 80+ Gender Male Female Residence status (by province) East Azerbaijan West Azerbaijan Ardebil Kurdistan

Frequency 1 7 23 35 22 12

Percent 1 7 23 35 22 12

52 48

52 48

81 15 3 1

81 15 3 1

of all types of HLA. In contrast, analysis of SCC data showed significant differences only in A24 and B14 compared to those of controls.

DISCUSSION The surveys in Iran and China suggest that the natural history of esophageal cancer starts with esophagitis which, in a few individuals, progresses to atrophy and dysplasia of the epithelium and finally to cancer.11,12 Consumption of wheat flour, exposure to residues from opium pipes, drinking hot tea, chewing nass (a mixture of tobacco, lime, ash, and some other ingredients), and bread are potentially the causal agents of esophageal cancer in Iran.2,5,13–15 Several studies have demonstrated the involvement of some nutrients, vitamins, vegetables, fruits and socioeconomic status in the etiology of SCC.5,11,14,16 –19 However, the disease may still be seen in the same family at a higher rate than expected by chance alone.15,16 Due to the obstacles in early diagnosis and poor efficacy of treatment, the 5-year survival rate of SCC is usually less than 10%. According to Yan Nie et al., human SCC is developed through a multistep process. Delineation of the molecular mechanisms involved in this process will not only provide biomarkers for early detection, but also enable us

RESULTS Table 1 describes the demographic specificities of patients and controls. The highest occurrence of disease was observed among patients 60–69 years of age. SCC included 93% of all esophageal cancer in the region compared to 7% of adenocarcinoma cases. Occurrence of SCC was significantly higher in the lower third of the esophagus (49.5%, P < 0.001). Table 2 presents the statistical comparison of HLA distribution between adenocarcinoma and SCC groups to those of control individuals. Apart from B14, adenocarcinoma cases were shown to be statistically different from control groups in terms Table 2 Human leukocyte antigens (HLA) in esophageal carcinoma SCC

Adenocarcinoma

Control

P-value

HLA

Frequency

%

Frequency

%

Frequency

%

SCC versus control

Adenocarcinoma versus Control

A10 A24 B5 B14 B35 BW4 BW6 CW4

14 23 57 2 5 23 6 26

3.2 5.3 13.2 0.5 1.2 5.3 1.4 6

0 2 3 2 1 2 0 2

0 6.5 9.7 6.5 3.2 6.5 0 6.5

10 10 58 5 35 77 59 70

1.3 0.3 7.8 0.7 4.7 10.4 7.9 9.4

NS 0.02 NS 0.001 NS NS NS NS

0.02 0.0006 0.002 NS 0.02 0.002 0.00002 0.02


240 Diseases of the Esophagus

to improve our treatment procedures.1 Major histocompatibility complex (MHC) is a genetic term describing alleles encoding antigens which principally determine the fate of graft transplantation. In many species, the MHC is also described by an additional term such as HLA in humans.7 According to our data, SCC included 93% of esophageal carcinomas in the northwestern region of Iran with the highest prevalence in the lower third of the esophagus. HLA B14 and A24 provided significant relation to squamous cell carcinoma in this study. Chinese investigators described correlation of HLA B40 and HLA A1 with SCC.10 This might have been correlated to variations of HLA in different regions, or to different natural histories of disease in China. Although HLA B5 was more prevalent in SCC and adenocarcinoma, it was shown to be significant in adenocarcinoma only. This might, of course, be because of the small number of adenocarcinoma cases in this study. HLA A24 was reported to be significantly higher in both adenocarcinoma and SCC. Further studies should be undertaken in other regions with appropriate numbers of patients to confirm these findings. There may be an internal environment susceptible to malignancy and a genetic component in the patients’ families, which supports the concept that heredity may play an important role in the pathogenesis. It is still controversial whether or not HLA antigen expression in carcinomas correlates with development of carcinoma and its prognosis.7 June Lin and coworkers demonstrated a relatively higher ratio of chromosomal abnormality in cancerous relatives as compared to the general population. The inheritance patterns however, clearly do not fit in a simple Mendelian pattern.7 Our study is another that may show a correlation of genetic factors with SCC; however, further studies are required to delineate the relationship of other inherited factors and esophageal cancer.

CONCLUSION We conclude that HLA A24 is significantly higher in SCC patients in the north-western region of Iran. Further investigations with more advanced methodologies, and larger sample size in other regions are suggested to clarify the possible correlation of SCC to HLA and other inherited factors.

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