Petcharin Srivatanakul

Genetic and environmental determinants of risk for cholangiocarcinoma via Opisthorchis viverrini in a densely infested area in Nakhon Phanom, northeast Thailand

Infection with Opisthorchis viverrini (OV) is associated with cholangiocarcinoma. OV is common in northeast Thailand, but less than 10% of the inhabitants develop cholangiocarcinoma. Animal experiments suggest that OV infection alone does not cause cholangiocarcinoma, and thus other environmental and genetic factors may play a role in causation. We conducted a population‐based case‐control study in which sex, age and place of residence were matched individually. Polymorphisms of GSTM1 and GSTT1 alone were not associated with risk for cholangiocarcinoma, while an elevated level of antibodies against OV (ELISA) ≥0.200 was the strongest risk indicator (odds ratio as compared to that <0.200 = 27.09 [95% confidence interval (CI): 6.30–116.57]. Compared to subjects who had a normal antibody range and the wild‐type GSTM1 gene, those who had elevated antibodies had higher odds ratios of 12.32 (95% CI: 1.60–94.85) for wild‐type GSTM1 and 23.53 (95% CI: 4.25–130.31) for the null variant thereof, respectively. Past and current regular drinkers of alcohol had higher risk [odds ratio = 5.39 (95% CI: 1.11–26.06) and 4.82 (95% CI: 1.29–18.06), respectively]. Eating fermented products was an independent risk factor. Smokers or consumers of fermented fish had substantially increased risk if they were past or current drinkers. Infection with OV correlates strongly with cholangiocarcinoma, susceptibility to which may be possibly associated with GSTM1 polymorphism. Alcohol may affect metabolic pathways of endogenous and exogenous nitrosamines.

The role of infection by opisthorchis viverrini, hepatitis B virus, and aflatoxin exposure in the etiology of liver cancer in thailand. A correlation study

The incidence of the two principle types of liver cancer (hepatocellular carcinoma and cholangiocarcinoma) in five different areas of Thailand was compared with the prevalence of exposure to the main risk factors in samples of the population. Cholangiocarcinoma showed striking variations in incidence, which correlated closely with markers of exposure to the liver fluke, Opisthorchis viverrini. However, there was little geographic variation in incidence of hepatocellular carcinoma or in prevalence of the major risk factors (chronic carriage of hepatitis B virus and exposure to aflatoxin), and apparently there was little relationship between them. Cancer 68:2411–2417, 1991.

Colon and Rectum Cancer in Thailand: An Overview

Cancers of the colon and rectum are rare in developing countries, in contrast to the high incidence rates in countries of Europe, North America, Australia and Japan. Significant differences also exist within continents. Colorectal cancer mortality and incidence rates have decreased in the USA. However, the incidence in Japan and Thailand is rising, probably due to the acquisition of Western lifestyle. Incidence also increases with age: carcinomas are rare before the age of 40 years except in individuals with genetic predisposition or predisposing conditions. The incidence rate of colorectal cancer in Thailand is low when compared with other countries. It is the third in frequency in males after liver and bile duct and lung cancers, and the fifth after cancers of the cervix, breast, liver and bile duct and lung for females. The highest incidence for both sexes is seen in Bangkok. The number of cases of colorectal cancer in both sexes is increasing and will probably exceed that of lung cancer in the next decade. Thus, we are planning to have colorectal cancer screening programme. We should pay more attention on primary and secondary prevention to control colorectal cancer in Asian countries.

CYP1A1, GSTM1, and GSTT1 Polymorphisms, Smoking, and Lung Cancer Risk in a Pooled Analysis among Asian Populations

To evaluate the roles of CYP1A1 polymorphisms [Ile462Val and T6235C (MspI)] and deletion of GSTM1 and GSTT1 in lung cancer development in Asian populations, a pooled analysis was conducted on 13 existing studies included in Genetic Susceptibility to Environmental Carcinogenesis database. This pooled analysis included 1,971 cases and 2,130 controls. Lung cancer risk was estimated as odds ratios (OR) and 95% confidence intervals (95% CI) using unconditional logistic regression model adjusting for age, sex, and pack-year. The CYP1A1 6235C variant was associated with squamous cell lung cancer (TC versus TT: OR, 1.42; 95% CI, 0.96-2.09; CC versus TT: OR, 1.97; 95% CI, 1.26-3.07; Ptrend = 0.003). In haplotype analysis, 462Val-6235T and Ile-C haplotypes were associated with lung cancer risk with reference to the Ile-T haplotype (OR, 3.41; 95% CI, 1.78-6.53 and OR, 1.39; 95% CI, 1.12-1.71, respectively). The GSTM1-null genotype increased squamous cell lung cancer risk (OR, 1.36; 95% CI, 1.05-1.77). When the interaction was evaluated with smoking, increasing trend of lung cancer risk as pack-year increased was stronger among those with the CYP1A1 6235 TC/CC genotype compared with those with TT genotype (Pinteraction = 0.001) and with the GSTM1-null genotype compared with the present type (Pinteraction = 0.08, when no genotype effect with no exposure was assumed). These results suggest that genetic polymorphisms in CYP1A1 and GSTM1 are associated with lung cancer risk in Asian populations. However, further investigation is warranted considering the relatively small sample size when subgroup analyses were done and the lack of environmental exposure data other than smoking. (Cancer Epidemiol Biomarkers Prev 2008;17(5):1120–6)

Natural variations of copper and sulfur stable isotopes in blood of hepatocellular carcinoma patients

Significance In cancer, the metabolism of copper and sulfur are dysregulated, leading to deleterious side effects. These issues are commonly addressed by studying the variations of concentrations of the elements, but here we have used, for the first time to our knowledge, copper and sulfur stable isotope compositions variations, using methods widespread in Earth sciences. We show that in hepatocellular carcinomas patients, blood copper and sulfur are enriched in light isotopes compared with control subjects. These isotopic signatures are not compatible with a dietary origin, but rather reflect the massive reallocation in the body of copper immobilized within cysteine-rich proteins such as metallothioneins. We also propose that sulfur isotope compositions could serve to track sulfur originating from tumor-derived sulfides. The widespread hypoxic conditions of the tumor microenvironment can impair the metabolism of bioessential elements such as copper and sulfur, notably by changing their redox state and, as a consequence, their ability to bind specific molecules. Because competing redox state is known to drive isotopic fractionation, we have used here the stable isotope compositions of copper (65Cu/63Cu) and sulfur (34S/32S) in the blood of patients with hepatocellular carcinoma (HCC) as a tool to explore the cancer-driven copper and sulfur imbalances. We report that copper is 63Cu-enriched by ∼0.4‰ and sulfur is 32S-enriched by ∼1.5‰ in the blood of patients compared with that of control subjects. As expected, HCC patients have more copper in red blood cells and serum compared with control subjects. However, the isotopic signature of this blood extra copper burden is not in favor of a dietary origin but rather suggests a reallocation in the body of copper bound to cysteine-rich proteins such as metallothioneins. The magnitude of the sulfur isotope effect is similar in red blood cells and serum of HCC patients, implying that sulfur fractionation is systemic. The 32S-enrichment of sulfur in the blood of HCC patients is compatible with the notion that sulfur partly originates from tumor-derived sulfides. The measurement of natural variations of stable isotope compositions, using techniques developed in the field of Earth sciences, can provide new means to detect and quantify cancer metabolic changes and provide insights into underlying mechanisms.

GSTM1 and CYP1A1 polymorphisms, tobacco, air pollution, and lung cancer: a study in rural Thailand.

Objectives: The Lampang Province is situated in the northern region of Thailand. Incidence rates of lung cancer are high for Asian standards, particularly in women. This study was conducted to quantify the risk of lung cancer associated with exposures prevalent in the area and to investigate possible interactions with genetic susceptibility. The presence of several large open-cast coal mines from 1955 close to electricity-generating plants was a particular focus of concern. Methods: Two-hundred and eleven cases of primary lung cancers diagnosed in 1993 to 1995 and residents in the province were recruited at the Lampang Provincial Hospital (main referral center for treatment of the disease). Two sets of controls, frequency-matched to the cases by sex and age, were recruited (a) from the resident population (202 interviewed) and (b) from patients admitted to the hospital for diseases predominantly unrelated to tobacco smoking (211 interviewed). Sociodemographic information, complete residential history, and characteristics of the household (place of cooking, cooking fuel, and heating fuels), occupational history, and history of tobacco smoking were obtained by interview. Cases and controls (∼50% of the population-based series) provided a blood sample. A point source air pollution exposure index was calculated for each village/township reported in residential histories based on the linear distance from the Mae Moh Center (the area of the electricity-generating plants), the year-specific gaseous (SO2 and NO2) or total suspended particulate emissions from the Mae Moh Power Plant, and the percentage of wind from the center. Odds ratios (OR) for the disease associated with categorical variables were estimated within unconditional logistic regression. Extraction of genomic DNA and genotyping of variants in CYP1A1 and GSTM1 were conducted to assess the extent of modification of risk by these genes that are involved in the metabolism of polycyclic aromatic hydrocarbons, a common component of the exposures. Results: Overall, there was no evidence of relevant differences in the socioeconomic level of the three groups. The two control sets were similar with respect to lifelong tobacco habit and were subsequently pooled in analyses. Never-smokers were 7% of men and 33% of women. Smoking of local traditional products unfiltered and high in tar content is a common habit in the rural female population. ORs associated with smoking increased with duration of the habit and average daily amount, being 4.9 [95% confidence interval (95% CI), 2.5-9.7] for smokers of ≥7 cigarettes/d and 3.3 (95% CI, 1.7-6.2) for duration of 41 years or longer compared with nonsmokers. Smoking of local products was associated with an independent OR of 3.1 (95% CI, 1.7-5.6) adjusted for lifelong cumulative amount of tobacco smoked. Although most smokers had the habit for at least 16 years, the daily consumption was low compared with Western standards. Other potential sources of exposure to lung carcinogens (emission from the power-generating plants and domestic burning of coal and wood for cooking and heating) were not associated with increased risk of lung cancer. None of the three polymorphisms examined increased the risk of lung cancer or modified the risk associated with tobacco smoking. Conclusion: In this rural population, 96% of male and 64% of female lung cancer incidence were explained by tobacco smoking. None of the potential sources of air pollution deriving from the combustion of coal and wood, or polymorphisms in the CYP1A1 gene or deletion of the GSTM1 had an effect on the risk of lung cancer, either together or separately. (Cancer Epidemiol Biomarkers Prev 2006;15(4):667–74)

The p53 codon 72 polymorphism in Thai nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) is a serious cancer in Thailand. To address on the genetic risk factor for NPC, we investigated association between the p53 codon 72 polymorphism (Pro/Arg) and NPC susceptibility in the Thai. The genotype frequency of this polymorphism in 102 NPC patients and 148 age-matched healthy controls was determined by using a PCR-RFLP assay. No statistically significant difference in the overall genotype frequencies or allele frequencies between cases and controls was observed. Among NPC patients, no statistical significant difference in p53 genotype frequencies between sex, histological types and clinical stages was observed. When cases and controls were categorized into 3 groups of age, >40, >45 and >50 years, the frequencies of p53 genotype in the cases were significantly different from those of the controls (P<0.05). The p53 Pro homozygotes with age of >40 years had twofold to threefold higher risk to develop NPC and the risk was increased with age (Odds Ratio (OR) increased from 2.01 to 2.63 by age >40 to >50 years). Individuals of age >40, >45 and >50 years with an Arg/Arg genotype and of age >45 and >50 years with combined Arg/Arg or Arg/Pro genotype had lower risk to develop NPC than those with Pro/Pro genotype (P<0.05). Collectively, we suggest that the p53 gene polymorphism may associate with NPC susceptibility in Thai population, particularly the Pro/Pro genotype carriers with age of >40 years. The detection of this p53 polymorphism may be a useful tool for screening of early stage and diagnosis of NPC.

DNA methylation differences in exposed workers and nearby residents of the Ma Ta Phut industrial estate, Rayong, Thailand

BACKGROUND Adverse biological effects from airborne pollutants are a primary environmental concern in highly industrialized areas. Recent studies linked air pollution exposures with altered blood Deoxyribo-nucleic acid (DNA) methylation, but effects from industrial sources and underlying biological mechanisms are still largely unexplored. METHODS The Ma Ta Phut industrial estate (MIE) in Rayong, Thailand hosts one of the largest steel, oil refinery and petrochemical complexes in south-eastern Asia. We measured a panel of blood DNA methylation markers previously associated with air pollution exposures, including repeated elements [long interspersed nuclear element-1 (LINE-1) and Alu] and genes [p53, hypermethylated-in-cancer-1 (HIC1), p16 and interleukin-6 (IL-6)], in 67 MIE workers, 65 Ma Ta Phut residents and 45 rural controls. To evaluate the role of DNA damage and oxidation, we correlated DNA methylation measures with bulky DNA and 3-(2-deoxy-β-D-erythro-pentafuranosyl)pyrimido[1,2-α]purin-10(3H)-one deoxyguanosine (M(1)dG) adducts. RESULTS In covariate-adjusted models, MIE workers, compared with rural residents, showed lower LINE-1 (74.8% vs 78.0%; P < 0.001), p53 (8.0% vs 15.7%; P < 0.001) and IL-6 methylation (39.2% vs 45.0%; P = 0.027) and higher HIC1 methylation (22.2% vs 15.3%, P < 0.001). For all four markers, Ma Ta Phut residents exhibited methylation levels intermediate between MIE workers and rural controls (LINE-1, 75.7%, P < 0.001; p53, 9.0%, P < 0.001; IL-6, 39.8%, P = 0.041; HIC1, 17.8%, P = 0.05; all P-values vs rural controls). Bulky DNA adducts showed negative correlation with p53 methylation (P = 0.01). M(1)dG showed negative correlations with LINE-1 (P = 0.003) and IL-6 methylation (P = 0.05). CONCLUSIONS Our findings indicate that industrial exposures may induce alterations of DNA methylation patterns detectable in blood leucocyte DNA. Correlation of DNA adducts with DNA hypomethylation suggests potential mediation by DNA damage.

Malondialdehyde-deoxyguanosine adducts among workers of a thai industrial estate and nearby residents

Background Humans living near industrial point emissions can experience high levels of exposures to air pollutants. Map Ta Phut Industrial Estate in Thailand is the location of the largest steel, oil refinery, and petrochemical factory complexes in Southeast Asia. Air pollution is an important source of oxidative stress and reactive oxygen species, which interact with DNA and lipids, leading to oxidative damage and lipid peroxidation, respectively. Objective We measured the levels of malondialdehyde–deoxyguanosine (dG) adducts, a biomarker of oxidative stress and lipid peroxidation, in petrochemical workers, nearby residents, and subjects living in a control district without proximity to industrial sources. Design We conducted a cross-sectional study to compare the prevalence of malondialdehyde-dG adducts in groups of subjects experiencing various degrees of air pollution. Results The multivariate regression analysis shows that the adduct levels were associated with occupational and environmental exposures to air pollution. The highest adduct level was observed in the steel factory workers. In addition, the formation of DNA damage tended to be associated with tobacco smoking, but without reaching statistical significance. A nonsignificant increase in DNA adducts was observed after 4–6 years of employment among the petrochemical complexes. Conclusions Air pollution emitted from the Map Ta Phut Industrial Estate complexes was associated with increased adduct levels in petrochemical workers and nearby residents. Considering the mutagenic potential of DNA lesions in the carcinogenic process, we recommend measures aimed at reducing the levels of air pollution.

Association between EBNA2 and LMP1 subtypes of Epstein-Barr virus and nasopharyngeal carcinoma in Thais

BACKGROUND Nasopharyngeal carcinoma (NPC) is an Epstein-Barr virus (EBV)-associated malignancy that is common in Thailand. The association of the EBV nuclear antigen 2 gene (EBNA2) and the latent membrane protein 1 gene (LMP1) with NPC is unclear. OBJECTIVE To determine the association between EBNA2 and LMP1 subtypes of EBV and NPC in Thais. STUDY DESIGN EBV DNA from blood samples of 75 Thai patients with NPC and 44 age-matched controls was examined by PCR for EBNA2 subtypes (EBV type 1 and type 2) and LMP1 subtypes (wild-type LMP1 (wt-LMP1) and LMP1 containing a 30-bp deletion (del-LMP1)). RESULTS The frequencies of EBV type 1 in cases (96.0%) and controls (97.7%) were similar. The frequency of del-LMP1 in cases (58.7%) was higher than in controls (36.4%) (p=0.023). Among NPC patients, the frequency of del-LMP1 in stages III and IV (65.6%) was higher than in stages I and II (18.2%) (p=0.006). By analyzing nucleotide sequence of the LMP1 region, a new subtype, Thai1, was identified. CONCLUSION LMP1 subtypes of EBV may be involved in NPC susceptibility. The del-LMP1 variant may be a useful molecular marker for early detection, diagnosis and prognosis of Thai patients with NPC.