Investigation Of HLA Allele Susceptibility And Traditional Chinese Medicine Constitution In Guangdong Han Patients With Nasopharyngeal Carcinoma

Nasopharyngeal carcinoma is a common malignant tumor in southern China, especially in Guangdong Province.According to statistics, the incidence of nasopharyngeal carcinoma (NPC) in the Guangdong region is30-50/10people.Surgery has difficulty in excising malignant tumor completely because of its deep location and being surrounded by a lot of important organizations and tissues.In our country, more than90%of nasopharyngeal carcinoma (NPC) is non-keratinising undifferentiated carcinoma, which is extremely sensitive to radiation therapy.Therefore, radiotherapy (RT) is the primary treatment for nasopharyngeal carcinoma (NPC) due to its anatomical and biological characteristic.While the therapeutic result of patients with early nasopharyngeal carcinoma treated with radiotherapy alone is desirable, with the5-year survival rate more than90%, most patients with NPC are diagnosed only when the tumor has reached an advanced stage (stages III and IV) due to the non-specific nature of the nasal and aural early symptoms, the difficulty of making a clinical examination of the nasopharynx and the deep location of NPC.The treatment effect of patients with locally advanced nasopharyngeal treated "with radiotherapy alone is poor.Radiotherapy is local treatment which cannot effectively kill distant metastasis cancer cells, whereas chemotherapy is systemic treatment which has disadvantage in killing distant metastasis cancer cells.Radiotherapy with concomitant chemotherapy has increased the overall survival of patients with local advanced nasopharyngeal carcinoma, however, the side effects of chemoradiation are serious, causing poor tolerability and lower completion rate of chemotherapy.The exact etiology of nasopharyngeal carcinoma is unclear, which is considered to be related to the physical and chemical environment factors, viral factors, genetic factors and so on.Early linkage analysis on Chinese sib Pair studies of NPC suggested the association of susceptibility HLA haplotypes with NPC development.HLA genes are highly polymorphic and encode for human leukocyte antigen (HLA) molecules that are essential for the presentation of foreign antigens to the immune system.In addition, HLA gene can be genetic mark to study hereditary disease.Constitution is formed on the basis of the growth and development process. This special status often determines the specificity of their physiological responses. The polymorphism of TCM constitution are similar to the HLA gene polymorphism.Purpose:To find out the susceptible HLA genes and TCM constitution of the Guangdong Han patients with nasopharyngeal carcinoma.Method:Patients with nasopharyngeal carcinoma(114cases) were randomly selected in Guangdong Han population from the period2012/1to2012/10, and121case of healthy volunteers served as the control group.SBT gene sequencing technologies were applied to analyse the HLA-A/B/DRB1gene polymorphism in both groups, in order to evaluate the association between the susceptible HLA genes.In addition,100cases, aged from18to65, were randomly selected respectively from each group, who agreed to take the TCM constitution questionnaire.The questionnaire was TCM constitution scale, which originated from the TCM Constitution Classification and Evaluation Criteria.The NPC group was told to complete the questionnaire according to their actual1-year situation one month before the diagnosis of NPC; while the control group according to their actual situation within the previous year.The investigation of HLA polymorphism susceptible in Guangdong Han people with nasopharyngeal carcinoma use the following statistical methods:Hardy-Weinberg Equilbrium test was performed using Alequin version3.11.Frequency distribution of genetypes and alleles between the two groups was compared using Pearson X2tests, Yates test or Fisher’s exact test was used when necessary.Odds Ratios (ORs) and95%Confidence Intervals were calculated using Woolf method.Data were presented as means±standard deviation (SD) or as percentages for discrete variables.The independent sample T Test was used to compare the differences in quantitative traits between the two groups.Chi-Square test was used for the comparisions of the proportions between the two groups.All tests were two sided and the statistical significance level a was0.05.Statistical analysis was performed using SPSS version13.0(SPSS, Inc.) and Excel.The investigation of Chinese Medicine Constitution in Guangdong Han people with nasopharyngeal carcinoma use the following statistical methods:Chi-Square test was used for the comparisions of the proportions between two groups.All tests were two sided and the significance level a was0.05.Boferronni method was used to adjust the statistical significance level for the comparitions of Qi deficiency constitution and Qi depression constitution (α=0.025).Result:HLA-A allele frequency between the two groups are compared, the HLA-A*11:01gene frequencies in the NPC group were significantly statistical different to the normal control group (A*11:01,0.257vs0.357, OR=0.570,95%CI=0.340-0.956, x2=4-57, P=0.033).The remaining HLA-A allele frequency differences has no statistically significant between them.HLA-B allele frequency between the two groups are compared, the B*58:01gene frequency in the NPC group were significantly statistical different to the normal control group (0.121vs0.064, OR=2.0879,95%CI=1.041-4.186, X2=4.17, P=0.036).The remaining HLA-B allele frequency differences has no statistically significant between them.HLA-DRB1allele frequency between the two groups are compared, the DRB1*03:01> DRB1*04:03gene frequency in the NPC group were significantly statistical different to the normal control group (DRB1*03:01:0.116vs0.047, OR=2.087,95%CI=1.311-6.020, X2=7459, P=0.006; DRB1*04:03:0.022vs0.0064, OR=0.324,95%CI=0.114-0.923, x2=4.57, P=0.028).The remaining HLA-DRB1allele frequency differences has no statistically significant between them. Compare HLA-A、B、DRB1allele frequencies between EBV-EA-IgG negative and positive NPC patients, frequencies of HLA-A*33:03, B*38:01and DRB1*01:01has statistically significant differences between two groups.(A*33:03: OR=1.282,95%CI:1.149-1.429, x2=6.130, P=0.013; B*38:01, OR=6.222,95%CI:4.075-9.501, P=0.029;DR1*01:01, OR=6.222,95%CI:4.075-9.501, P=0.029), and the remaining HLA allele frequency differences has no statistically significant between them.Compare HLA-A、B、DRB1allele frequencies between EBV DNA negative and positive NPC patients, frequencies of HLA-B*15:19and B*46:01has statistically significant differences between two groups.(B*15:19, OR=5.600,95%CI:3.764-8.331, P=0.036; B*46:01:OR=6.222,95%CI:1.240-12.575, x2=5.976, P=0.015), and the remaining HLA allele frequency differences has no statistically significant between them.The TCM constitution of the patients with nasopharyngeal carcinoma in Guangdong Han people were mostly Qi deficiency typeand Qi depression type before they became sick.Conclusion:HLA-B*58:01and DRB1*O3:O1NPC may be the susceptible genes to NPC in Guangdong Han people. HLA-A*11:01and DRB1*04:03may be the protective genes to NPC in Guangdong Han people.Furthermore, Qi deficiency and Qi depression constitution may be the susceptible TCM constitution to NPC in Guangdong Han people...