HOTAIR與NR5A2基因多型性與南台灣口腔潛在惡性病症癌化、口腔癌發生和預後之間的相關性分析

Associations between Malignant Transformation of Oral Potentially Malignant Disorders, Oral Cancer Development and Prognosis, and Polymorphisms in HOTAIR and NR5A2 genes in Southern Taiwan Populations.

盧韋智(Wei-Chih Lu)

指導教授 ： 林英助

高雄醫學大學/口腔醫學院/口腔衛生學系碩士班/碩士(2017年)

摘要

背景:細胞異常增生與DNA甲基化是癌症發生和預後變差的重要危險因子，HOTAIR (Hox transcript antisense intergenic RNA) rs920778基因多型性的表現異常上調與使DNA甲基化和人類癌症的不良預後相關，導致癌症加重；NR5A2 (The orphan nuclear receptor) rs3790844表現升高與癌細胞生長，增殖和分化關係密切。近幾年來口腔癌的發生率與死亡率仍然在持續增加，但HOTAIR rs920778和NR5A2 rs3790844基因在口腔癌在潛在惡性病症癌化、口腔癌發生和預後扮演的角色仍然不清楚。
研究目的:我們將收集健康對照組、口腔潛在惡性病症和口腔癌患者來分析HOTAIR rs920778和NR5A2 rs3790844對口腔癌發生和預後變差與口腔潛在惡性病症癌化潛力增加的基因易感性。
研究方法:口腔癌和口腔潛在惡性病症 (oral potentially malignant disorder; OPMD) 等研究對象從高雄醫學大學附設中和紀念醫院科口腔顎面外科收集。疾病診斷皆經過口腔組織病理切片和化學染色確認；健康對照組為40歲 (含) 以上。經簽署人體試驗同意書之後，才開始進行資料的收集，研究對象基本資料和潛在干擾因子則以標準化結構式問卷加以收集；抽取5-8 ml的周邊血液作為萃取DNA所需；HOTAIR和NR5A2基因多型性採用即時聚合酶鏈式反應結合螢光探針，結果經獨立判讀後判別，將所有資料經過建檔與連結後，以統計軟體SPSS版本19.0進行卡方檢定、邏輯式迴歸、生存曲線分析以及COX比例風險模式來分析其關聯性。
結果:研究對象有口腔癌患者267人，口腔潛在惡性病症患者160人，以及健康對照組276人納入分析。HOTAIR rs920778與NR5A2 rs3790844基因多型性之頻率分佈在口腔癌和健康對照組之間和口腔潛在惡性病症患者與健康對照組之間皆未達統計上顯著差異，但NR5A2基因多型性頻率分佈在口腔潛在惡性病症患者與口腔癌患者 (X2 = 5.37，p = 0.002)之間有顯著差異。邏輯式回歸分析發現NR5A2 rs3790844基因多型性是增加口腔潛在惡性病症轉變成口腔癌癌化潛力的危險因子(AA vs. GG+AA，odds ratio (OR) = 2.31，95%CI = 1.02-5.24)，但是沒有明顯與口腔癌(AA vs. GG+AA，OR = 2.28，95%CI = 0.88-5.90)或口腔惡性病症(AA vs. GG+AA， OR = 0.54，95%CI = 0.12-2.30)的發生有相關性。在口腔癌患者之生存曲線分析顯示口腔癌患者的整體存活率（overall survival）在不同HOTAIR基因多型性分組間沒有明顯差異 (log-rank = 0.084，p = 0.772)，但在不同NR5A2基因多型性分組間有明顯差異 (log-rank = 3.887，p = 0.049)；口腔癌患者的無病生存率在不同HOTAIR或不同NR5A2基因多型性間皆沒有明顯差異
(HOTAIR: log-rank=0.309，p = 0.578；NR5A2: log-rank = 0.472，p=0.492)。最後經過多變項COX比例風險模式分析，結果顯示HOTAIR基因多型性與口腔癌患者的整體存活率沒有明顯相關 (HOTAIR: adjusted hazard ratio (aHR) = 1.20，95%CI = 0.37-3.89)，但是NR5A2基因多型性經backward stepwise多變項COX比例風險模式分析，結果顯示其(AA vs. GG+GA基因型：aHR = 2.10，95%CI = 1.00-4.37) 與口腔癌患者較低的整體存活率有關。此外HOTAIR與NR5A2基因多型性皆與口腔癌患者的無病累積存活率的改變無明顯相關 (HOTAIR:aHR = 0.66，95%CI = 0.16-2.73；NR5A2:aHR = 0.78，95%CI = 0.28-2.18)。
結論: 研究結果發現NR5A2 rs3790844基因多型性與口腔潛在惡性病症癌化潛力增加有顯著相關性，也與口腔癌患者較低的整體存活率有關，但與口腔潛在惡性病症發生或口腔癌的發生或口腔癌患者的無病存活率變差無明顯相關。而HOTAIR基因多型性皆與以上所觀察到的結果無明顯相關性

關鍵字

HOTAIR與NR5A2基因多型性 ； 口腔癌 ； 口腔潛在惡性病症癌化

並列摘要

Background: The out of control in cell proliferation was a hall marker of cancer. The activation of Hox transcript antisense intergenic RNA (HOTAIR) and Nuclear Receptor Subfamily 5 Group A Member 2 (NR5A2) genes played important roles of cancer cell proliferation, invasion, progression, and metastasis as well as poor survival in a variety of human cancers. In Taiwan, the incidence rate and mortality rate was still increased in past decade. However, the roles of HOTAIR and NR5A2 A>G polymorphisms in the development of oral cancer and oral potentially malignant disorder (OPMD), and prognosis of oral cancer patients remains unclear.
Objective: Our aims were to investigate the association between polymorphisms in HOTAIR rs920778T>C, NR5A2 rs3790844A>G genes in OPMD development, OSCC development, malignant potential from OPMD to oral cancer, survival in oral cancer patients.
Material and Methods: Patients with OPMD and OSCC were collected from Kaohsiung Medical University Hospital. Healthy controls were collected from people who were invited from Community Health examinations. After the inform consent was obtained. Potential confounding factors and demographic variables were collected using a standardized structure questionnaire and 8 ml of peripheral blood was collected for genomic DNA extraction. Genetic polymorphisms were determined using real-time PCR TaqMan probe method. SPSS statistical software version 19.0 was used to analyze the association.
Result: More than 80% of the study subjects were already recruited in our previous studies. After the IRB was obtained, total of 267 oral cancer patients, 160 OPMD patients and 276 healthy controls were recruited into this study. The frequency distributions of NR5A2 rs3790844 polymorphisms was significant different between oral cancer group and OPMD group (χ2 = 5.37， p = 0.002) but no such association was found for HOTAIR rs920778 polymorphisms. Polymorphisms in HOTAIR genes were not associated with the development of oral cancer but NR5A2 polymorphisms were associated with OPMD turn into oral cancer. The logistic regression analysis only showed that NR5A2 polymorphisms were significantly associated with the OPMD turn into oral cancer (GG+GA vs.AA，odds ratio (OR) = 2.31，95%CI = 1.02-5.24) but no such association was found for HOTAIR rs920778 polymorphisms.. Survival curve analysis showed that HOTAIR rs920778 were not associated with the overall survival of oral cancer patients but NR5A2rs3790844 polymorphisms were (HOTAIR: log rank = 0.084，p = 0.772; NR5A2: log-rank = 3.887，p = 0.049). Disease free survival curve analysis showed that both HOTAIR rs920778 and NR5A2rs3790844 polymorphisms were not associated with the disease free survival of oral cancer patients (HOTAIR: log-rank = 0.309，p = 0.578; NR5A2: log-rank = 0.472，p = 0.492). However, multivariate Cox proportional hazard model analysis showed that HOTAIR rs920778 and NR5A2 rs3790844 polymorphisms were not associated with the low overall survival in oral cancer patients (HOTAIR: adjust hazard ratio (aHR) = 1.20，95%CI = 0.37-3.89；NR5A2:aHR = 2.05，95%CI = 0.98-4.29). According to backward stepwise analysis, HOTAIR rs920778 polymorphisms were not associated with the low overall survival in oral cancer patients but NR5A2 rs3790844 polymorphisms were (HOTAIR: aHR = 1.20，95%CI = 0.37-3.89；NR5A2: aHR = 2.05，95%CI = 0.98-4.29).Both HOTAIR rs920778 and NR5A2rs3790844 polymorphisms were not associated with low disease free survival in oral cancer patients (HOTAIR: aHR = 0.66，95%CI = 0.16-2.73；NR5A2: aHR = 0.78，95%CI = 0.28-2.18).
Conclusion: These results suggested that NR5A2 rs3790844 polymorphisms can confer the genetic susceptibility to the high potential of malignant transformation from OPMD to oral cancer, but no such relationship was found for HOTAIR polymorphisms.

並列關鍵字

Polymorphisms in HOTAIR and NR5A2 genes ； Oral Potentially Malignant Disorders ； Oral Cancer

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