人類卵巢原發性鱗狀細胞癌（SCC）是相當罕見的。大多數案例被認為是由於卵巢畸胎瘤、卵巢纖維上皮瘤或子宮內膜異位症所導致的惡性病變，但此癌症的確實病因仍然未明。人類乳突病毒（HPV）感染是誘導腫瘤形成的關鍵因素，尤其是子宮頸癌。根據上述發現，因此，本研究的目的是探討成熟畸胎瘤(MCT)受人類乳突病毒感染，與卵巢惡性轉化鱗狀細胞癌形成的關聯性。研究中使用石蠟包埋之檢體，包括高雄榮民總醫院 25 年來所收集得之四個 SCC-MCT 組織與子宮頸至卵巢間之相鄰組織、12 個良性畸胎瘤卵巢組織與 11 個良性卵巢組織；並包括兩例子宮頸鱗狀癌組織作為控制組。以 HPV 16/18 外殼蛋白或 E6 蛋白之抗體進行免疫組織化學染色(IHC)；或利用 HPV6/11，16/18 和 31/33 等三組基因型探針進行原位雜交(ISH）檢測 HPV 病毒織感染。免疫組織化學染色結果顯示四例 SCC-MCT 組織樣本與兩例子宮頸鱗狀癌控制組皆有受到 HPV 感染。重要的是，也可在 SCC-MCT 樣本之相鄰組織(子宮頸至卵巢間組織)中檢測出 HPV 病毒，顯示病毒顆粒可能藉由上行性路徑，通過子宮頸/子宮頸內膜/子宮內膜輸卵管至卵巢。原位雜交(ISH)結果則顯示所有 SCC-MCT組織樣本皆為 HPV 16/18 型陽性、並有兩例 SCC-MCT 組織樣本呈現 HPV31/33 陽性，而並未在 SCC-MCT 組織樣本中檢測出 HPV 6/11 型病毒。不過，與 SCC-MCT 組織樣本相比之下，皮樣組織和非皮樣組織之 HPV 病毒檢出率低，強烈暗示 HPV 病毒之感染可能與正常卵巢組織或良性卵巢畸胎瘤無關聯性。

Primary squamous cell carcinoma (SCC) of the ovary in humans is rare. Most cases represent a malignant transformation of ovarian teratoma, Brenner tumor, or endometriosis. The etiology of this cancer remains largely unknown. Human papillomavirus (HPV) infection is a critical factor that induces tumor formation of cervical cancer. Based on the above findings, this study aimed to evaluate the association of HPV with malignant transformation of mature cystic teratoma (MCT) into the SCC of ovary. The specimens used in this study included formalin-fixed paraffin-embedded SCC-MCT tissues and their adjacent tissues from the cervix to ovaries of four patients. A total 12 cases of benign teratoma ovarian tissues (dermoid tissues) and 11 cases of benign non-teratoma ovarian tissues (non-dermoid tissues) were collected and examined. Two squamous carcinoma tissues of the cervix were used as control samples. HPV was detected by immunohistochemistry (IHC) with antibodies against either HPV capsid or E6 of HPV type 16/18, and by in situ hybridization (ISH) with three sets of genotyping probes of HPV types 6/11, 16/18 and 31/33. The results of IHC analysis revealed the HPV infection in the four cases of SCC-MCT and the two control cervical cancer samples. Importantly, HPV
was also detected in adjacent reproductive tissues of the SCC-MCT cases, suggesting that viral particles might spread through an ascending tissue route from the fallopian tubes, endometrium, endocervix and cervix to the ovaries. ISH results revealed the presence of HPV type 16/18 in all SCC-MCT cases and HPV type 31/33 in two, with no HPV type 6/11 in any SCC-MCT cases. However, when compared with the SCC-MCT cases, the
lower detection rates of HPV in dermoid cysts and nondermoid tissues suggested that HPV viral infection might not be associated with normal ovarian tissues or benign ovarian teratomas.

目 錄
論文審定書﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍
誌謝﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ i
中文摘要﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ ii
英文摘要﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ iii
目錄﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ iv
圖次目錄﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ vi
表次目錄﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ vii
補充資料﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ viii
1. Introduction﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 1
1.1 HPVs historical background﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 2
1.2 Classification of human papillomavirus and Mechanisms of HPVs carcinogenesis﹍﹍﹍﹍﹍﹍ 2
1.3 Human ovarian cancer﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 5
2. Materials and Methods﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 8
2.1 Human tissue samples﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍8
2.2 Cell type for experiment﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 8
2.3 Immunohistochemistry (IHC) ﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍9
2.4 In situ hybridization (ISH) ﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 9
2.5 Statistical analysis﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 10
3. Results﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍10
3.1 Four SCC-MCT cases﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 10
3.2 IHC of HPV﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍12
3.3 ISH of HPV﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍15
4. Discussion﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 15
5. Conclusions and Limitation﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 18
6. Future Research Directions﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 18
7. Figures and Figure Legends﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 19
References﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍24
Supplemental materials﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 30
圖 次 目 錄
Figure 1.The location in the squamous epithelium of the main stages of the papillomavirus life cycle﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 19
Figure 2. Human papillomavirus (HPV) diagnosis by immunohistochemical (IHC) analysis﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 20
Figure 3. Positive IHC data of Case 1 SCC-MCT and adjacent tissues﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 21
Figure 4.Human papillomavirus (HPV) detection in dermoid and nondermoid ovarian tissues﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 22
Figure 5. Human papillomavirus (HPV) detection by in situ hybridization (ISH) ﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍23
表 次 目 錄
Table 1. Clinical characteristics and outcome of the four cases of squamous cell carcinoma with mature cystic teratoma (SCC-MCT) ﹍ 12
Table 2. Human papillomavirus detection by immunohistochemistry in tissue biopsies of cancer patients﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 14
Table 3. HPV detection by immunohistochemical analysis in dermoid and nondermoid cyst tissues﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 14
補 充 資 料
Supplementary Figure 1. Schematic presentation of the HPV genome﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 30
Supplementary Table 1. The World Health Organization (WHO) classification of ovarian tumors﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 31
Supplementary Table 2. Classification of sex cord–stromal tumors﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 32
Supplementary Table 3. WHO classification of germ cell tumors of the ovary﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 33
Clinical data with image and pathological figures in one case in our study﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍ 34

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