子宮頸病變患者周邊血液淋巴球之人類乳突瘤病毒16/18 DNA存在與其組織p16INK4a蛋白表現之相關性研究

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Title:	子宮頸病變患者周邊血液淋巴球之人類乳突瘤病毒16/18 DNA存在與其組織p16INK4a蛋白表現之相關性研究 Relationships between the presence of HPV 16/18 DNA in peripheral blood lymphocytes and p16INK4a protein expression in human papillomavirus-associated neoplastic cervical lesions
Authors:	彭雅汶 Ya-Wen Peng
Contributors:	中山醫學大學：毒理學研究所;李輝;邱慧玲
Keywords:	子宮頸癌;人類乳突瘤病毒;p16 INK4a 蛋白 cervical cancer;human papillomavirus;p16 INK4a protein
Date:	2003
Issue Date:	2010-06-04T09:18:14Z (UTC)
Abstract:	子宮頸癌是目前全球女性常見的一種癌症，發生率之高儼然成為危害全球女性同胞健康的頭號殺手。雖然子宮頸癌是種可以先預防、初期治癒率也相當高的癌症，但是在台灣由於子宮頸抹片之篩檢率相當低，因此其死亡率仍是居高不下，顯示子宮頸癌之防治與早期診斷仍為尚待加強之重要課題。許多相關研究都明確證實人類乳突瘤病毒是子宮頸癌發生的罪魁禍首，也有許多研究找出了高危險型HPV E6及E7蛋白的致癌分子機轉。其中HPV E7 所導致p16INK4a 蛋白在子宮頸組織的高度表現情形，早已被證實可以作為子宮頸癌的診斷指標。本實驗室之前在肺癌以及子宮頸癌病患的血球玻片上測得HPV DNA的表現，而且在肺癌病患血球細胞中偵測到HPV DNA可以作為肺癌病患的風險評估。本研究擬分別探討高度鱗狀上皮內病灶(high-grade squamous intraepithelial lesion; HSIL)與早期子宮頸癌(early invasive cervical cancer; EICC)病患血液中HPV DNA 的表現，與子宮頸組織發生病變的相關性。因此選用36位HSIL與32位EICC患者，分別以nested-PCR分析血液中與病變組織HPV的感染率，並且以PCR-RFLP的實驗方法作再次確認，結果發現EICC病患的PBL(peripheral blood lymphocyte)中HPV DNA表現率為53.1% ，HSIL患者也有25% 偵測出HPV DNA。在PBL可偵測到HPV DNA的26位病人，在子宮頸組織全都可以測到HPV DNA (P < 0.001)，其一致性高達100%。另外，感染HPV的病患在PBL中也可測到HPV E7 mRNA的表現(在HSIL為33.3%, P = 0.012 ; 在EICC為47.1%, P = 0.002)，顯示在周邊血液HPV的感染的確可以反映出子宮頸癌組織的感染情形。並且在研究中也證實，當子宮頸組織受到HPV感染後，HPV E7致癌蛋白會促使p16INK4a蛋白的表現量增加，同時也觀察到在子宮頸病變組織HPV DNA以及E7 mRNA的表現都和p16INK4a蛋白表現量成正相關性，尤其以HSIL患者為明顯。而在HSIL患者血液中有HPV 16或是HPV 18 存在的9位病人中，p16INK4a蛋白全都是高度表現 (P = 0.009)，在EICC病患感染HPV且同時表現p16INK4a蛋白則有94.1% (P = 0.228)。顯示在PBL 偵測出HPV DNA與p16INK4a蛋白的表現有顯著相關性，而且在病變早期關係更為顯著。同樣地，HPV E7 mRNA與p16INK4a蛋白的表現也有相似的情形 (在HSIL 為100%, P = 0.009 ; 在EICC為92.3%, P = 0.458) 。由以上結果得知，偵測PBL中HPV 16 或18 DNA的存在，不僅可以反映組織中HPV 的感染情形，亦跟組織中p16INK4a蛋白的高度表現有正相關性。 Cervical cancer has been the major cause for cancer-related morbidity and mortality for females worldwide. The significant contribution of human papillomavirus to cervical carcinogenesis has also been intensively investigated, and many reports have shown that p16INK4a may serve as an early biomarker of cervical cancer since p16INK4a was up-regulated by E2F through Rb inactivation by HPV 16 E7. Our preliminary data showed that HPV DNA signals were detected in peripheral blood lymphocytes (PBL) of lung cancer and cervical cancer patients by in situ hybridization. In this study, we attempted to investigate the correlation between p16INK4a protein expression and human papillomavirus infection in PBL and tumor tissues from patients with high-grade squamous intraepithelial lesion (HSIL) or early invasive cervical cancer (EICC) to verify the presence of HPV DNA in PBL plays a crucial role in cervical carcinogenesis. Paired tissues and peripheral blood samples from 68 patients, including 36 HSIL patients and 32 EICC patients, were collected for studies. The presence of HPV type 16/18 DNA and E7 mRNA in tissue and PBL was examined by nested-PCR and PCR-RFLP, respectively. Results showed that the presence of HPV 16 and/or 18 DNA in tissues was significantly correlated with those in PBL from all cases (P = 0.005 for HPV 16, P = 0.022 for HPV 18 and P < 0.001 for HPV 16 or 18), especially those 26 patients with HPV DNA in both PBL and tissues. This result revealed that HPV infection in cervical tissues can be indirectly indicated by a direct detection of HPV DNA in blood circulation. Moreover, our data supported that HPV E7 mRNA expression was respectively correlated with Rb and p16 immunostainings in cervical tissues from patients with HSIL and EICC (P < 0.001). This result seemed to confirm the previous reports showing that p16 up-regulation may be mediated through Rb inactivation by HPV 16/18 E7. In addition, the positive p16 immunostaining was positively correlated with the presence of HPV 16 or 18 DNA in cervical tissues of HSIL (P < 0.001), but not in EICC. Interestingly, a positive correlation was also observed in PBL of HSIL (P = 0.044 for HPV 16, P = 0.009 for HPV 16 or 18), but still not in EICC. In conclusion, the present study clearly revealed the positive association between the presence of HPV 16 or 18 DNA in PBL and p16 overexpression in tissues of patients with cervical lesions.
URI:	http://140.128.138.153:8080/handle/310902500/1558
Appears in Collections:	[醫學分子毒理學研究所] 博碩士論文

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