Nm23-H1, P53及MMP-2作為早期子宮頸癌生物因子的臨床應用探討

The Clinical Significance of Nm23-H1, P53 and MMP-2 as Biomakers at Early Stage Cervical Cancer Patients

鄭義銅(Yi-Torng Tee)

指導教授 ： 林隆堯

中山醫學大學/醫學院/醫學研究所/博士(2008年)

https://doi.org/10.6834/CSMU.2008.00072

摘要

子宮頸癌是台灣婦女發生率第一位的癌症, 每年共有6300人罹患子宮頸癌. 我們的研究主要在探討p53, mmp-2及nm23-H1這三種代表性的基因在早期子宮頸癌生物因子的臨床意義. 本研究分別進行了nm23-H1, p53 及MMP-2三個基因表現蛋白的免疫組織化學染色. nm23-H1及p53基因表現蛋白部分是由福馬林固定的石蠟包埋蠟磈,製成約5微米( micrometer; mm)檢體切片共49x2=96 片; 而另一部分MMP-2的免疫組織化學染色則是從Zymed laboratories 製作成的組織晶片Tissue Array上使用定量軟體Pros Plus 4.5.1 image analysis program,及進行RT-PCR 實驗.
我們的結果顯示nm23-H1 及MMP-2基因蛋白是細胞質染色, 而p53基因蛋白是細胞核染色. 早期子宮頸癌病患的nm23-H1與p53的基因表現是不一致的, 即p53的表現並不會促使nm23-H1也併表現, 反之亦然. 所有臨床病理因子, 包括了年齡( 大於等於50歲vs 小於50歲), 組織分化程度(低及中等分化vs高度分化), 基質侵犯深度( 超過等於一半vs 少於一半), FIGO分期( 第一vs 第二期)和組織細胞型態( 鱗狀上皮細胞vs腺癌)中只有組織分化程度及基質侵犯深度有統計上顯著差異( P<0.05), 即組織分化較差及基質侵犯深度超過等於一半者比組織分化較好及基質侵犯深度沒有超過等於一半者的病人較有可能會有骨盆腔淋巴結轉移的機會. 我們發現nm23-H1及p53蛋白的sensitivity, specificity, positive predictive value, negative predictive value分別為nm23-H1 的91.7%, 13.5%, 25.6%, 83.3%, 32.7% 及p53的 66.7%, 51.4%, 30.8%, 82.6%,意即nm23-H1及p53蛋白準確性並不高.MMP-2基因的RNA表現性在各組SCC,高度鱗狀上皮內病變及正常組織的表現情形如下: SCC組中90% 表現及高度鱗狀上皮內病變組中有70%表現. MMP-2的基因表現強度IOD在鱗狀上皮侵犯癌(SCC)組的表現顯著高於Normal, Low-grade CIN 及High-grade CIN組( P<0.001) 而High-grade CIN組其mmp-2基因蛋白表現也顯著高於Normal, Low-grade CIN, 但normal組及Low-grade CIN組這兩組的mmp-2基因蛋白表現在統計上沒有顯著差異. 這結果顯示mmp-2基因調控與子宮頸癌的癌化有關, 而且這可能與MMP-2蛋白分解酵素能分解上皮下基底膜及能侵入基質有關.

關鍵字

Nm23-H1 ； P53 ； MMP-2 ； 子宮頸癌

並列摘要

Cervical cancer is the most common type of cancer among women in our country Taiwan, accounting for 6300 new case diagnosed annually. We have chosen representative tumor gene p53, MMP-2 and nm23-H1 to study their expression in cervical cancer at the early stage. There were two parts in our study, immunoreactivities of nm23-H1 and p53 proteins had been evaluated in 49 patients of early stage cervical cancer in the first part while the correlation of matrix metalloproteinase-2 (MMP-2) protein with cervical carcinogenesis had been studies in the second part of our study via tissue array using image analysis program Pro Plus 4.5.1. and reverse-transcription polymerase chain reaction (RT-PCR).
We found nm23-H1 and MMP-2 immunoreactivity was cytoplasmic , however, p53 was nuclear. Immunoreactivities of nm23-H1 and p53 products are different or disassociated. Subgroups analysis revealed that the independent variables predictive of lymph nodes metastases include grade of tumor cell differentiation (P<0.05) and stromal invasion (P<0.05). Age, FIGO stage, and histopathology were not predictive of lymph node metastasis , and neither nm23-H1 nor p53 expression was associated with lymph node metastasis in our 49 cervical cancer patients. The sensitivity, specificity, PPV, NPV, and accuracy of nm23-H1 and p53 for the prediction of lymph nodes metastases in cervical cancer were 91.7%, 13.5%, 25.6%, 83.3%, 32.7% and 66.7%, 51.4%, 30.8%, 82.6%, and 55.1%, respectively . This results show that Nm23-H1 and p53 are dissociated and not good predictors of lymph node metastases in early-stage cervical cancer patients, but stromal invasion and cell differentiation can predict lymph node metastasis. We further found the mRNA expression of MMP-2 to be higher in most SCCs (9/10 samples) and high-grade CINs ( 7/10 samples) but lower in normal tissues. The IOD of MMP-2 was significantly higher in high-grade CIN than in normal and low-grade CIN tissue cores (P<0.001 for both) and significantly higher in SCC than in high-grade CIN tissue cores (P< 0.001). This results show that MMP-2 upregulation confers on tumor cells the ability to degrade the subepithelial basement membrane and subsequently invade the cervix

並列關鍵字

Nm23-H1 ； P53 ； MMP-2 ； cervical cancer

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