Human Papillomaviruses Testing and DNA Microarray Analyses in the Management of Patients with Cervical Neoplasm

• Main Authors: ANGEL CHAO, 趙安琪
• Other Authors: Chyong-Huey Lai
• Format: Others
• Language: en_US
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/40921829942360999873

博士 === 長庚大學 === 臨床醫學研究所 === 94 === Extensive epidemiologic and molecular research has strongly linked anogenital infection of certain human papillomaviruses (HPVs) to the development of cervical neoplasia and cervical cancer. Currently, HPV DNA detection has played a triage role in the management of atypical squamous cells of undetermined significance (ASCUS), primary screening with cytology for the detection of cervical cancer and cervical intraepithelial neoplasia (CIN), and follow-up in a variety of clinical settings. In this study, I analyzed consecutively a series of 2,154 patients undergoing conization of the cervix. Patients who were diagnosed with cervical cancer or CIN 1, had a hysterectomy within 12 weeks after conization, or had no follow-up data were excluded. The remaining 765 patients were analyzed. Of the 765 patients, 279 had dysplasia at the margins of the conization or endocervix (group A) while 486 were free of neoplasia at both the margins and endocervix (group B). The 3-year cumulative rate of residual/recurrent high-grade CIN was 10.3%. Our results indicated that HPV follow-up status, margin status and follow-up cervical cytology were significant predictors for residual/recurrent high-grade CIN. This study suggested that HPV DNA testing is useful in the follow-up of these patients and in the understanding of the natural history after conization for high-grade CIN. In a parallel attempt to understand molecular mechanisms for the different clinical features between adenocarcinoma/adenosquamous carcinoma (AC/ASC) and squamous carcinoma (SC) of the uterine cervix, I analyzed gene expression profiles of different histological subtypes of cervical cancer. Cancer specimens and surrounding normal tissue counterparts were separately collected from cervical cancer patients undergoing type III radical hysterectomies. DNA microarray analyses indicated that genes including carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5), tumor-associated calcium signal transducer 1 (TACSTD1), S100 calcium binding protein P (S100P), and mesothelin (MSLN), were upregulated in AC. Contrarily, genes involved in the epidermal differentiation complex, such as S100 calcium binding protein A9 (S100A9) and annexin A8 (ANXA8), were upregulated in SC. Cross-validation of the results using an independent comparable group of patients with known long-term outcomes showed that the correlation between the six selected differentially-expressed genes and histology was highly significant. Furthermore, using a multivariate Cox proportional hazards regression analysis, I found CEACAM5 and TACSTD1 to be highly significant prognostic factors. The combination of cDNA microarray, RTQ-PCR, and immunohistochemical results of this study has clearly defined different gene profiles for AC and SC. Moreover, TACSTD1 expression may be a novel prognostic factor for poor outcome. To investigate patient responses to cancer treatment at the transcriptional level, I analyzed serial changes of gene expression profiles in peripheral blood of cervical cancer patients (bulky stage IB –IVA squamous cell carcinomas) undergoing concurrent chemoradiation (CCRT). Total RNAs at four time points (before CCRT treatment, one week after the first course of chemotherapy, one week and one month after the completion of CCRT) were analyzed using DNA microarray. Fifty-three genes that were significantly overexpressed during CCRT were categorized into two groups by hierarchical clustering. Group 1, consisting of nine genes including S100A9, was upregulated during CCRT. Group 2 (44 genes) was upregulated after treatment during which patients had been severely myelosuppressed. In Group 2, fetal and embryonic hemoglobin genes (HBG2 and HBZ) along with adult hemoglobin genes (HBA2 and HBB), red blood cells isozyme [carbonic anhydrase I (CA1)], and enzymes involved in heme biosynthesis [aminolevulinate, delta-, synthase 2 (ALAS2)] were overexpressed. Cancer-related genes, cathepsin D (CTSD) and activin A receptor, type I (ACVR1) were significantly upregulated in anemic patients in CCRT. Immunoglobulin kappa variable 1D-13 (IGKV1D-13) was significantly downregulated in patients with grade 3/4 hematologic toxicity. Therefore, declining hemoglobin may reactivate fetal and embryonic hemoglobin genes as well as the enzymatic activities of erythrocytes and cancer-related genes in cervical cancer patients during CCRT. Our results suggested that the downregulation of IGKV1D-13 may predict hematologic toxicity in cervical cancer patients during CCRT.