Pituitary Adenylate Cyclase-activating Polypeptide Inhibits Pacemaker Activity of Colonic Interstitial Cells of Cajal

This study aimed to investigate the effect of pituitary adenylate cyclase-activating peptide (PACAP) on the pacemaker activity of interstitial cells of Cajal (ICC) in mouse colon and to identify the underlying mechanisms of PACAP action. Spontaneous pacemaker activity of colonic ICC and the effects of PACAP were studied using electrophysiological recordings. Exogenously applied PACAP induced hyperpolarization of the cell membrane and inhibited pacemaker frequency in a dose-dependent manner (from 0.1 nM to 100 nM). To investigate cyclic AMP (cAMP) involvement in the effects of PACAP on ICC, SQ-22536 (an inhibitor of adenylate cyclase) and cell-permeable 8-bromo-cAMP were used. SQ-22536 decreased the frequency of pacemaker potentials, and cell-permeable 8-bromo-cAMP increased the frequency of pacemaker potentials. The effects of SQ-22536 on pacemaker potential frequency and membrane hyperpolarization were rescued by co-treatment with glibenclamide (an ATP-sensitive K+ channel blocker). However, neither N(G)-nitro-L-arginine methyl ester (L-NAME, a competitive inhibitor of NO synthase) nor 1H-[1,2,4]oxadiazolo[4,3-alpha]quinoxalin-1-one (ODQ, an inhibitor of guanylate cyclase) had any effect on PACAP-induced activity. In conclusion, this study describes the effects of PACAP on ICC in the mouse colon. PACAP inhibited the pacemaker activity of ICC by acting through ATP-sensitive K+ channels. These results provide evidence of a physiological role for PACAP in regulating gastrointestinal (GI) motility through the modulation of ICC activity.

Ewing's Sarcoma of the Stomach; Rare Case of Ewing's Sarcoma and Suggestion of New Treatment Strategy

Ewing's sarcoma is a neoplasm of the undifferenciated small round cells, which generally affects the bone and deep soft tissues of children and adolescents. We present a case of gastric Ewing's sarcoma; a 35-year-old female who had no symptoms. While she was at a routine medical checkup, a protruding mass in her gastric antrum was incidentally found on esophagogastroduodenoscopy. Endoscopic ultrasonogram showed a submucosal mass on the same lesion and a laparosopic wedge resection was done. Pathologic gross findings showed a granular grape appearance tissue and histoloigc examination revealed a small round cell tumor with CD 99 immunoexpression positive. In general, a combined modality therapy for Ewing's sarcoma such as surgical resection with chemotherapy, is accepted as an effective method. However, this patient had no adjuvant chemotherapy after surgery and she has no recurrence for eleven months.

Ewing's Sarcoma of the Stomach; Rare Case of Ewing's Sarcoma and Suggestion of New Treatment Strategy

Ewing's sarcoma is a neoplasm of the undifferenciated small round cells, which generally affects the bone and deep soft tissues of children and adolescents. We present a case of gastric Ewing's sarcoma; a 35-year-old female who had no symptoms. While she was at a routine medical checkup, a protruding mass in her gastric antrum was incidentally found on esophagogastroduodenoscopy. Endoscopic ultrasonogram showed a submucosal mass on the same lesion and a laparosopic wedge resection was done. Pathologic gross findings showed a granular grape appearance tissue and histoloigc examination revealed a small round cell tumor with CD 99 immunoexpression positive. In general, a combined modality therapy for Ewing's sarcoma such as surgical resection with chemotherapy, is accepted as an effective method. However, this patient had no adjuvant chemotherapy after surgery and she has no recurrence for eleven months.

Prognostic Implications of the Expression of CXCL16 in Breast Carcinoma

BACKGROUND: Of the many prognostic factors for breast cancer, the relationship between an infiltration of inflammatory cells and the prognosis is debatable. Of the chemokines affecting cancer's inflammatory reactions, chemokine (C-X-C motif) ligand 16 (CXCL16) has attracted attention for its prognostic value in many cancers, including colorectal cancer and renal cell carcinoma. But the situation for breast carcinoma is unknown. The aim of this study was to examine the relationship between the prognostic factors and the CXCL16 expression in patients with breast carcinoma. METHODS: The patients (n=106) diagnosed with invasive ductal cancer of the breast were enrolled. We reviewed the clinicopathological factors of these patients, hematoxylin and eosin stains were prepared and estrogen receptor (ER), human epidermal growth factor receptor 2 (HER2/neu) and CXCL16 immunostaining was performed. RESULTS: The ER expression was significantly correlated with age and inflammation. A CXCL16 expression was noted in 81.1% of the cases. No association was evident between a CXCL16 expression and any other parameter, including the survival rate. Multivariate analysis did not implicate ER, HER2/neu or CXCL16 as an independent prognostic factor, but the tumor size was independent predictive factor for the patient outcome. CONCLUSIONS: An inflammatory reaction mediated by CXCL16 is not associated with the prognosis of breast cancer or any clinicopathological factors.

Significance of the Expression of Cathepsins B, H, & L in Colonic Epithelial Neoplasms

BACKGROUND: Cathepsin is associated with tumorigenesis, tumor invasion and metastasis through its ability to induce degradation of extracellular matrix components. METHODS: To investigate the correlation between cathepsin expression and tumor progression, invasion depth or nodal metastasis, immunohistochemical staining for cathepsins B, H and L were done on 20 hyperplastic polyps, 48 adenomas, and 67 adenocarcinomas of the colon. Evaluation of the expression of cathepsins B, H and L was based on the percentage of neoplastic cells that stained positive for any given cathepsin. RESULTS: Cathepsin B expression was significantly higher in adenocarcinomas than adenomas (29.33 vs 5.48%), but was not associated with the degree of differentiation, depth of invasion and nodal status of the tumors. Expression of cathepsins H and L was absent or low in both adenomas and adenocarcinomas. CONCLUSIONS: We suggest that cathepsin B is involved in progression of a subset of colonic adenomas, while cathepsins H and L are not.

Squamous Cell Carcinoma of an Ileal Neobladder: A Case Report

Bladder reconstruction using bowel segments, especially the ileum, has become a realistic option for urinary diversion. There is only one prior case of squamous cell carcinoma of the ileal neobladder that has been reported in the clinical literature. Here we report a patient with a spectrum of squamous cell lesions, including squamous cell carcinoma, sarcomatoid carcinoma, squamous papilloma and squamous metaplasia that developed in the ileal neobladder. A 46-year-old woman underwent a hysterectomy, cystectomy and ileocystoplasty for tuberculosis 25 years previously complained of urinary frequency and gross hematuria for one week. A pelvic CT revealed a 6.3 cm mass in the neobladder. The histopathological examination showed an 11x8 cm polypoid fragile mass with a microscopically well-differentiated squamous cell carcinoma, squamous papilloma and non-tumor squamous metaplasia.

The Differential Expressions of the Epithelial-Mesenchymal Transition Regulator, Slug and the Cell Adhesion Molecule, E-cadherin in Colorectal Adenocarcinoma

BACKGROUND: Slug is a member of the Snail family of transcription factors, and it plays a crucial role in the regulation of the epithelial-mesenchymal transition by suppression of several epithelial proteins and adhesion molecules, including E-cadherin. METHODS: The aim of the present study was to examine the significance between the expression of Slug in colorectal adenocarcinoma (CRA) specimens and the clinicopathological parameters of CRA, as determined by immunohistochemical analysis, and to determine the correlation between the Slug and E-cadherin expressions in non-neoplastic colorectal mucosa (n=45), primary CRA (n= 109) and metastatic CRA (n=17). A semiquantitative scoring system was applied based on the intensity and extent of the positive immunohistochemical staining. RESULTS: The expressions of Slug and E-cadherin were associated with the depth of tumor invasion (pT) (p=0.019, p=0.001, respectively), and these expressions showed a significant inverse correlation (p<0.001) each other. CONCLUSIONS: Our results demonstrated a positive role for Slug in the development of CRA, and Slug is a mediator of tumor invasion in CRA. In addition, an up-regulated Slug expression is significantly correlated with the loss of an E-cadherin expression, which suggests that Slug may play some role in the epithelial-mesenchymal transition (EMT) by down-regulating the E-cadherin expression.

Papillary carcinoma arising in a struma ovarii of the complex teratoma / 대한산부인과학회지

Benign or malignant tumors that arise in a benign cystic teratoma are rare. Malignant transformation in mature cystic teratoma of ovary is reported to occur in 1~3%. Several types of malignancy can occur from any of three germ-cell layers, but the most common malignancy is a squamous cell carcinoma derived from ectoderm. We experienced a papillary carcinoma arising in a struma ovarii of the complex teratoma. 74-year-old female presented with an abdominal discomfort. A pelvic ultrasonograph and computed tomography detected a 7 x 6 cm-sized and well-circumscribed cystic mass at the anterior to the uterus. The inner part of the mass was filled with cystic fluid showing low-density, however, solid portion including fat and soft tissue accompanied by calcification was identified in more than half of the entire tumor volume. The patient underwent a bilateral salpingo-oophorectomy. On histopathologic examination, the lesion was consistent with papillary carcinoma with squamoid features arising in a struma ovarii of the mature cystic teratoma. We herein report a case of the very rare papillary carcinoma arising in a struma ovarii of the complex teratoma and review the available literature.

Expression of Osteopontin, ZO-1 and E-cadherin in Adenoma and Adenocarcinoma of the Colon

Background : The expressions of osteopontin (OPN), zonula occludens-1 (ZO-1) and E-cadherin, known as cell adhesion-associated substances, were examined in adenoma and adenocarcinoma of the colon. The relationship of their expressions with clinicopathologic factors was examined to investigate the roles of these proteins in the development, invasion or metas- tasis of colon adenocarcinoma. Methods : The expressions of OPN, ZO-1, and E-cadherin were examined in 54 cases of adenoma and 67 cases of adenocarcinoma of the colon by immunohistochemical staining. Results : The expression of OPN in colon adenocarcinoma correlated with staging (p=0.012) and distant metastasis (p=0.021). The expression of ZO-1 was closely related with tumor cell differentiation (p<0.001), and the reduced expression of E-cadherin was associated with tumor cell differentiation (p=0.05) and lymph node metastasis (p<0.001). Co-expression of ZO-1 and E-cadherin was significantly associated with tumor cell differentiation, and the expressions of ZO-1 and E-cadherin were reduced or lost in all cases (5 cases) of poorly differentiated adenocarcinoma. Conclusions : Our data suggest that OPN is involved in the process of invasion and metastasis of colon adenocarcinoma, and ZO-1- and E-cadherin-mediated cell adhesion may play an important role in the differentiation of colon adenocarcinoma.

Degenerative Change of Cerebellar Purkinje Cells by Harmaline Treatment / 체질인류학회지

The indole alkaloid harmaline has been to cause tremor and ataxia, and produce cerebellar neurotoxicity in rat. Degeneration of Purkinje cell alligned in narrow parasagittal bands result from excitation of inferior olivary nucleus in harmaline-treated rats. The objective of this study was to investigate the hypothesis that excitation of climbing fiberinduced by harmaline mediates Purkinje cell injury or degeneration. For this purpose, the inferior olive of rats was chemically ablated by using 3-acetyl pyridine, a neurotoxic chemical, and cerebellar damage followed by administration of harmaline was analyzed using immunohistochemical markers for neurons, glial cells. The results demonstrated that a subset of Purkinje cell in the vermis and paravermis degenerated after harmaline treatment, but harmaline produced little or no Purkinje cell degeneration after inferior olivary ablation. These results suggested that harmalineinduced activation of inferior olivary neurons may lead to release of glutamate from climbing fiber synaptic terminal distributed over the Purkinje cells, and may lead to cytotoxic degeneration of Purkinje cells.

Radiation-Induced Apoptosis of Lymphocytes in Peripheral Blood / 대한방사선종양학회지

PURPOSE: This study quantitatively evaluated the apoptosis in human peripheral blood lymphocytes using flow cytometry, and investigated the possibility of using this method, with a small amount of blood, and the time and dose dependence of radiation-induced apoptosis. MATERIALS AND METHODS: Peripheral blood lymphocytes were isolated from the heparinized venous blood of 11 healthy volunteers, 8 men and 3 women, with each 10 ml of blood being divided into 15 samples. The blood lymphocytes were irradiated using a linear accelerator at a dose rate of 2.4 Gy/min, to deliver doses of 0.5, 1, 2 and 5 Gy. The control samples, and irradiated cells, were maintained in culture medium for 24, 48 and 72 hours following the irradiation. The number of apoptotic cells after the in vitro X-irradiation was measured by flow cytometry after incubation periods of 24, 48 and 72 hours. We also observed the apoptotic cells using a DNA fragmentation assay and electron microscopy. RESULTS: The rate of spontaneous apoptosis increased in relation to the time interval following irradiation (1.761+/-0.161, 3.563+/-0.564, 11.098+/-2.849, at 24, 48, and 72 hours). The apoptotic cells also increased in the samples irradiated with 0.5, 1, 2 and 5 Gy, in a radiation dose and time interval after irradiation manner, with the apoptosis being too great at 72 hours after irradiation. The dose-response curves were characterized by an initial steep increase in the number of apoptotic cells for irradiation doses below 2 Gy, with a flattening of the curves as the dose approached towards 5 Gy. CONCLUSION: The flow cytometric assay technique yielded adequate data, and required less than 1 mL of blood. The time and dose dependence of the radiation-induced apoptosis, was also shown. It is suggested that the adequate time interval required for the evaluation of apoptosis would be 24 to 48 hours after blood sampling.

Mantle Cell Lymphoma, Blastoid Variant, Diagnosed on the Basis of Cytomorphology and Flow Cytometric Immunophenotyping of the Lymph Node Aspirate and Peripheral Blood

Mantle cell lymphoma, blastoid variant (B-MCL), is a very rare type of non-Hodgkin 's lymphoma exhibiting an aggressive clinical course. We describe a case of B-MCL showing generalized lymphadenopathy and leukemic conversion in a 62-yr-old man. The case was diagnosed and subclassified as B-MCL on the basis of cyto-morphology and immunophenotype. Microscopic examination of the peripheral blood (PB) showed a spectrum of cells ranging from small mature lymphocytes to medium- and large-sized lymphocytes with blast-like chromatin and prominent nucleoli. The lymphoma cells were monoclonal B cells with moderately intense surface IgM. They were CD5 positive, cyclin D1 positive, CD10 negative, and CD23 negative. The flow cytometric immunophenotyping and DNA ploidy analysis of the PB and material obtained by aspiration cytology supported the diagnosis of B-MCL. These findings underline the utility of aspiration cytology in diagnosing B-MCL when cytomorphologic examination is combined with flow cytometric analysis of immuno-phenotype and demonstration of proliferation markers.

Clinical Correlation of iNOS and p53 Expression in Colorectal Cancer

PURPOSE: Nitric oxide (NO) is synthesized from the amino acid L-arginine by nitric oxide synthase (NOS) which exists as three isoforms, the calcium-dependent endothelial NOS and neuronal NOS, and a calcium-independent inducible NOS. NO has been studied in a variety of human cancers and is implicated in both tumor promotion and inhibition. NO cause p53 mutation in human cells and mutations of p53 are the most common genetic abnormality yet found in human cancers. Aims of this study is to investigate the correlation of iNOS and p53 expression in colorectal cancer, and to evaluate its clinicopathological siginificance with the expression of these proteins. METHODS: 125 patients, who received curative resection of colorectal cancer from 1992 to 1996, were analyzed retrospectively. The monoclonal antibody to the iNOS (Transduction Laboratories.), the monoclonal antibody to the mutant p53 (Dako Co.) were used for the immunohistochemical analysis. Normal colorectal tissue were assayed in 45 cases. The relationship between mutant p53 and iNOS was investigated. RESULTS: When iNOS expression were detected in specimens, positive rate of mutant p53 were 72.4%. When mutant p53 expression were detected in specimens, positive rate of iNOS expression were 65.8%(P<0.05). Positive rate of iNOS, mutant p53 in tumor size were 51%, 56% below 4 cm and 60%, 63% in 4~8 cm and 47%, 67% above 8 cm. Positive rate of iNOS, mutant p53 in Dukes' stage were55%, 55% in stage B and 56%, 67% in stage C. Positive rate of iNOS, mutant p53 in histologic differentiation were 55%, 55% in well-differentiation and 61%, 66% in moderate differentiation and 35%, 48% in poor-differentiation. There was no difference in each Dukes stage between iNOS expression or p53 mutation and postop five year survival rate. Positive rate of iNOS, mutant p53 in normal tissue were 22%, 32%. CONCLUSION: The prevalence of iNOS expression and p53 mutation has been found in exceeding 50% of cases. There was a significant correlation between iNOS expression and p53 mutation in colorectal cancer. No correlation was found between iNOS expression or p53 mutation and clinicopathologic parameters.

Lymphopenia after Mediastinal Irradiation in Lung Cancer / 대한방사선종양학회지

PURPOSE: This study was undertaken to retrospectively evaluate white blood cell kinetics, especially lymphocyte depression after different treatments, and to find the correlation between immunosuppression and large blood volume and dynamic blood flow within the mediastinal radiotherapy (RT) field in lung cancer. MATERIALS AND METHODS: Thirty-four patients with lung cancer were retrospectively evaluated; 10 patients had only radiotherapy (RT group), 8 had chemotherapy (CT group) and 16 had chemotherapy and radiotherapy (RT/CT group). The mean follow-up periods of the RT-including groups (RT group and RT/CT group) and the RT-excluding group (CT group) were 6 and 8 months, respectively. Complete blood cell counts including lymphocyte percentage (%) were checked weekly during RT but less frequently during CT and after RT. RESULTS: Changes in total white blood cell counts were not significantly different among the three groups. The lymphocyte count and lymphocyte % were much lower in the RT-including groups than in the RT- excluding group. The difference between pre-treatment and final lymphocyte count and the difference between pre-treatment and final lymphocyte % were significant (p=0.044 and p=0.037) between the RT- including groups and the RT-excluding group. CONCLUSION: Lymphopenia was more marked after treatment containing RT than CT only. Lymphopenia may be one cause of a compromised immune system after mediastinal irradiation in lung cancer. We suggest cautiously that previous studies showing evidence of lymphocyte apoptosis after low-dose irradiation and large blood volume and dynamic blood flow within the RT fields could be somewhat related to lymphopenia after mediastinal irradiation.

Adenoid Basal Carcinoma of the Uterine Cervix

We report a case of adenoid basal carcinoma of the uterine cervix unexpectedly found in a 68-year-old female. She was diagnosed with a high-grade squamous intraepithelial lesion (HSIL) on a cervical smear and subsequent cervical punch biopsy. Total abdominal hysterectomy was performed, and there were no significant gross findings in the uterine cervix. Microscopically, the epithelial surface showed a HSIL with glandular extension. Below the neoplastic epithelial lesion were numerous small nests of uniform small cells, which extended to the parametrium. These tumor cells contained hyperchromatic nuclei, inconspicuous nucleoli, and scant cytoplasm. Peripheral palisading and gland-like or acinar structures in the nests were noted. The latter were positive for mucicarmine stain. Foci of squamous differentiation exhibiting occasional mitoses and large atypical cells were seen in the small nests. Stromal reaction was not obvious. Immunohistochemically, the HSIL lesion and adenoid basal carcinoma lesion were negative for human papillowa virus. The tumor cells forming nests were positive for carcinoembryonic antigen, S-100 protein, and high molecular weight cytokeratin, but were negative for -smooth muscle actin and chromogranin A.

Study on Immunohistochemical Expression of p53, Epidermal Growth Factor and c-erbB-2 in Squamous Cell Carcinoma of the Head and Neck / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: Cell proliferation and differentiation are regulated by growth factors and growth factor receptors. Inappropriate expressions of growth factors and oncogenes may influence the biological and clinical phenotype of tumor. But there are still controversy regarding their clinical attributions in head and neck cancer. The authors evaluated the prognostic significance of p53 protein, EGF and c-erbB-2 expression in the squamous cell carcinoma of head and neck to determine their relationship with the clinicopathologic parameters. MATERIALS AND METHODS: Paraffin embedded tissue specimen from 64 cases of squamous cell carcinomas of head and neck were studied by immunohistochemical staining. RESULT: p53 protein, EGF and c-erbB-2 were expressed 53.1%, 29.7% and 57.8%, respectively, in 64 cases of head and neck cancer. The positive expression of p53 protein was associated closely with T-stage and clinical stage. This expression was statistically significant(P<0.05). There were no significant relationships between the reactivity of EGF, c-erbB-2 and clinical parameters. CONCLUSION: The expressions of p53 protein, EGF and c-erbB-2 could be related to oncogenesis in squamous cell carcinoma of the head and neck. And, the expression of p53 protein can be used as a prognostic factor in head and neck squamous cell carcinoma.

Ameloblastic fibro-odontoma: a case report

Metastatic Cervical Lymphadenopathy from Uterine Leiomyosarcoma with Good Local Response to Radiotherapy and Chemotherapy / 대한방사선종양학회지

The metastasis of uterine leiomyosarcoma to the neck node has not been reported previously and the radiotherapy has been rarely used for the metastatic lesion of the other sites. We report a case of neck metastasis from a uterine leiomyosarcoma, which developed 10 months after surgery and postoperative pelvic radiotherapy. It also involved the parapharyngeal space, adjacent spine, and spinal canal. The metastatic neck mass was inoperable, and was treated by neck radiotherapy (6,000 cGy) and chemotherapy including taxol and carboplatin. The mass has regressed progressively to a nearly impalpable state. She has never developed spinal cord compression syndrome, and has maintained good swallowing for eight months since the neck radiotherapy and chemotherapy. Since the extensive metastatic neck mass showed good local response to high dose radiotherapy and chemotherapy, both treatments may be considered for an unresectable metastatic leiomyosarcoma.

Expression of c-erbB-2, c-myc, c-fos, bcl-2, p53, PCNA, and TGF-alpha in Transitional Cell Carcinoma of the Urinary Bladder

Most of malignant tumors in the urinary bladder is transitional cell carcinoma (TCC) deriving from the urothelium. Clinical stage and histopathologic grading of the TCC of the urinary bladder is important in the determination of the patient's prognosis. To investigate the correlation between the prognostic factors and the expression of the various oncoproteins and growth factors in each grade of the TCC, immunohistochemical stains for c-erbB2, c-myc, c-fos, bcl-2, p53, proliferating cell nuclear antigen (PCNA), and transforming growth factor-alpha (TGF-alpha) were performed in the formalin fixed paraffin embedded tissues of the TCC (Grade I; 15 cases, Grade II; 20 cases, Grade III; 15 cases) of the urinary bladder. The immunoexpression rate of c-erbB2 was immunoexpression 78.0% in the grade I, 85.0% in the grade II, and 95.0% in the grade III TCC. The immunoexpression rate of c-myc, c-fos and bcl-2 was below 5% in each grades of TCC. The p53 immunoexpression was identified in 11.5%, 24.3% and 30.6% of the grade I, II, and III TCC, respectively. The PCNA and TGF-alpha expression was 53.0% and 27.6% in the grade I, 77.3% and 32.7% in the grade II, and 78.2% and 37.3% in the grade III TCC, respectively. These results suggest that the expressions of c-myc, c-fos, bcl-2, and TGF-alpha are similar in each grade of the TCC and the positivity of c-erbB2, p53, and PCNA shows an increasing tendency for the higher grade TCC of the urinary bladder. Therefore, c-erbB2, p53, and PCNA are clinically useful predictors of the patient's prognosis.

Relationship of HPV Detection and p53 Expression in Urinary Bladder Cancer / 대한암학회지

PURPOSE: The purpose of this study was to investigate the relationship between aberrant p53 expression and the presence of human papillomavirus DNA in transitional cell carcinomas of the urinary bladder. MATERIALS AND METHODS: This study analyzed 30 paraffin-embedded transitional cell carcinoma of the urinary bladder, including 10 cases of grade I, 10 cases of grade II, and 10 cases of grade III, for the presence of DNA-HPV and abnormal accumulation of p53. We used immunohistochemical staining for p53 protein and in situ hybridization for HPV DNA, respectively. RESULTS: Overall positive rate of HPV DNA type 16 and type 18 was 60.0% and 53.3%, respectively. Nuclear accumulation of p53 was found in 13 cases (43.3%) of all transitional cell carcinomas. In HPV DNA type 16 positive cases, the p53 was positive in 8 cases and negative in 10 cases. In HPV DNA type 16 negative cases, the p53 was positive in 5 cases and negative in 7 cases. In HPV DNA type 18 positive cases, the p53 was positive in 7 cases and negative in 9 cases. In HPV DNA type 16 negative cases, the p53 was positive in 6 cases and negative in 8 cases. CONCLUSION: This results suggest that HPV infection and p53 gene accumulation may contribute to the significant role in the carcinogenesis of the utinary bladder. In additon, HPV infection and p53 accumulation may be related to tumor progression and higher grade.