Atsushi Hongo

Vascular endothelial growth factor-C expression and its relationship to pelvic lymph node status in invasive cervical cancer.

Vascular endothelial growth factor-C (VEGF-C) has been implicated in lymphangiogenesis, the process of new lymphatics formation. The present study investigated VEGF-C mRNA expression in invasive cervical cancer tissue. Additionally, the association of VEGF-C mRNA with clinicopathological features was examined. VEGF-C mRNA expression was assessed by reverse transcription-polymerase chain reaction using β-action as an internal control. 75 patients presenting with invasive cervical cancer were included in the trial. VEGF-C mRNA expression was markedly higher in tumours in which pelvic lymph node metastasis was diagnosed by magnetic resonance (MR) imaging (P = 0.002). 53 patients displaying stage Ib–IIb cervical cancer underwent radical hysterectomy and pelvic lymphadenectomy. VEGF-C expression was significantly higher in tumours exhibiting deep stromal invasion, pelvic lymph node metastasis and lymph-vascular space involvement (P = 0.016, P = 0.006 and P = 0.036, respectively). Multivariate analysis revealed VEGF-C mRNA expression to be the sole independent factor influencing pelvic lymph node metastasis. Subjects demonstrating VEGF-C mRNA expression displayed significantly poorer prognoses than those lacking VEGF-C mRNA expression (P = 0.049). These findings provide evidence supporting the involvement of VEGF-C expression in the promotion of lymph node metastasis in cervical cancer. Furthermore, examination of VEGF-C expression in biopsy specimens may be beneficial in the prediction of pelvic lymph node metastasis.

Rapid and sensitive detection of physical status of human papillomavirus type 16 DNA by quantitative real-time PCR

ABSTRACT A rapid quantitative real-time PCR method was employed to quantify the copy number of E2 and E6 genes for analysis of the physical status of human papillomavirus type 16 (HPV-16) DNA. Significant differences with respect to both copy numbers were found when more than 40% of HPV-16 DNA was integrated with disruption of the E2 gene in an experimental model. The physical status of HPV-16 DNA in 50 clinical samples was exclusively episomal in 21 cases (42%), concomitant in 14 cases (28%), and integrated in 15 cases (30%). The prevalence of integrated and/or concomitant forms of HPV-16 DNA increased with progression of cervical disease. Four of 11 cervical intraepithelial neoplasia involved integrated forms of HPV-16 DNA partially or exclusively. This rapid, sensitive technique is useful in the analysis of the physical status of HPV DNA.

Vascular endothelial growth factor is implicated in early invasion in cervical cancer.

The association between the expression of vascular endothelial growth factor (VEGF) and clinicopathological factors has scarcely been examined in cervical cancer. This study examines the level of VEGF messenger RNA (mRNA) expression in invasive cervical cancer and its association with clinicopathological features including microvessel density. The level of VEGF mRNA was assessed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) using beta-actin as an internal control in 66 patients with stages Ia-IVb invasive cervical cancer. In 42 patients who underwent surgery, the microvessel count was also assessed by immunostaining for factor VIII-related antigen in the most neovascularised area of the specimen. The highest level of VEGF mRNA expression was observed in early invasive cervical cancers. Except for stage IVb, the stage of the disease inversely correlated with the level of VEGF mRNA (P < 0.05). There was no significant difference in the level of VEGF mRNA with respect to histological cell types. 38 patients with stages Ib-IIb cervical cancer underwent radical hysterectomy and pelvic lymphadenectomy. There was no significant difference in the level of VEGF mRNA with respect to lymph node metastasis, depth of stromal invasion, tumour size, parametrial involvement or vaginal involvement among these patients. A significant relationship was found between the microvessel density and the level of VEGF mRNA (P < 0.01). These findings provide evidence that the expression of VEGF is involved in the promotion of angiogenesis in cervical cancer and plays an important role in early invasion.

Factors associated with parametrial involvement in stage IB1 cervical cancer and identification of patients suitable for less radical surgery

OBJECTIVE The purpose of the present study was to determine possible factors associated with parametrial spread in patients with stage IB1 cervical cancer and define parameters associated with a low risk for parametrial spread, in order to identify candidates for less radical surgery. PATIENTS AND METHODS We retrospectively reviewed 200 patients with stage IB1 cervical cancer who had undergone radical hysterectomy (class III) and pelvic lymphadenectomy. RESULTS Overall, 20 (10.0%) of the 200 patients revealed parametrial spread, of which 11 (55%) had only direct microscopic extension of the disease, 3 (15%) had only disease spread to parametrial lymph nodes, 1 (5%) had both direct microscopic extension and disease spread to parametrial lymph nodes, and 5 (25%) had only tumor emboli within the lymph vascular channels in the parametrial tissue. Elderly age, depth of invasion, tumor size, lymph vascular space invasion (LVSI), positive pelvic nodes, and ovarian metastasis were significantly associated with parametrial involvement. The multivariate analysis model included factors that could be determined by a cone biopsy and showed LVSI, deep stromal invasion, and elderly age to be the independent predictors of parametrial involvement. Ninety-one patients had a depth of invasion of ≤10 mm and no LVSI, of which only 1 (1.1%) had parametrial involvement. When patients aged ≤50 years were further stratified into those with a depth of invasion of ≤10 mm and no LVSI, parametrial involvement was found to be 0.0% (0/68). CONCLUSION Patients with a tumor depth of invasion of ≤10 mm, no LVSI, and aged ≤50 years, could be considered for less radical surgery such as modified radical hysterectomy or simple hysterectomy with pelvic lymphadenectomy.

The SUVmax of 18F-FDG PET correlates with histological grade in endometrial cancer.

Objectives: The objectives of this study were to assess the maximum standardized uptake value (SUVmax) of 18F-fluorodeoxyglucose (18F-FDG) by a primary tumor of endometrial cancer with the use of positron emission tomography/computed tomography (PET/CT) and to assess its association with the clinical importance of the disease. Methods: 18F-fluorodeoxyglucose PET/CT scan was performed on 44 participants within 2 weeks before surgery. 18F-fluorodeoxyglucose uptake was quantified by calculating the SUVmax. The distribution of cases that scored positive for each of the biological parameters examined was correlated with the SUVmax of the 18F-FDG PET/CT and the glucose transporter-1 expression status obtained by immunohistochemistry. Results: The mean SUVmax of the primary endometrial cancer tumors was 17.6 (range, 3.04-34.74). There were significant correlations between the SUVmax of the primary tumor and the International Federation of Gynecology and Obstetrics (FIGO) grade (P < 0.001), maximum tumor size (P < 0.001), and glucose transporter-1 expression (P < 0.001). Furthermore, multivariate analysis showed that the FIGO grade was most significantly identified as a relation factor of SUVmax (≥17.6) for endometrial cancer (P = 0.017). The present findings indicate that a significant relationship was seen between the SUVmax and the FIGO grade in endometrial cancer. Conclusion: We propose that the primary tumors SUVmax obtained from 18F-FDG PET/CT may be associated with aggressive biological characteristics in endometrial cancer.

Measurement of the minimum apparent diffusion coefficient (ADCmin) of the primary tumor and CA125 are predictive of disease recurrence for patients with endometrial cancer.

OBJECTIVE The purpose of this study was to evaluate whether preoperative measurements of the minimum apparent diffusion coefficient (ADCmin) on magnetic resonance imaging (MRI) and the tumor marker CA125 are correlated with the clinical characteristics and prognosis of patients with endometrial cancer. METHODS The distribution of cases that scored positive for each of the biological parameters examined and the correlations with the ADCmin of the primary tumor and the serum tumor marker CA125 were examined for 111 patients with preoperative assessment of primary endometrial cancer. RESULTS There were significant correlations between the ADCmin of the primary tumor and the FIGO stage (P=0.001), depth of myometrial invasion (P<0.001), cervical involvement (P=0.003), lymph node metastasis (P=0.027), ovarian metastasis (P<0.001), peritoneal cytology (P=0.027) and tumor maximum size (P<0.001). The disease-free survival (DFS) rate of patients with high serum CA125 was significantly lower than that of patients with low serum CA125 (P=0.0395). The DFS rate of patients with a low ADCmin of the primary tumor was significantly lower than that of patients with a high ADCmin of the primary tumor (P<0.001). In particular, the ADCmin of the primary tumor was an independent factor for disease recurrence in a multivariate analysis (P=0.019). CONCLUSIONS The present findings indicate that a low preoperative ADCmin of the primary tumor is an important predictive factor for identifying endometrial cancer patients with a risk of disease recurrence.

The measurement of SUVmax of the primary tumor is predictive of prognosis for patients with endometrial cancer

OBJECTIVE The purpose of this study was to evaluate if preoperative measurements of the maximum standardized uptake valve (SUVmax) on positron emission tomography/computed tomography (PET/CT) and tumor marker CA125 are correlated with clinical characteristics and prognosis in patients with endometrial cancer. METHODS The distribution of cases that scored positive for each of the biological parameters examined and the correlations with the SUVmax of the primary tumor and the serum tumor marker CA125 were examined for 106 patients with preoperative assessment of primary endometrial cancer. RESULTS There were significant correlations between the SUVmax of the primary tumor and the FIGO stage (P=0.030), histology (P=0.025), depth of myometrial invasion (P=0.031) and tumor maximum size (P<0.001). The serum CA125 level was significantly associated with the FIGO stage (P=0.050). The disease-free survival (DFS) and overall survival (OS) rates of patients exhibiting a high SUVmax of the primary tumor were significantly lower than those of patients exhibiting a low SUVmax of the primary tumor (P=0.049, and P=0.039, respectively). Furthermore, the DFS and OS rates of patients exhibiting a high SUVmax of the primary tumor were significantly lower than those of patients exhibiting a low SUVmax of the primary tumor at advanced stages (stages III-IV) (P=0.032 and P=0.023, respectively). In particular, the SUVmax of the primary tumor was an independent prognostic factor for OS by a multivariate analysis (P=0.025). CONCLUSIONS The present findings indicate that for patients with endometrial cancer, a high preoperative SUVmax of the primary tumor is an important predictive factor for identifying endometrial cancer patients with a poor prognosis.

The mean apparent diffusion coefficient value (ADCmean) on primary cervical cancer is a predictive marker for disease recurrence

OBJECTIVE The purpose of this study is to investigate the correlation of the max, mean and minimal apparent diffusion coefficient values (ADCmax, ADCmean, and ADCmin) on diffusion weighted imaging findings with prognostic factors in cervical cancer. METHODS A cohort of 80 cervical cancer patients underwent pelvic magnetic resonance imaging (MRI) within the 2 to 4 weeks prior to radical hysterectomy. The optimal cutoff value for segregating disease free survival (DFS) was determined by receiver operating characteristic (ROC) curve analysis. We used ROC curve analyses to evaluate whether preoperative ADCmax, ADCmean, ADCmin on MRI predicted the risk group of recurrence. RESULTS Analyses of ROC curves identified an optimal The ROC curves identified an optimal ADCmax, ADCmean, and ADCmin cutoff values of 1.122 × 10(-3)mm(2)/s, 0.852 × 10(-3)mm(2)/s, 0.670 × 10(-3)mm(2)/s and for predicting the recurrence of cervical cancer. The patients categorized into the lower ADCmean or ADCmin groups showed the shorter disease free survivals compared with the higher ADCmean or ADCmin, respectively (P<0.0001 or P=0.0210). In particular, the ADCmean of primary cervical cancer was an independent predictive factor for disease recurrence by a multivariate analysis (P=0.0133). CONCLUSIONS The ADCmean of primary cervical cancer calculated by MRI could be an important factor for identifying patients with a risk of disease recurrence.

Vascular endothelial growth factor and platelet-derived endothelial cell growth factor expression are implicated in the angiogenesis of endometrial cancer.

Although many angiogenic factors have been described, it is not well defined which factors are expressed in endometrial cancer. The object of this study was to examine mRNA levels of the two angiogenic factors, vascular endothelial growth factor (VEGF) and platelet-derived endothelial cell growth factor (PD-ECGF) in endometrial cancer tissues and their association with clinicopathological features including microvessel density. The level of VEGF and PD-ECGF mRNAs was assessed by semi-quantitative reverse transcription-polymerase chain reaction using beta-actin as an internal standard in 38 patients with endometrial cancer. Microvessel counts were also assessed by immunostaining for factor VIII-related antigen in the most vascularised area of the specimen. VEGF/beta-actin ratios of non-endometrioid tumours were significantly higher than those of endometrioid tumours (P = 0.013). VEGF/beta-actin ratios of cases with lymph-vascular space involvement were significantly higher than those of cases without lymph-vascular space involvement (P = 0.021). Although it was not statistically significant, PD-ECGF/beta-actin ratios in grade 3 tumours were higher than those in grade 1 and 2 tumours (P = 0.066). The microvessel density was significantly correlated with the level of VEGF and PD-ECGF mRNA expression (P = 0.041 and P < 0.0001, respectively). Our findings provide evidence that the expression of both VEGF and PD-ECGF is involved in the promotion of angiogenesis in endometrial cancer. In addition, VEGF and PD-ECGF might contribute to the aggressive potential of high grade tumours or certain histological subtypes with unfavourable prognosis through the induction of angiogenesis.

IGFs and Cell Growth

A tissue or an organ grows when the number of cells produced per unit time exceeds the number of cells that die in the same period. This rather obvious statement often has been obscured by the fact that most investigators use cells in culture for basic studies. Such a use has innumerable advantages, but, on the other hand, under optimal conditions, cells in culture grow exponentially, with very little cell death, which is therefore largely neglected. However, in animals, physiological cell death occurs constantly, so that the size of any population of cells, whether normal or abnormal, depends on both the rate of cell division and the rate of cell death (1). More precisely, an increase in cell number of any given cell population depends on three parameters: The first is the length of the cell cycle. A shortening of the cell cycle will cause the cells to divide more frequently, resulting in an increase in cell number. The second is the growth fraction (2), i.e., the fraction of cells in a population that are actively dividing in the cell cycle. In many cell populations, a sizable fraction of cells is in GO, a distinct phase of the cell cycle, in which the cells are quiescent, and do not traverse the cell cycle, although they are alive and capable, under certain conditions, of responding to proliferative stimuli. An increase in the growth fraction, i.e., a decrease in the GO fraction, will also result in an increased production of cells. The third is a decrease in the rate of cell death. The third group was all but ignored until recently, when apoptosis suddenly became the fashionable way for cells to die. But it had been known for several years that the rate of cell proliferation (the first and second parameters) was not sufficient to explain why tumors grow and normal tissues in the adult animal do not. Baserga and Kisieleski (3) were the first to show that the cells of one of the fastest growing tumors of mice did not proliferate faster than the normal cells of the lining epithelium of the small intestine. The obvious explanation was that the epithelial cells of the intestinal mucosa constantly undergo renewal, with a cell dying at the tip of the villi for every cell generated in the crypts. In other words, the size of the population of the very actively dividing epithelial cells of the intestinal mucosa does not increase, because the number of cells produced per unit time equals the number of cells that die in the same time period. Later, Steel et al. (4) convincingly showed that, in tumor growth, some extent of cell death had to be postulated. Finally, Bresciani et al. (5) showed that, in human squamous cell carcinomas of the skin, tumor cells also die in large amounts, but the tumors grew because the rate of cell division always exceeded the rate of cell death.