Robotic Vaginal Cuff Closure During Radical Hysterectomy for Early-Stage Cervical Cancer: The Bruges Method.

The only randomized trial (LACC trial, Laparoscopic Approach to Cervical Cancer), published in 2018, comparing the oncologic outcomes of minimally invasive and open surgery in early-stage cervical cancer, has shown inferior disease-free and overall survival for minimally invasive surgery. Subsequent large retrospective cohort studies of centers with long-standing experience in minimally invasive surgery and large nationwide cohort studies have shown that both the laparoscopic and robotic approaches have similar survival outcomes as the open surgery group in the LACC trial. Important protective measures to avoid tumor spillage in the peritoneal cavity during colpotomy were the closure of the vaginal cuff and avoiding the use of a uterine manipulator. Several methods have been described to close the vaginal cuff, mainly by a vaginal approach. Here we describe with a video a new technique of vaginal cuff closure during a robotic-assisted radical hysterectomy. During the robotic procedure, a purse string barbed suture is placed through the vaginal walls in order to close the vagina prior to colpotomy. The technique is a feasible, relatively fast, and easy-to-learn addition to the robotic radical hysterectomy procedure in early-stage cervical cancer.

Robotic para-aortic sentinel lymph node mapping in endometrial, cervical, and ovarian cancer.

The concept of pelvic sentinel lymph node mapping has been well-investigated in endometrial and cervical cancer. A variety of tracers have been used including blue dye, technetium-99-m (Tc-99 m), and fluorescent tracer indocyanine green. Pelvic sentinel lymph node mapping has shown its safety, efficacy, and diagnostic accuracy, with high sensitivity and negative predictive value of more than 90%, in retrospective cohort studies as well as in prospective trials for robotic surgery. The concept of pelvic sentinel lymph node biopsy has been incorporated in several international guidelines in early-stage endometrial cancer and a subgroup of early-stage cervical cancer, although survival data are still needed to confirm its standard use. The application of para-aortic sentinel lymph node mapping is still in a development phase, but its detection rate and diagnostic accuracy seem to be promising in initial studies. Here, an overview is given of the recent developments in the different methodologies used for identifying para-aortic sentinel lymph nodes in endometrial, cervical, and ovarian cancer.

Primary leiomyosarcoma of the uterine cervix: an unusual case and critical appraisal.

Leiomyosarcomas of the uterine cervix are rare, mostly occurring in perimenopausal women. Diagnosis is based on pathology and immunohistochemistry. Surgery with a total abdominal hysterectomy and bilateral salpingo-oophorectomy remains the standard. A female patient in her 60s presented with heavy postmenopausal bleeding. Vaginal ultrasound scan and magnetic resonance imaging showed a large strongly vascularized cervical mass with features suspicious of sarcomatous degeneration. Positron Emission Tomography-Computed Tomography (PET-CT) did not reveal any evidence of metastases nor lymphadenopathy, but presence of right hydronephrosis. An abdominal hysterectomy with bilateral salpingo-oophorectomy, and end-to-end anastomosis of the right ureter, was performed. Pathology showed an International Federation of Gynecology and Obstetrics (FIGO)-stage 1B leiomyosarcoma of the uterine cervix. No adjuvant treatment was given. Adjuvant radiotherapy reduces the risk of recurrence but no survival impact. The benefit of adjuvant chemotherapy is questionable given the lack of randomized trials. Multidisciplinary research concerning molecular alterations of the disease is required to determine optimal management strategies with potential novel molecular therapies.

Case report: a robotic-vaginal approach for total vaginectomy and hysterectomy with pelvic sentinel lymph node dissection in primary vaginal melanoma: a 10-step technique and literature review.

Introduction: Primary vaginal melanoma is extremely rare, has a poor prognosis, and occurs mostly in elderly women. The diagnosis is based on histology and immunohistochemistry of a biopsy. Given the rarity of vaginal melanoma, no standardized treatment guidelines are established; however, surgery is the primary treatment modality in the absence of metastatic disease. Most reports in the literature are retrospective single cases, case series, and population-based studies. The open surgical approach is the main modality reported. Here, we report for the first time a 10-step combined robotic-vaginal technique, with en bloc resection of the uterus and total vagina, for treating clinically early-stage primary vaginal melanoma. In addition, the patient in our case underwent a robotic pelvic bilateral sentinel lymph node dissection. The literature on the surgical approach for vaginal melanoma is reviewed. Case presentation: A 73-year-old woman was referred to our tertiary cancer center and was clinically staged according to the 2009 International Federation of Gynaecology and Obstetrics (FIGO) staging for vaginal cancer as FIGO-stage I (cT1bN0M0) and according to the American Joint Committee on Cancer (AJCC) for (cutaneous) Melanoma Staging as clinical stage IB. Preoperative imaging with magnetic resonance imaging, FDG-positron emission tomography-computed tomography, and ultrasound of the groins did not reveal any adenopathy nor metastases. The patient was planned for a combined vaginal and robotic en bloc total vaginectomy and hysterectomy, as well as a pelvic bilateral sentinel lymph node dissection. Results: The surgical procedure was performed in 10 steps described in this case report. The pathology revealed free surgical margins and negative test results for all sentinel lymph nodes. The postoperative recovery process was uneventful, and the patient was discharged on day 5. Conclusion: The main surgical approach reported for primary early-stage vaginal melanoma is open surgery. A minimally invasive surgical approach, described here as a combined vaginal-robotic en bloc total vaginectomy and hysterectomy, for the surgical treatment of early-stage vaginal melanoma enables precise dissection, low surgical morbidity, and fast recovery for the patient.

Robotic surgery in early and advanced ovarian cancer: Case selection for surgical staging and interval debulking surgery.

OBJECTIVE: During the last decade several case series have been published on robotic surgery in early and advanced stage ovarian cancer. Although most studies lack a significant oncological follow-up, more importantly criteria for patient selection for both robotic surgical staging (R-SS) and robotic interval debulking surgery (R-IDS) are not well defined. The objective of this study was to assess the surgical and oncological outcomes, using well-defined selection criteria, between robotic and open surgery in early and advanced stage ovarian cancer. STUDY DESIGN: Single-center retrospective case cohort study including 96 ovarian cancer patients. For early stage ovarian cancer, patients were selected for R-SS after laparoscopic salpingo-oophorectomy of a suspicious adnexal mass. For advanced stage ovarian cancer, only patients receiving neoadjuvant chemotherapy and IDS were included in the study. Exclusion criteria were the presence of residual peritoneal disease after NACT and/or patients requiring additional complex surgical procedures. RESULTS: For early stage ovarian cancer, similar median operative times were seen between R-SS and open surgical staging (O-SS), 132 min and 120 min respectively. Pelvic/para-aortic lymph node yield was similar between R-SS and O-SS, 22/11 nodes and 18/8 nodes respectively. Surgical upstaging occurred in 11.5% in the R-SS group and in 27.6% in the O-SS group. In advanced stage ovarian cancer, the BMI was significantly higher in the R-IDS group compared to the O-IDS group (27.8 vs 23.5; p =.006). The median follow was 52 months in the R-IDS group and 31 months in the O-IDS group. Recurrent disease occurred in 42.9% of the R-IDS group and in 45% of the O-IDS group. The length of hospitalization was significantly longer in the O-SS and O-IDS group (p <.00001). CONCLUSION: Patients with clinically early stage ovarian cancer, confirmed after laparoscopic removal of a suspicious adnexal mass, are candidates for R-SS whilst maintaining similar surgical and oncological outcome measures as O-SS. In advanced ovarian cancer, suitable candidates for R-IDS are those who receive NACT with good response and no residual peritoneal disease, especially in patients with a high BMI, but large prospective randomized trials with well-defined criteria are needed.

Robotic Staging of Cervical Cancer With Simultaneous Detection of Primary Pelvic and Secondary Para-Aortic Sentinel Lymph Nodes: Reproducibility in a First Case Series.

Objective: Discrepancies exist among international guidelines on the surgical staging of para-aortic lymph nodes in locally advanced cervical cancer (LACC), varying from considering a para-aortic lymph node dissection, at least up to the inferior mesenteric artery, to a complete para-aortic lymph node dissection. In this study, we aim to assess the reproducibility of our recently reported robotic technique using indocyanine green for identifying besides primary pelvic sentinel lymph nodes (SLNs), secondary para-aortic SLNs in a first case-cohort of cervical cancer patients. Methods: A retrospective case series of LACC patients with/without suspicious pelvic lymph nodes (LNs) on imaging (including two patients with an additional suspicious para-aortic LN) is reported. All patients underwent a robotic pelvic SLN and para-aortic sentinel/nonsentinel LN dissection using the da Vinci Xi platform. Indocyanine green was used as a fluorescent tracer, at a concentration of 1.9 mg/mL, and injected as 0.5 mL in each quadrant of the cervix. Results: In a total of 10 cases, primary pelvic SLNs (90% bilateral) with subsequent secondary para-aortic SLNs were identified in all cases. Lower para-aortic SLNs were present in all cases, and upper para-aortic SLNs were found in 9 out of 10 cases. The mean age of the cervical cancer patients was 49.8 years (SD ± 6.89), and the mean body mass index (BMI; kg/m2) was 23.96 (SD ± 4.60). The median total operative time was 105.5 min (range: 89-141 min). The mean numbers of primary pelvic SLNs and secondary lower and upper para-aortic SLNs were 3.10 (SD ± 1.10), 2.90 (SD ± 0.74), and 2.30 (SD ± 1.57), respectively. The median number of total para-aortic LNs (PALNs) dissected per patient was 11.5. Six patients had positive primary pelvic SLNs, and two had secondary positive para-aortic SLNs. The nonsentinel para-aortic LNs were negative in all cases. There were no intra- or postoperative complications. Conclusion: Our preliminary experience demonstrates the reproducibility of identifying, besides primary pelvic SLNs, secondary lower and upper para-aortic SLNs during robotic staging in LACC. A surgical approach limiting a complete para-aortic LN dissection could reduce the potential risks and morbidity associated with this procedure. To determine the sensitivity and negative predictive value of this new surgical approach, and whether the lower para-aortic SLNs under the inferior mesenteric artery are representative of the whole para-aortic region, large prospective observational studies are needed in LACC and/or those with suspicious pelvic LNs but apparent normal para-aortic LNs on imaging.

Indocyanine green guidance for combined robotic pelvic and para-aortic sentinel node mapping in cervical cancer.

BACKGROUND: Indocyanine green (ICG) for pelvic sentinel lymph node (SLN) mapping is well established in endometrial cancer (Persson et al., 2019 Jul). However, the application for para-aortic SLNs is less reported; and the detection rate of para-aortic SLNs, mainly after cervical injection of ICG, varies between 14% and 71% (Rossi et al., 2013 Nov; Kim et al., 2020 Mar; Gallotta et al., 2019 Mar). One recent report differentiates between lower and upper para-aortic SLNs in endometrial cancer (Kim et al., 2020 Mar). Here we describe a technique using ICG for identifying pelvic SLNs, lower and upper para-aortic SLNs in cervical cancer. VIDEO: A 46-year old female presented with high grade cervical dysplasia/carcinoma in situ on cervical smear. Cervical cone biopsy revealed a grade two squamous cell carcinoma (depth of invasion 6.8mm, width 20.8mm). Clinically she was staged as an early FIGO-stage IB2 cervical cancer. NMR revealed bilaterally enlarged iliac lymph nodes. Additional PET-CT revealed FDG-uptake in the enlarged pelvic lymph nodes. In view of the imaging findings a staging Robotic pelvic and para-aortic SLN procedure was planned, prior to select the primary treatment (radical hysterectomy or chemo-radiation). ICG was injected into the cervical stroma, and a robotic pelvic and para-aortic SLN dissection (using Firefly System ®, Intuitive Surgical Inc.) was initiated 15 minutes and 35 minutes, respectively, after cervical injection. RESULTS: This video demonstrates the application of ICG for mapping bilateral primary pelvic SLNs, secondary and tertiary para-aortic SLNs in the lower and upper para-aortic region respectively, in cervical cancer. Pathology revealed one metastatic pelvic SLN on the left side, other four pelvic SLNs were negative; both the secondary/lower (n = 3) and tertiary/upper (n = 5) para-aortic SLNs were negative, as well as the non-SLNs (n = 8). CONCLUSION: The application of ICG for para-aortic SLN mapping should further be investigated and validated in staging surgically locally advanced cervical cancer and those with suspicious lymph nodes on imaging.

Stratified Mucin-producing Intraepithelial Lesions of the Cervix: Clinical Diversity of Cases and Literature Review.

BACKGROUND: This report discusses the current literature on both non-invasive and invasive SMILE ((i)SMILE) lesions of the cervix with focus on the pathology and its related clinical diversity in several cases. Currently, the knowledge on (i)SMILE is limited to single case reports and series. As a consequence, consensus guidelines regarding the management are lacking. Although there is overlap with both high grade squamous intra-epithelial lesion (HSIL) and adenocarcinoma in situ (AIS) on immunohistochemical analyses, it is recommended to treat SMILE like AIS and further excision is needed when surgical margins are positive for SMILE on conization. (i)SMILE, should be considered as a rare subtype of adenocarcinoma of the cervix, and should be treated as such. CASE REPORT: We describe a case with a SMILE lesion undergoing a subsequent robotic hysterectomy after conization and two cases with iSMILE: one case with an early FIGO-stage 1B1 iSMILE tumor, undergoing a robotic radical hysterectomy with sentinel procedure, and one case undergoing a robotic-assisted pelvic/para-aortic lymph node staging dissection, confirming a metastatic FIGO-stage 3C2 (for primary chemo-radiotherapy treatment). CONCLUSION: Here, we report for the first time a few cases of (i)SMILE with different clinical presentations, their management and follow-up. Immunohistochemical characteristics are given for both primary lesions as well as the metastases.

The clinical and financial impact of introducing robotic-assisted hysterectomy in a tertiary referral centre: A direct cost analysis of consecutive hysterectomies during a decade.

BACKGROUND: Economic data and the clinical impact of introducing robotic-assisted hysterectomy in a European setting are scarce with conflicting findings. METHODS: In this retrospective cohort study, the cost and complication rate of the different approaches of hysterectomy are investigated, both benign and (pre)malignant indications were included. RESULTS: 844 patients were included: 323 (38.3%) patients underwent robotic-assisted hysterectomy (RAH), 317 (37.5%) total abdominal hysterectomy (TAH), and 204 (24.2%) total laparoscopic hysterectomy (TLH). TAH dropped from 67.2% to 25.5% of procedures, whilst RAH rose to 41.8% of cases. The total hospitalisation cost was for RAH €5208.39 (±€916.91), for TAH €5846.61 (±€4464.37) and for TLH €3790.06 (±€1267.05). The postoperative complication rate of TAH (9.1%) was significantly higher in comparison with TLH and RAH (5.4% and 3.1%, respectively, p = 0.005). CONCLUSIONS: RAH has replaced TAH in most cases, especially for large uteri, early-stage endometrial cancer and in selected endometriosis cases, resulting in reduced morbidity with lower hospitalisation costs. The indications for TLH remained, including menorrhagia, adenomyosis and persistent cervical dysplasia.

Observational BGOG Study of the Results of Robot-assisted Laparoscopy in 166 Patients with FIGO 2009 Stage IA1-IB1 Cervical Cancer.

STUDY OBJECTIVE: Two recent studies (the Laparoscopic Approach to Cervical Cancer [LACC] trial and a cohort study based on the National Cancer Database) raise the question of whether minimally invasive surgery (conventional and robot-assisted laparoscopy) is inferior to open abdominal surgery in early-stage cervical cancer. In the laparotomy group of the LACC trial, the low rates of recurrence and death are notable. The present study wants to elucidate the current situation of patients with early-stage cervical cancer treated with robot-assisted laparoscopy in hospitals of the Belgium and Luxembourg Gynaecological Oncology Group (BGOG). DESIGN: This is a prospective follow-up study. SETTING: The combined data obtained from different BGOG hospitals were analyzed regarding patients similar to those included in the LACC trial in terms of cervical cancer recurrence and survival. PATIENTS: We included patients with stage IA1, IA2, or IB1 cervical cancer with a histologic subtype of squamous cell carcinoma, adenocarcinoma, or adenosquamous carcinoma. INTERVENTIONS: All patients were treated with robot-assisted laparoscopy. MEASUREMENTS AND MAIN RESULTS: The outcomes were disease-free and overall survival at 3 and 5 years after surgery. A total of 270 patients were included, and 166 were found suitable for analysis. The median age was 45 years. Most patients had International Federation of Gynecology and Obstetrics stage IB1 cervical cancer (84.9%) and squamous cell carcinoma as the histologic subtype (71.7%). The median follow-up time was 44 months, with a range of between 1 and 131 months. Twenty-one recurrences and 12 deaths were noted. Of the deaths, 8 were related to cervical cancer. Disease-free survival was 86% at 3 years (95% confidence interval [CI], 78.52-90.80) and 85% at 5 years (95% CI, 77.03-89.95). Overall survival was 96% at 3 years (95% CI, 90.11-98.22) and 91% at 5 years (95% CI, 82.54 95.17). CONCLUSION: The results of this BGOG study show disease-free and overall survival rates after robot-assisted laparoscopy in early-stage cervical cancer that are at least similar to previous reported recurrence and survival data. We expect that the results of the Robot-assisted Approach to Cervical Cancer trial will elucidate the place of robot-assisted laparoscopy in early-stage cervical cancer.

Robotic Hysterectomy Using a Sealing Device: Differences In Complications and Pain Scores Between Small and Large Uteri.

INTRODUCTION: Robotic hysterectomy using standard diathermy instruments has been reported in benign gynaecology over the past decade, however, perioperative outcomes, such as pain scores, for this procedure between small and large uteri using a sealing device are not well reported in large series. Hence, we aimed to investigate, in a large retrospective cohort study of 156 consecutive patients, the rate of complications between small and large uteri in robotic benign hysterectomy using a sealing device which enables simultaneous dissection, sealing, and cutting of tissues. As secondary outcomes, we investigated the differences in pain scores, operative times, and lengths of hospitalisation between the two groups. To differentiate between small and large uteri, the commonly used threshold of 250 grams was used. Eighty-two cases were included in the small uteri group and 70 in the large uteri group. For four cases, no weight was reported. RESULTS: There was a statistically significant longer operative time for the large uteri group-105 minutes versus 85 minutes for the small uteri group. This difference was due to the longer duration of vaginal removal. The postoperative pain scores were comparable between small and large uteri. The overall complication rate in our series was very low, with 3.2% intraoperative complications and 4.4% postoperative complications. There was no significant difference in peri- and postoperative complication rate, as well as length of hospitalisation between the small and large uteri group. CONCLUSION: In this retrospective cohort study of robotic hysterectomy using a sealing device, there was no difference in perioperative outcome between small and large uteri, but we could demonstrate shorter operative times compared to previous reports on large uteri, most likely due to the versatility of the application of the EndoWrist® Vessel Sealer (Intuitive Surgical, Inc., Sunnyvale, California). Given the known difference in complication rate and length of hospitalization between small and large uteri in laparoscopic hysterectomy, this study shows that robotic hysterectomy is a recommended procedure for benign indications in small, and especially large uteri.

Robot-Assisted Radical Hysterectomy in Cervical Carcinoma: The Belgian Experience.

OBJECTIVE: The purpose of this study was to report the experience and oncological outcome of robot-assisted radical hysterectomies (RRHs) for cervical cancer performed in Belgium. METHODS: Patients undergoing RRH for cervical cancer (n = 109) were prospectively collected between July 2007 and April 2014 in the 5 Belgian centers performing RRH for cervical cancer. RESULTS: The median age of the patients was 46 years (range, 31-80 years). Histological types included squamous cell carcinoma in 61 patients, adenocarcinoma in 22 patients, adenosquamous in 8 patients, endometrioid carcinoma in 2 patients, and other types (n = 16). The International Federation of Gynecology and Obstetrics stage distribution was IA (n = 9), stage IB1 (n = 71), stage IB2 (n = 4), stage II (n = 24), and unknown (n = 1). Twenty-four patients received adjuvant therapy, 17 patients underwent radiochemotherapy, and 7 underwent adjuvant radiation. Eighteen patients relapsed, and 5 died of disease. The median follow-up was 27.5 months (range, 3-82 months). The 2- and 5-year overall survivals were 96% and 89%, respectively. The 2- and 5-year disease-free survivals (DFSs) were 88% and 72%, respectively. The 2-year DFS per stage was 100% for IA, 88% for IB1, 100% for IB2, and 83% for II. The 5-year DFS per stage was 100% for stage IA and 75% for IB1. The complications were as expected for radical hysterectomy. CONCLUSIONS: This series confirms the feasibility and safety of RRH not only in cervical cancer stage IA to IB1, but also after neoadjuvant chemotherapy in stage IB2 to IIB.

Robotic hysterectomy using the Vessel Sealer for myomatous uteri: technique and clinical outcome.

OBJECTIVE: Robotic procedures using the Vessel Sealer are not well reported in the literature, especially given the advantages of sealing devices already studied in standard laparoscopic procedures. This study reports our experience with the EndoWrist(®) One™ Vessel Sealer in robotic hysterectomy for myomatous uteri. STUDY DESIGN: In this retrospective cohort study of the first 50 consecutive patients with myomatous uteri undergoing a robotic hysterectomy, we report our experience with the EndoWrist(®) One™ Vessel Sealer (Intuitive Surgical Inc., Sunnyvale, CA) during this procedure. The learning curve was evaluated, and the operative times as well as the complications were recorded. RESULTS: After the first 10 cases, the median console and total (skin-to-skin) operative time dropped significantly from 110 to 60min and from 158 to 105min, respectively (p=0.018 and p=0.008 respectively). The body mass index (≤ or >30kg/m(2)), uterine weight (≤ or >250g), and uterine size had no statistical significant effect on the total operative time. Median blood loss during surgery was 63mL in all cases (range: 0-400mL). The morbidity was low, and approximately 50% of cases could be discharged from the hospital after one to two days. CONCLUSION: Robotic hysterectomy using the Vessel Sealer has, after a short learning curve of 10 cases, similar operative times than other published reports on robotic hysterectomy or laparoscopic hysterectomy using a sealing device for myomatous or large uteri.

An unusual cervical tumor as presentation of a non-hodgkin lymphoma.

Rare cervical cancers are responsible for a minority of cases encountered by a clinician. However, behavioral patterns, management, and prognosis of certain rare cervical cancers differ from either squamous carcinomas or adenocarcinomas. Here we present a case of a locally advanced cervical tumor as a presentation of an extranodal cervical non-Hodgkin lymphoma (NHL), with a review of the current literature.

The absence of high-risk HPV combined with specific p53 and p16INK4a expression patterns points to the HPV-independent pathway as the causative agent for vulvar squamous cell carcinoma and its precursor simplex VIN in a young patient.

Simplex vulvar intraepithelial neoplasia (VIN) is an important precursor of vulvar invasive squamous cell carcinoma and characteristically occurs in postmenopausal women. In this report, the absence of high-risk human papillomavirus (HPV) combined with specific p53 and p16INK4a expression patterns points to the HPV-independent pathway as the causative agent for vulvar squamous cell carcinoma in a 28-year-old woman. Its precursor simplex VIN was initially interpreted as eczema. Although simplex VIN has a predilection for postmenopausal women, it can occur in young patients. The development of invasive vulvar squamous cell carcinoma underlines the importance of including simplex VIN in the differential diagnosis of vulvar lesions, even at a young age. Furthermore, knowledge about the HPV status in the tumor and thus the underlying causative pathway can alert the gynecologist for the presence or absence of multicentric lower genital tract disease, as this is frequent in the HPV-dependent and not in the HPV-independent pathway.

Lymphangiogenesis in human gynaecological cancers.

The metastatic spread of tumor cells is responsible for the majority of cancer deaths, and with few exceptions, all cancers can metastasize. Clinical findings have for a long time suggested that by providing a pathway for tumor cell dissemination, tumor-associated lymphatics act as key components of metastatic spread. This is believed to occur principally via pre-existing and possibly also newly formed lymphatics (lymphangiogenesis). Increased expression of vascular endothelial growth factor-C (VEGF-C) and VEGF-D in primary tumors correlates with increased dissemination of tumor cells to regional lymph nodes (LNs) in a variety of human carcinomas. Here we will review the mechanisms of lymphangiogenesis, particularly in the context of metastatic tumor spread, and will critically examine the role of VEGF-C and VEGF-D in this process in gynaecological cancers. Potential anti-lymphangiogenic strategies are also discussed.

Expression of vascular endothelial growth factor (VEGF)-C and VEGF-D, and their receptor VEGFR-3, during different stages of cervical carcinogenesis.

Cervical carcinogenesis has well-defined stages of disease progression including three grades of pre-invasive lesions--cervical intraepithelial neoplasia grades 1-3 (CIN 1-3)--and invasive cervical cancer. However, the biological properties of CIN lesions prone to develop invasive disease are not well defined. Recent observations suggest that early invasive disease spreads to regional lymph nodes in several tumour types and that growth factors (VEGF-C and VEGF-D) involved in new lymphatic vessel formation may play a crucial role in this process. The present study has assessed the expression of VEGF-C and VEGF-D, and their receptor VEGFR-3, in 152 cervical lesions (33 CIN 1, 33 CIN 2, 37 CIN 3, and 49 squamous cell carcinomas) to determine whether expression of lymphangiogenic factors occurs prior to invasion. The presence of lymphatic vessels was determined using LYVE-1 and podoplanin staining, as well as double immunostaining for LYVE-1/CD34 and podoplanin/CD34. In situ hybridization was performed to determine VEGFR-3 mRNA expression. A significant positive correlation was found between VEGF-C, VEGF-D, and VEGFR-3 expression through the different stages of cervical carcinogenesis. Significant differences in protein expression for VEGF-C, VEGF-D, and VEGFR-3 were found between CIN 1-2 and CIN 3 (p<0.001 for all), but not between CIN 3 and cervical cancer. More than 50% of the CIN 3 lesions showed moderate to strong staining for VEGF-C and VEGF-D, whereas most of the early pre-cancerous lesions (CIN 1 and 2) were negative. In cervical cancer, similar observations to those in CIN 3 were found. VEGFR-3 mRNA expression was found in the cytoplasm of epithelial neoplastic cells and VEGFR3 protein expression was found in more than 50% of CIN 3 lesions and cervical cancers, compared with 15% in CIN 1 and 2. These findings suggest an autocrine growth stimulation pattern via VEGFR-3. Adjacent CIN 3 was present in nine cervical cancers and displayed strong expression for VEGF-C, VEGF-D, and VEGFR-3. These results suggest that in cervical carcinogenesis a switch to the lymphangiogenic phenotype may occur at the stage of CIN 3.

Combination of microdissection and microarray analysis to identify gene expression changes between differentially located tumour cells in breast cancer.

Comparison of gene expression changes between cancer cells at the periphery and in the centre of breast cancers was performed using a combination of microdissection and microarray analysis. Cancer cells from the two areas were pooled separately from five patients with ductal carcinoma in situ and separately from five patients with frankly invasive cancer. Limited total RNA, 100-200 ng, from this microdissected tissue required use of the Atlas SMART trade mark Probe Amplification Kit to synthesize and amplify cDNA and make (33)P-labelled probes. Probes were then hybridized to Atlas Human Cancer 1.2 Arrays containing 1176 known genes. Triplicate analysis revealed that 22 genes changed their expression levels in the periphery relative to the central region: 15 upregulated and seven downregulated (arbitrary threshold of 1.5-fold or greater). Differences in RNA levels were confirmed by quantitative real-time PCR for two of the genes and by changes in protein levels, detected by immunohistochemistry, for a couple of representative gene products. Thus, changes in gene expression associated with variation in microanatomical location of neoplastic cells can be detected within even small developing tumour masses.

Characterization of adnexal mass lesions on MR imaging.

OBJECTIVE: The aim of our study was to evaluate the accuracy of MR imaging in the detection and characterization of adnexal mass lesions and to determine which imaging features are predictive of malignancy. SUBJECTS AND METHODS: We prospectively performed MR imaging in 104 patients (age range, 19-87 years; mean age, 50 years) with clinically or sonographically detected complex adnexal masses. We used a 1.5-T unit to perform T1-, T2-, and fat-suppressed T1-weighted sequences before and after IV injection of gadolinium. The adnexal lesions were examined for several features including size, shape, character (solid-cystic), vegetation, signal intensity, and enhancement. Secondary signs such as ascites, peritoneal disease, and lymphadenopathy were noted. We compared the imaging features with the surgical and pathologic findings. Multiple logistic regression analysis was performed on all MR imaging features. RESULTS: A total of 163 lesions--94 benign and 69 malignant lesions--were examined. On MR imaging, 95% (155/163) of the lesions were detected. The overall accuracy for the diagnosis of malignancy was 91%. On univariate analysis, the imaging features associated with malignancy were a solid-cystic lesion, irregularity, and vegetation on the wall and septum in a cystic lesion, the large size of the lesion, an early enhancement on dynamic contrast-enhanced MR images, and the presence of ascites, peritoneal disease, or adenopathy. On multiple logistic regression analysis, ascites and vegetation in a cystic lesion were the factors most significantly indicative of malignancy. CONCLUSION: MR imaging is highly accurate in the characterization of adnexal mass lesions, and the best predictors of malignancy are vegetation in a cystic lesion and ascites.