Prognostic value of putative circulating cancer stem cells in patients undergoing hepatic resection for colorectal liver metastasis.

BACKGROUND: Although surgery is the gold standard treatment of hepatic metastasis from colorectal cancer (CRC), many patients ultimately die of their disease. We tested the hypothesis that the detection of circulating tumor cells (CTC) might identify patients at high risk of dying of disease recurrence after apparently radical liver surgery. METHODS: We considered 50 patients undergoing radical surgery for liver-confined hepatic metastasis from CRC. The expression of a panel of cancer-related genes, as assessed by quantitative real-time PCR, was used to detect CTC in the peripheral blood of these patients immediately before surgery. Survival analysis was performed by the Cox regression model. RESULTS: Univariate analysis of the expression levels of CD133 (a marker of colon cancer stem cells) and survivin (an antiapoptotic factor) resulted in statistically significant association with patient survival [hazard ratio (HR) 2.7, 95% confidence interval (CI) 1.9-3.7, P < 0.0001; and hazard ratio 2.1, 95% CI 1.4-3.2, P < 0.0001, respectively]. Remarkably, multivariate analysis found that only the transcriptional amount of CD133 resulted in statistical significance (HR 2.6, 95% CI 1.9-3.6, P < 0.0001), indicating that this biomarker can independently predict the survival of these patients. CONCLUSIONS: CD133-positive CTC may represent a suitable prognostic marker to stratify the risk of patients who undergo liver resection for CRC metastasis, which opens the avenue to identifying and potentially monitoring the patients who are most likely to benefit from adjuvant treatments.

The prognostic value of serum S100B in patients with cutaneous melanoma: a meta-analysis.

S100B protein detected in the serum of patients with cutaneous melanoma has been long reported as a prognostic biomarker. However, no consensus exists on its implementation in the routine clinical setting. This study aimed to comprehensively and quantitatively summarize the evidence on the suitability of serum S100B to predict patients' survival. Twenty-two series enrolling 3393 patients with TNM stage I to IV cutaneous melanoma were reviewed. Standard meta-analysis methods were applied to evaluate the overall relationship between S100B serum levels and patients' survival (meta-risk). Serum S100B positivity was associated with significantly poorer survival (hazard ratio [HR] = 2.23, 95% CI: 1.92-2.58, p < 0.0001). Between-study heterogeneity was significant, which appeared to be related mainly to dissemination bias and the inclusion of patients with stage IV disease. Considering stage I to III melanoma (n = 1594), the meta-risk remained highly significant (HR = 2.28, 95% CI: 1.8-2.89; p < 0.0001) and studies' estimates were homogeneous. Subgroup analysis of series reporting multivariate survival analysis supported S100B as a prognostic factor independent of the TNM staging system. Our findings suggest that serum S100B detection has a clinically valuable independent prognostic value in patients with melanoma, with particular regard to stage I-III disease. Further investigation focusing on this subset of patients is justified and warranted before S100B can be implemented in the routine clinical management of melanoma.

Correlation between magnetic resonance imaging and histopathological tumor response after neoadjuvant chemotherapy in breast cancer.

AIM: To evaluate the accuracy of magnetic resonance imaging in assessing tumor response following neoadjuvant chemotherapy in patients with locally advanced breast cancer. MATERIALS AND METHODS: Twenty-six patients entered a phase II study of neoadjuvant chemotherapy, undergoing bilateral breast magnetic resonance imaging before therapy and before surgery. Tumor response was classified using RECIST criteria, using tumor size at magnetic resonance imaging. The latter was then compared to residue found at histopathological examination. RESULTS: Magnetic resonance imaging showed 6 (23%) complete responses, 17 (65%) partial responses, 3 (11.5%) disease stabilizations and no disease progressions. Twenty-three tumors (88.5%) were considered responsive and 3 (11.5%) unresponsive. Pathological tumor response was: 6 complete responses (23%), 17 partial responses (65%), 2 stable disease (8%), 1 progression (4%). When results of the preoperative magnetic resonance imaging were compared to pathological tumor response, magnetic resonance imaging overestimated tumor size in 12 cases (46%) and underestimated it in 9 (35%). However, preoperative magnetic resonance imaging failed to detect invasive tumor in 2 false-negative cases (8%), 1 of which was multifocal. Mastectomy was performed in 12 cases: 1 case of disease progression even though the neoplasm appeared smaller at magnetic resonance imaging, 3 cases with stable disease, and 4 cases with T3 or T4 disease. The 9th patient was T2N2 with initial retroareolar disease and negative magnetic resonance imaging after chemotherapy. The 10th patient, affected by lobular cancer, was in partial remission but was T3N1. The 11th patient was 57 years old but was not interested in conservative surgery. The 12th patient requested bilateral prophylactic mastectomy due to her positive family history of breast cancer. CONCLUSIONS: Magnetic resonance imaging of the breast allowed conservative surgery in 54% of the patients. This low value is primarily due to overestimation of tumor size, with a negative predictive value of 67% in our population. However, surgeons were able to choose conservative surgery with relative safety in cases of small residual disease.

Sentinel lymph node metastasis from mammary ductal carcinoma in situ with microinvasion.

Axillary lymph node dissection (ALND) in patients with ductal carcinoma in situ with microinvasion (DCISM) of the breast was controversial, because of the relevant morbidity incurred by the procedure and the low incidence of axillary involvement. The introduction of the sentinel lymph node (SLN) biopsy as a minimally invasive procedure for axillary staging has prompted new interest in this issue. However, as DCISM is a rare type of cancer, data on the incidence of SLN metastasis are scarce. The aim of the present paper was therefore to assess the prevalence of SLN metastasis in a multi-institutional series of DCISM patients, in order to ascertain whether SLN biopsy might be justified as a standard procedure in the presence of microinvasive cancer. Between 1999 and 2004, 43 patients with a diagnosis of DCISM underwent SLN biopsy. Microinvasion was defined as one or more foci of invasion beyond the basal membrane, none exceeding 1mm. SLNs were examined following haematoxylin-eosin and immunohistochemical staining. SLN metastases were found in four out of 43 cases (9.3%). In one patient, SLN contained only micrometastasis. All four patients with positive SLN underwent complete ALND and in all these cases further metastatic axillary nodes were found. In conclusion, given the relevant incidence of nodal metastases and the low morbidity of the procedure, we believe that SLN biopsy should be considered in all patients with a diagnosis of DCISM. In cases of SLN involvement, even if micrometastatic, our policy is to perform a complete ALND.

Role of axillary sentinel lymph node biopsy in patients with pure ductal carcinoma in situ of the breast.

BACKGROUND: Sentinel lymph node (SLN) biopsy is an effective tool for axillary staging in patients with invasive breast cancer. This procedure has been recently proposed as part of the treatment for patients with ductal carcinoma in situ (DCIS), because cases of undetected invasive foci and nodal metastases occasionally occur. However, the indications for SLN biopsy in DCIS patients are controversial. The aim of the present study was therefore to assess the incidence of SLN metastases in a series of patients with a diagnosis of pure DCIS. METHODS: A retrospective evaluation was made of a series of 102 patients who underwent SLN biopsy, and had a final histologic diagnosis of pure DCIS. Patients with microinvasion were excluded from the analysis. The patients were operated on in five Institutions between 1999 and 2004. Subdermal or subareolar injection of 30-50 MBq of 99 m-Tc colloidal albumin was used for SLN identification. All sentinel nodes were evaluated with serial sectioning, haematoxylin and eosin staining, and immunohistochemical analysis for cytocheratin. RESULTS: Only one patient (0.98%) was SLN positive. The primary tumour was a small micropapillary intermediate-grade DCIS and the SLN harboured a micrometastasis. At pathologic revision of the specimen, no detectable focus of microinvasion was found. CONCLUSION: Our findings indicate that SLN metastases in pure DCIS are a very rare occurrence. SLN biopsy should not therefore be routinely performed in patients who undergo resection for DCIS. SLN mapping can be performed, as a second operation, in cases in which an invasive component is identified in the specimen. Only DCIS patients who require a mastectomy should have SLN biopsy performed at the time of breast operation, since in these cases subsequent node mapping is not feasible.

Sentinel node biopsy for breast cancer: is it already a standard of care? A survey of current practice in an Italian region.

BACKGROUND: Although sentinel node biopsy (SNB) is becoming the standard approach for axillary staging in patients with small breast cancer, criteria for patient selection and some technical aspects of the procedure have yet to be clearly defined. The aim of the present survey was therefore to investigate the way in which SNB is used by general surgeons working in the Veneto region, Italy. METHODS: A 29-item questionnaire regarding various aspects of SNB practice was mailed to surgeons in charge of breast surgery in all the 56 surgical centres of the region. RESULTS: The rate of response to the questionnaire was 82.1% (n = 46); 69.6% (n = 32) of the respondents routinely perform SNB in their clinical practice. Most of the interviewed surgeons (93.5%) expressed the belief that the acceptable false negative rate should be < or =5%. However, among the surgeons who perform SNB, only 34.4% performed more than 20 SNB during the learning phase. Indications are limited to tumours of < or =1 cm by 31.2% (n = 10) of respondents, < or =2 cm by 46.9% (n = 15) and < or =3 cm by 21.9% (n = 7). Almost all respondents (93.7%) agreed that a clinically positive axilla is a contraindication to SNB, while opinions differed widely concerning other potential contraindications. In most of the centres considered, SN identification is undertaken on the day before surgery using a subdermal injection of 30-50 MBq of 99mTc-albumin-nanocolloid followed by lymphoscintigraphy. CONCLUSIONS: SNB is currently performed in the majority of hospitals in the Veneto region. However, the training phase and criteria used for patient selection differ from centre to centre. Certified training courses and shared guidelines are therefore highly desirable.

Subareolar injection for sentinel lymph node location in breast cancer.

AIM: Several different injection techniques are currently used for sentinel node (SN) identification in patients with breast cancer. Some studies suggest that the subareolar plexus drains lymph from the whole breast to the same axillary SN. In order to test this hypothesis, we ascertained whether subareolar blue dye injection and subdermal radioisotope injection close to the tumour identify the same axillary nodes. METHODS: One day prior to surgery, 50 patients with breast cancer underwent subdermal injection of 30-40MBq of 99m-Tc colloidal albumin (Nanocoll) at the site of the cutaneous projection of the tumour. Ten minutes before surgery, each patient received a subareolar injection of 2-3cc of patent blue. All axillary radioactive nodes and blue-stained nodes were excised and a histologic examination was made. RESULTS: Radioisotope marked the SNs in 47/50 (94%) cases, and the blue dye in 43/50 cases (86%). In three cases, SNs were not identified with either method. Of the 43 cases in which both the tracers reached the axilla, in 40 (93%) the SN was hot and blue-stained, while in 3 cases the two tracers identified different nodes. CONCLUSIONS: Our findings suggest that subareolar injection and subdermal injection elsewhere in the breast usually identify the same SN. Subareolar injection appears to be particularly valuable in patients with multicentric or deep non-palpable breast tumours.

Analysis of technical and clinical variables affecting sentinel node localization in patients with breast cancer after a single intradermal injection of 99mTc nanocolloidal albumin.

AIM: To investigate the technical, clinical and pathological findings that can, potentially, affect pre-operative lymphoscintigraphy in visualizing sentinel lymph node (SLN) and intra-operative probe detection of SLN in patients with breast cancer. METHODS: One hundred and forty-two consecutive female patients with, clinically, a solitary, small breast cancer and clinically N0 axilla were enrolled. Preoperative lymphoscintigraphy was performed by a single intradermal injection of 99mTc nacolloidal albumin (Nanocoll) the day before surgery. For radioguided surgery two gamma probes with diameters of 11 mm and 15 mm, and set up with a count rate ranging from 1 to 4 s were used. The following variables were evaluated: patient's age, radiotracer dose, volume of injectate, primary tumour location, primary tumour size, and presence and extension of axillary nodal metastases. RESULTS: Lymphoscintigraphy showed high sensitivity in visualizing the SLN (98% success rate) and it resulted in a rapid technique since SLN was visualized within 30 min from injection in 85.21% of cases for the whole series. The probe detection rate was also very high (97.8% success rate): the mean per cent uptake in the SLN was 0.98. Statistical analysis showed that no parameter was found to have significantly influenced either SLN visualization at lymphoscintigraphy or SLN probe detection at surgery. CONCLUSION: In our experience, lymphoscintigraphy performed by a single intradermal injection of Nanocoll was an effective and rapid technique for visualizing axilla SLNs in breast cancer patients. Moreover, this technique appeared to be independent of any technical, clinical and pathological findings.

Phase II study of neoadjuvant gemcitabine, pegylated liposomal doxorubicin, and docetaxel in locally advanced breast cancer.

AIM: This was a phase II study to assess the activity of a novel neoadjuvant regimen in locally-advanced breast cancer. PATIENTS AND METHODS: Fifty patients with histological confirmation of locally advanced breast cancer received treatment with gemcitabine 1000 mg/m(2) (day 1) followed by gemcitabine 800 mg/m(2) plus docetaxel 75 mg/m(2) plus pegylated liposomal doxorubicin (PLD) 30 mg/m(2) (day 8) every 3 weeks for at least 4 cycles, plus a final 2 additional cycles. Tumour size was T1 (n=2), T2 (n=32), T3 (n=14), T4 (n=2). All 50 patients underwent surgery. RESULTS: Clinical complete, partial and no response were observed in 13 (26%), 24 (48%) and 11 (22%) patients, respectively (overall response rate: 74%). The number of chemotherapy cycles was found to be an independent predictor of a pathologic complete response. CONCLUSION: The combination of gemcitabine-docetaxel-PLD can yield high tumour response rates in patients with locally-advanced breast cancer who undergo a full treatment of 6 cycles.

Role of resection margins in patients treated with breast conservation surgery.

BACKGROUND: After breast conservation therapy (BCT), margin status is routinely evaluated to select patients who need reexcision. The aim of this study was to investigate how margin status and other clinicopathologic factors correlate with the presence of residual tumor at reexcision. METHODS: A series of 431 breast cancer patients who underwent BCT followed by reexcision were considered because they had positive or close (< or =3 mm) margins. At univariate and multivariate analysis the frequency of residual tumor in the reexcision specimens was associated with the status and width of resection margins and with a series of other clinicopathologic factors. RESULTS: Of the 382 evaluable patients, 253 had positive and 129 close margins. Residual tumor was found at reexcision in 51.8% positive-margin patients and 34.1% close-margin patients (P = .001). In the latter group tumor-margin distance (range, 0.08 to 3 mm) was not associated with the incidence of residual tumor (P = .134). On univariate analysis age < or =45 years (P = .006), intraductal histotype (P = .005), size > 2 cm (P = .010), positive axillary nodes (P = .031), and timing of reexcision (P = .044) were significantly associated with a higher frequency of residual tumor. All these factors, except tumor size, maintained a significant predictive value on multivariate analysis. CONCLUSIONS: In the presence of positive margins, relevant residual disease cannot be ruled out and further surgery is indicated. Close margins do not mandate reexcision because they may indicate either that the tumor has been radically excised or the presence of residual foci of a multifocal tumor, which are usually effectively treated by radiotherapy.

A Randomized clinical trial on sentinel lymph node biopsy versus axillary lymph node dissection in breast cancer: results of the Sentinella/GIVOM trial.

OBJECTIVE: The aim of this multicenter randomized trial was to assess the efficacy and safety of sentinel lymph node (SLN) biopsy compared with axillary lymph node dissection (ALND). BACKGROUND: All studies on SLN biopsy in breast cancer report a variable false negative rate, whose prognostic consequences are still unclear. METHODS: From May 1999 to December 2004, patients with breast cancer < or =3 cm were randomly assigned to receive SLN biopsy associated with ALND (ALND arm) or SLN biopsy followed by ALND only if the SLN was metastatic (SLN arm). The main aim was the comparison of disease-free survival in the 2 arms. RESULTS: A total of 749 patients were randomized and 697 were available for analysis. SLNs were identified in 662 of 697 patients (95%) and positive SLNs were found in 189 of 662 patients (28.5%). In the ALND group, positive non-SLNs were found in 18 patients with negative SLN, giving a false negative rate of 16.7% (18 of 108). Postoperative side effects were significantly less in the SLN group and there was no negative impact of the SLN procedure on psychologic well being. At a median follow-up of 56 months, there were more locoregional recurrences in the SLN arm, and the 5-year disease-free survival was 89.9% in the ALND arm and 87.6% in the SLN arm, with a difference of 2.3% (95% confidence interval: -3.1% to 7.6%). However, the number of enrolled patients was not sufficient to draw definitive conclusions. CONCLUSION: SLN biopsy is an effective and well-tolerated procedure. However, its safety should be confirmed by the results of larger randomized trials and meta-analyses.

Support vector machine learning model for the prediction of sentinel node status in patients with cutaneous melanoma.

BACKGROUND: Currently, approximately 80% of melanoma patients undergoing sentinel node biopsy (SNB) have negative sentinel lymph nodes (SLNs), and no prediction system is reliable enough to be implemented in the clinical setting to reduce the number of SNB procedures. In this study, the predictive power of support vector machine (SVM)-based statistical analysis was tested. METHODS: The clinical records of 246 patients who underwent SNB at our institution were used for this analysis. The following clinicopathologic variables were considered: the patient's age and sex and the tumor's histological subtype, Breslow thickness, Clark level, ulceration, mitotic index, lymphocyte infiltration, regression, angiolymphatic invasion, microsatellitosis, and growth phase. The results of SVM-based prediction of SLN status were compared with those achieved with logistic regression. RESULTS: The SLN positivity rate was 22% (52 of 234). When the accuracy was > or = 80%, the negative predictive value, positive predictive value, specificity, and sensitivity were 98%, 54%, 94%, and 77% and 82%, 41%, 69%, and 93% by using SVM and logistic regression, respectively. Moreover, SVM and logistic regression were associated with a diagnostic error and an SNB percentage reduction of (1) 1% and 60% and (2) 15% and 73%, respectively. CONCLUSIONS: The results from this pilot study suggest that SVM-based prediction of SLN status might be evaluated as a prognostic method to avoid the SNB procedure in 60% of patients currently eligible, with a very low error rate. If validated in larger series, this strategy would lead to obvious advantages in terms of both patient quality of life and costs for the health care system.

Number of metastatic sentinel nodes as predictor of axillary involvement in patients with breast cancer.

BACKGROUND AND OBJECTIVES: More than half of patients with positive sentinel node (SN) have no metastases in non-sentinel nodes (NSNs) on axillary lymph node dissection (ALND). The aim of this study was to investigate factors predictive of NSNs involvement, in order to identify patients with metastatic disease confined to the SN which might avoid ALND. METHODS: ALND was performed in 167 patients with metastatic SN. Axillary NSNs status was correlated with the size of SN metastases, the size of the primary tumor and the occurrence of lymphovascular invasion. In 72 cases, the radiotracer (Tc-99m albumin colloid) marked multiple (in most cases 2 or 3) nodes. In this group, NSNs status was correlated with the number of metastatic radioactive nodes (1 or > 1), and with the above mentioned histopathologic factors. RESULTS: NSNs metastases were found in 57/167 cases (34.1%), the rate increasing proportionate to the size of both SN metastases (p < 0.0001) and primary tumor (p = 0.0075), while no significant correlation was found for lymphovascular invasion (p = 0.1769). At univariate and multivariate analysis of findings from the 72 cases with multiple probe-detected hot nodes, positivity in more than one hot node was the strongest predictor of NSN involvement (p = 0.0019). CONCLUSIONS: The identification and excision of multiple hot nodes can be useful in the prediction of NSNs involvement in patients with metastatic SN.

The role of preoperative ultrasound scan in detecting lymph node metastasis before sentinel node biopsy in melanoma patients.

BACKGROUND AND OBJECTIVES: To evaluate the efficacy of preoperative ultrasound (US) scanning in identifying lymph node metastasis before sentinel node biopsy (SNB), we conducted a prospective study on 125 patients with primary cutaneous melanoma (CM). METHODS: We prospectively enrolled 125 patients with >1 mm thick CM and candidate for SNB. Preoperatively, patients underwent US scanning of regional lymphatic basins and FNA of suspected lymph nodes (LN). All patients underwent lymphatic mapping and SNB. RESULTS: Combined with fine-needle aspirate (FNA) of suspect LN, US scan allowed the correct preoperative detection of 12 out of 31 histologically positive lymphatic basins, specificity and sensitivity being 100 and 39%, respectively. The false negative rate (61%) was mainly linked to tumor deposits less than 2 mm in diameter, which can be considered the current spatial resolution limit of this technique. CONCLUSIONS: Preoperative US scan could reduce the number of SNB, thus avoiding the stress of this surgical procedure in approximately 10% of patients and reducing health care costs. As a non-invasive and relatively inexpensive technique, lymph node US scan can be part of the preoperative staging process of patients' candidate for SNB in order to avoid unnecessary surgical procedures.