Clinical significance of circulating tumor cells in the response to trastuzumab for HER2-negative metastatic gastric cancer.

PURPOSE: The prognosis of metastatic gastric cancer has improved due to trastuzumab in patients with HER2 positive. Circulating tumor cells (CTCs) have been examined as a prognostic predictor in gastric cancer. The clinical advantage of trastuzumab was examined in gastric cancer patients with HER2-negative tumor tissues and HER2-positive CTCs. METHODS: A total of 105 patients with metastatic or recurrence gastric cancer were enrolled. All patients were examined HER2 expression in CTC using the CellSearch system in blood specimens. RESULTS: CTCs were detected in 65 of 105 patients (61.9%) and 61 patients were divided into three groups: Group A (n = 27), histological HER2-positive; Group B (n = 17), histological HER2-negative and HER2-positive CTCs; and Group C (n = 17), HER2-negative on histology and CTCs. Patients received capecitabine plus cisplatin. Groups A and B were additionally treated by trastuzumab. There was no relationship between tumor tissues and CTCs in HER2 expression. Even if group B had no histological HER2 expression, group B showed a good prognosis as same as group A, and group C had a significantly worse overall survival than groups A and B. The multivariate analysis demonstrated that HER2-expression on CTCs was an independent prognostic factor for both overall and progression-free survival. CONCLUSION: The present results indicate the potential clinical utility of trastuzumab combined chemotherapy in patients with HER2-positive CTCs even if they are histologically HER2-negative.

Biological aggressiveness of alpha-fetoprotein (AFP)-positive gastric cancer.

BACKGROUND/AIMS: Alpha-fetoprotein (AFP)-positive gastric cancer (APGC) which occupied well-defined gastric cancer entity, reportedly has an aggressive behavior with hematogenous metastasis. However, little information regarding the clinicopathological and biological behaviors of APGC is available due to the small size of reported series. METHODOLOGY: We retrospectively analyzed the clinical features of APGC in 556 patients with gastric cancer who underwent preoperative measurement of serum AFP levels and gastrectomy in Kagoshima University Hospital. APGC was regarded as any cancer with preoperative serum AFP levels above the cutoff level of 5 ng/mL. Clinicopathological features of APGC were assessed using the General Rules of Gastric Cancer. Of the 556 patients, 97 patients underwent immunohistochemical evaluation of AFP expression in the primary tumor. Both p53 and MIB-1 expression were examined at the same time and compared with AFP expression. Biological aggressiveness of APGC was estimated. RESULTS: Serum AFP positivity was detected in 4.3% of cases (range, 0-2202 ng/mL). Patients were divided into 25 APGC patients and 531 non-APGC patients. APGC displayed deeper tumor invasion, increased nodal involvement, increased venous invasion, and increased CEA concentrations compared to gastric cancer in non-APGC. Surgical outcomes for APGC were significantly worse than those for non-APGC (p < 0.05). All recurrences in patients with APGC involved hepatic metastasis. Abnormalities of p53 were more frequent for APGC than for non-APGC (p < 0.05). CONCLUSIONS: APGC was strongly associated with hematogenous factors such as venous invasion, hepatic metastasis and aggressive biological factors (p53 abnormalities). Considering the aggressive biological behavior of APGC, we must closely follow up for patients with such tumor, including postoperative adjuvant therapy.

Initial metastatic, including micrometastatic, sites of lymph nodes in esophageal squamous cell carcinoma.

BACKGROUND AND OBJECTIVES: It is important to identify the initial lymph node metastasis when performing less invasive surgery. The purpose of the present study was to analyze locations of solitary lymph node metastasis and micrometastasis in esophageal carcinoma. METHODS: We retrospectively analyzed the initial sites of lymph node metastasis in esophageal cancer. Sixty-five consecutive patients with solitary lymph node metastasis, and 33 pN0 patients with only lymph node micrometastasis detected by immunohistochemistry, were classified according to tumor location and tumor depth. RESULTS: The location of lymph node metastasis in the 22 patients with superficial cancer was limited to recurrent nerve nodes (RN) in the upper thoracic esophagus; RN, paraesophageal nodes (PE), or perigastric nodes (PG) in the middle or lower thoracic esophagus. Thirty-six patients with advanced cancer had lymph node metastasis at RN, PE, or PG locations, while in the remaining seven, lymph node metastasis was found in areas far from the primary tumor. Regarding the 33 patients with lymph node micrometastasis, the locations of micrometastasis were similar to those of solitary metastasis. CONCLUSIONS: Although less invasive surgery, such as reduction of lymphadenectomy, may be suitable for superficial cancer, it should be performed with special care in advanced cancer.

CD3 zeta expression of regional lymph node and peripheral blood lymphocytes in gastric cancer.

BACKGROUND AND AIM: Impaired expression of the CD3 zeta chain in T cells associated with T cell anergy has been reported in cancer patients. However, few studies have investigated CD3 zeta expression in regional lymph node lymphocytes (LAL) in cancer patients. This study aims to confirm CD3 zeta expression levels by lymph node and peripheral blood lymphocytes (PBL) in gastric cancer patients and to discuss the clinical implications of intranodal or peripheral blood expression of CD3 zeta in gastric cancer. PATIENTS AND METHODS: Twenty-two gastric cancer patients were enrolled. Macroscopically non-metastatic compartment 1 LNL (C1LNL), compartment 2 LNL (C2LNL) and PBL were surgically obtained. Two-color flow cytometry was then used to quantify the levels of CD3 zeta expression in C1LNL, C2LNL and PBL. RESULTS: Impaired CD3 zeta expression was confirmed in 9.5% of C1LNL, 8.9% of C2LNL and 4.2% of PBL. There was a significant difference in CD3 zeta expression levels between C1LNL and PBL (p<0.01). CD3 zeta expression in PBL was significantly correlated with depth of invasion but not nodal involvement. Distinct differences between the respective lymph node compartments were not identified. CONCLUSION: Immunological paralysis following CD3 zeta impairment may occur more frequently in LNL than in PBL in gastric cancer. Identifying such patients during the perioperative period using flow cytometric methods will increase the efficacy of cytokine therapy aiming to normalize CD3 zeta expression levels.

Bone marrow micrometastasis detected by RT-PCR in esophageal squamous cell carcinoma.

The clinical implications of bone marrow micrometastases (BMM) detected by RT-PCR in esophageal squamous cell carcinoma (ESCC) have not been elucidated. We evaluated the relation between the presence of BMM, both before and after surgery, and clinicopathologic findings in patients with ESCC. Bone marrow samples from 48 patients with ESCC were obtained from the iliac crest before and after surgery. After total RNA was extracted from each bone marrow sample, carcinoembryonic antigen (CEA)-specific RT-PCR was performed. BMM was detected by RT-PCR in 10 of the 48 patients. Four patients each had positive signals only before or only after surgery and 2 patients had positive signals both before and after surgery. There were no significant differences in clinicopathologic factors, including neoadjuvant therapy, between patients with BMM and without BMM. To date, the rates of recurrent disease in patients with BMM and without BMM are 80% (8/10) and 50% (19/38), respectively, a difference which is not significant. The 4-year survival rates of patients with BMM and without BMM are 10.0% and 47.3%, respectively. Recurrence and survival rates were poorer in patients with RT-PCR positivity, although the differences were not significant. A larger study is required to clarify the clinical impact of BMM.

Clinical implications of intratumoral dendritic cell infiltration in esophageal squamous cell carcinoma.

The clinical impact of intratumoral dendritic cell infiltration (IDCI) in esophageal cancer is not fully understood. We investigated the relationship between IDCI and clinical features in esophageal carcinoma. A retrospective analysis of a total of 203 patients who underwent esophagectomy for squamous cell esophageal carcinoma was performed. We evaluated IDCI immunohistochemically using S-100-protein. Under a high power objective (x400), we evaluated IDCI in 10 regions of surgical specimen including the most invasive slice and averaged the number of S-100-protein positive cells. The patients were classified into two groups according to IDCI (>20 or <20 S-100-protein positive cells per high power field: high or low IDCI, respectively). The degree of IDCI varied from 0 to 67 (average 12.3). The 203 patients were separated into 65 with high IDCI and 138 with low IDCI. The depth of invasion was significantly more superficial in patients with high IDCI (p<0.05), and the disease was at an earlier clinical stage (p<0.01) than in those with low IDCI. Five-year survival rates after curative resection were 67% and 39% in patients with high and low IDCI, respectively (p<0.01). Multivariate analysis showed that IDCI was an independent prognostic factor (RR=1.6, p=0.02), next to nodal involvement, depth of invasion and clinical stage. Being closely related to clinical features, the prognosis of patients with esophageal cancer may be estimated by monitoring IDCI.

Clinical significance of circulating tumor cells in blood by molecular detection and tumor markers in esophageal cancer.

BACKGROUND: The clinical significance of circulating tumor cells in the blood during surgery has not been elucidated in esophageal squamous cell carcinoma (ESCC). We evaluated the relationship between circulating tumor cells and clinicopathologic findings, compared with that of serum squamous cell carcinoma (SCC) antigen and carcinoembryonic antigen (CEA), in ESCC. METHODS: Blood samples from 54 consecutive patients were obtained from the peripheral artery and the superior vena cava at three points in time: immediately before surgery, and before and after tumor resection. CEA-specific reverse transcriptase-polymerase chain reaction (RT-PCR), which can quantify circulating tumor cells in blood, was performed. The preoperative values of serum SCC antigen and CEA were also obtained for all patients. RESULTS: CEA messenger RNA (CEA mRNA) was detected in the blood of 31 out of 54 patients (57.4%). CEA mRNA positivity was detected most frequently after tumor resection and correlated with nodal status and stage grouping. The incidence of total recurrence and blood-borne recurrence was significantly greater in patients with CEA mRNA positivity than in those with CEA mRNA negativity (P =.036 and.0026, respectively). Preoperative serum levels of SCC antigen and CEA did not correlate with clinicopathologic findings and tumor recurrence. CONCLUSIONS: CEA mRNA detected by RT-PCR was more predictive of tumor recurrence than serum tumor markers. Effective adjuvant therapy is recommended for patients with CEA mRNA positive expression.

Perioperative detection of circulating cancer cells in patients with colorectal hepatic metastases.

BACKGROUND/AIMS: Surgical resection is the most effective therapy for metastatic colorectal cancer to the liver. However, the selection criteria for patients who may benefit from partial hepatectomy are not fully defined. The aim of this study was to evaluate the usefulness of perioperative molecular detection of circulating cancer cells in predicting clinical outcome in patients with colorectal metastatic liver cancer. METHODOLOGY: Blood samples were obtained perioperatively from the portal vein, peripheral artery, and superior vena cava in 16 consecutive patients with colorectal liver metastases who have undergone partial hepatic resection. We analyzed circulating cancer cells using a carcinoembryonic antigen-specific nested reverse transcriptase-polymerase chain reaction. RESULTS: Positive carcinoembryonic antigen-mRNA expression was detected in 7 (43.8%) of 16 patients. Six (85.7%) of those 7 patients had hematogenous rerecurrences during the 1- to 3-year follow-up period. None of 9 negative-patients showed re-recurrence (p < 0.01). Among the 9 of 16 patients receiving postoperative adjuvant chemotherapy, no clear effect was noted regarding re-recurrence in the positive carcinoembryonic antigen-mRNA patients. CONCLUSIONS: These results suggest that the molecular detection of circulating cancer cells at the time of surgery for colorectal liver metastases could be one measure of high-risk patients for hematogenous re-recurrence after partial hepatic resection.

Biologic and imaging diagnosis of lymph node metastasis in esophageal carcinoma.

BACKGROUND AND OBJECTIVES: Few reports have described the combined use of biologic and imaging techniques in the diagnosis of lymph node metastasis. We prospectively evaluated lymph node metastasis diagnosed by biologic and imaging means in patients with esophageal carcinoma. METHODS: Preoperative ultrasound and endoscopic ultrasound (EUS) examination were performed in 80 patients. Biopsy specimens were immunohistochemically examined using cyclin D1 (CD1) and desmoglein 1 (DG1) antibodies, and tumors were classified into three grades. RESULTS: The sensitivity, specificity, and accuracy values of ultrasound examination were 88.2, 58.6, and 77.5%, respectively. The incidence of nodal involvement was 0% (0/10) in patients with grade 1 tumors, 57.1% (16/28) in those with grade 2 tumors, and 83.3% (35/42) in those with grade 3 tumors. Of the 57 patients with lymph node metastasis determined sonographically, 50 had grade 2 or 3 tumors that were histologically confirmed. The remaining seven patients with grade 1 tumors did not have involved nodes. Of the 23 patients without lymph node metastasis according to ultrasound examination, the incidence of lymph node metastasis in patients with grade 1, 2, and 3 tumors was 0, 16.7, and 50.0%, respectively. CONCLUSIONS: When used together, imaging and molecular procedures may offer improved identification of lymph node metastasis in patients with squamous cell carcinoma of the esophagus.

Smad4 and transforming growth factor beta1 expression in patients with squamous cell carcinoma of the esophagus.

PURPOSE: The members of the Smad family play key rolesin regulating gene expression in the transforming growth factor (TGF)-beta1 signaling pathways. Activation of Smads causes their translocation from the cytoplasm to the nucleus, where they function as transcription factors. The present study analyzed the expression and clinicopathological significance of Smad4 and TGF-beta1 in squamous cell carcinoma of the esophagus. EXPERIMENTAL DESIGN: Immunohistochemistry was used to investigate the expression of Smad4 and TGF-beta1 proteins in 258 patients with squamous cell carcinoma of the esophagus. The relationship between expression of these proteins and clinicopathological factors was analyzed, and the usefulness of Smad4 in disease prognosis was evaluated in relation to TGF-beta1 expression. RESULTS: Smad4 expression was preserved in 32.2% of tumors, and TGF-beta1 expression was identified in 42.6% of tumors. Patients with preserved expression of Smad4 had a higher rate of early-stage carcinoma (P < 0.01) and fewer lymph node metastases (P < 0.01) than those with reduced Smad4 expression. The expression of TGF-beta1 was not associated with any of the clinicopathological factors. Postoperative survival analysis indicated that patients with a tumor in which Smad4 expression was reduced had worse clinical outcomes than those with preserved expression (P = 0.01). In patients with TGF-beta1-negative tumors, the survival rate was significantly higher in patients with a preserved level of Smad4 expression than in those with reduced Smad4 expression (P = 0.02). However, according to multivariate analysis, Smad4 expression could not be used as an independent prognostic factor. CONCLUSIONS: Although Smad4 expression could not be used as a prognostic factor, its expression reflected tumor progression such as tumor depth and lymph node metastasis.

Lymph node micrometastasis and lymphatic mapping determined by reverse transcriptase-polymerase chain reaction in pN0 gastric carcinoma.

BACKGROUND: The purposes of this study were to examine lymph node micrometastasis (LMM) by reverse transcriptase-polymerase chain reaction (RT-PCR) method, and clarify the initial nodes involved by metastatic disease according to tumor location. METHODS: We examined 312 lymph nodes obtained from 50 patients with node-negative gastric carcinoma. RT-PCR and immunohistochemistry were performed. The clinical characteristics of LMM were investigated, and the map of LMM was made according to tumor location. RESULTS: The number of patients and LMM detected by RT-PCR was 14 and 17 and by immunohistochemistry was 7 and 8, respectively. RT-PCR was a more sensitive method than immunohistochemistry. LMM by RT-PCR correlated with depth of tumor invasion and lymphatic invasion. Regarding pT1 tumor, 9 patients with LMM had tumors that were of the macroscopically depressed type and 2 cm or more in diameter. According to the lymphatic map, right pericardial lymph nodes and lymph nodes along the lesser curvature were the initial nodes involved in the upper third of the stomach. Right pericardial lymph nodes, lymph nodes along the lesser curvature, and infrapyloric nodes were more important initial metastatic sites in the middle third of the stomach, and lymph nodes along the lesser curvature and infrapyloric nodes in the lower third. CONCLUSIONS: We demonstrated the relationship between LMM and clinicopathologic factors, especially in pT1 tumor. The mapping of LMM may prove useful for selecting the optimal treatment.

Biological properties of biopsy specimens are useful for predicting lymph node micrometastasis in esophageal carcinoma.

BACKGROUND: Some studies have reported on lymph node micrometastasis (MM) by RT-PCR. We attempted to predict MM by biological means using preoperative biopsy specimens. MATERIALS AND METHODS: Lymph nodes from 60 patients with esophageal carcinoma were examined by routine histological examination and CEA-specific RT-PCR. The biopsy specimens were immunohistochemically examined using p53, cyclin D1 (CD1) and desmoglein 1 (DG1) antibodies. RESULTS: Of 659 lymph nodes, 53 (8.0%) nodes were positive according to histological examination and 158 (24.0%) had MM by RT-PCR. The percentage of patients with lymph node metastasis according to histological examination and RT-PCR was 65.0% and 81.7%, respectively. CD1 and DG1 expression correlated with MM, whereas p53 expression did not. MM was frequently detected in the tumors with CD1-positive, DG1-negative or reduced expression. CONCLUSION: The expression of CD1 and DG1 in biopsy specimens may offer useful information on lymph node metastasis, including MM in esophageal carcinoma.

Paraaortic lymph node micrometastasis and tumor cell microinvolvement in advanced gastric carcinoma.

BACKGROUND: Paraaortic lymph node dissection in advanced gastric carcinoma is controversial. The purpose of this study was to investigate the incidence and significance of micrometastasis (MM) or tumor cell microinvolvement (TCM) in these critical lymph nodes.METHODS: A total of 2339 lymph nodes, including 390 paraaortic nodes, obtained from 47 patients with advanced gastric carcinoma were examined immunohistochemically, using cytokeratin antibody.RESULTS: Lymph node metastasis was found in 95 of the 390 paraaortic nodes of 14 patients by routine histological examination. MM or TCM was immunohistochemically detected in 45 of the 295 negative paraaortic lymph nodes from 15 of 33 patients (MM, n = 5; TCM, n = 10). The 5-year-survival rate in the paraaortic node-negative group and cytokeratin-positive group was significantly higher that that of the hematoxilin and eosin-positive group. The total number of lymph node metastases by hematoxylin and eosin staining and the pathological lymph node compartments, by cytokeratin-positive nodes, were prognostic factors by multivariate analysis.CONCLUSIONS: We demonstrated a high rate of MM or TCM in the paraaortic lymph nodes and suggest that such harbored metastases are related to the prognosis of patients with advanced gastric carcinoma. On the basis of this study, a multi-institutional study should be considered.