[Long-term survival and effective treatment of synchronous liver metastases in a case of primary gastric choriocarcinoma by using hepatic arterial infusion chemotherapy, irinotecan, and cisplatin].

A 62-year-old man was admitted to our hospital because of appetite loss. Computed tomography(CT)revealed thickness of the gastric wall, multiple liver tumors, and lung nodes. Upper gastrointestinal endoscopy revealed an easy bleeding type 2 tumor at the gastric antrum. We performed distal gastrectomy to control bleeding from the gastric tumor. Histological findings from the gastric lesion indicated primary gastric choriocarcinoma(PCG). Combination chemotherapy using hepatic arterial infusion chemotherapy for synchronous liver metastases and S-1 was administered for 5 months after the operation. CT revealed that the liver metastases decreased remarkably. On the other hand, lung metastases increased. Irinotecan and cisplatin were administered. Liver metastases did not increase, as observed using imaging studies. The patient died 17 months after the operation for cachexia. PCG is a highly aggressive tumor that is often associated with liver metastasis. It is important to control liver metastasis from PCG.

Diffusely infiltrative squamous cell carcinoma of the esophagus presenting as a case with diagnostic difficulty.

We report a rare case of diffusely infiltrative squamous cell carcinoma of the esophagus. The patient, a 68-year-old man, was admitted to our hospital for investigation of dysphagia. Esophagography and upper gastrointestinal endoscopy revealed esophageal stenosis and gastric cancer. As a biopsy from the esophagus revealed no sign of malignancy, he underwent only distal gastrectomy. After 3 months, the stenosis became worse. Again, biopsy from a gastrointestinal endoscopy showed no malignancy, but endobronchial ultrasound-guided transbronchial needle aspiration revealed squamous cell carcinoma of the esophagus. Despite various treatments, the patient died of disease progression 20 months after its onset. Autopsy revealed diffusely infiltrative squamous cell carcinoma of the esophagus, which is a rare malignancy with few case reports documented.

[A case of recurrent rectal cancer with idiopathic thrombocytopenic purpura showing rapid progression, suggesting disseminated intravascular coagulopathy].

A 64-year-old female with idiopathic thrombocytopenic purpura(ITP)was admitted to our hospital under the diagnosis of rectal cancer. Intersphincteric resection and splenectomy were performed after high-dose gamma globulin therapy. Thirteen months after the surgery, she suffered from a local recurrence and groin and pelvic lymph node metastases. Radiotherapy was planned before curative resection. During radiation, she complained of severe back pain and high fever with severe thrombocytopenia, and was admitted to our hospital. The examinations revealed disseminated intravascular coagulopathy(DIC), probably induced by multiple bone and hepatic metastases. Although anti-DIC therapy and chemotherapy with FOLFIRI were performed, thrombocytopenia did not improve, and she died of cancer progression about 2 months after admission. We report a case ofDIC induced by cancer progression with ITP. Since thrombocytopenia may be induced by either DIC or ITP, selecting a treatment for such a patient is difficult. We report the present case in detail and discuss findings from the literature.

COX2 expression predicts resistance to chemoradiotherapy in esophageal squamous cell carcinoma.

PURPOSE: The overexpression of cyclooxygenase (COX)2 is correlated with carcinogenesis, tumor progression, and prognosis, and increased COX2 expression is correlated with radiation resistance. However, no correlation between the COX2 expression and resistance to chemoradiotherapy for esophageal squamous cell carcinoma has been characterized. The purpose of the present study was to evaluate whether COX2 expression is an indicator of resistance to chemoradiotherapy in esophageal squamous cell carcinoma and the feasibility of COX2 as a biomarker for CRT. METHODS: Fifty-eight patients who were diagnosed with esophageal squamous cell carcinoma from biopsy samples were enrolled in the present series. All patients underwent concurrent chemoradiotherapy in a neoadjuvant setting, followed by radical esophagectomy. COX2 expression was evaluated by immunohistochemical staining and statistically compared with the histopathologic findings in surgically resected specimens. RESULTS: The rate of responders was 87% for weak expression of COX2, 62% for moderate expression, and 30% for strong expression, and there was a close correlation between COX2 expression and the response rate (Kendall's τb = 0.396, P = 0.001). In the univariate analysis, negative or weak expression of COX2 was found to correlate significantly with CRT response (odds ratio, 6.296; 95% confidence interval (CI), 1.58-25.096; P = 0.010). In the multivariate analysis, weak expression of COX2 (30% or less) was found to be an independent prognostic factor (odds ratio, 6.534; 95% CI, 1.535-27.803; P = 0.011). CONCLUSIONS: The COX2 expression predicts resistance to chemoradiotherapy in esophageal squamous cell carcinoma, and it also is a feasible biomarker for evaluating the CRT response.

Correlation between gp96 expression and the surgical outcome in patients with esophageal squamous cell carcinoma.

BACKGROUND: Heat-shock protein gp96 plays an important role in antitumor immunoreactions. Gp96 has a close relationship with antitumor immunity. This study evaluated the correlation between gp96 expression and the prognosis in esophageal squamous cell carcinoma. METHODS: Seventy-eight patients with primarily resected esophageal squamous cell carcinoma were enrolled onto this study, and gp96 expression was evaluated by immunohistochemical staining. The association of clinicopathological factors and patients' survival was calculated by univariate (log rank test) and multivariate (Cox proportional hazard regression method) analyses. RESULTS: Fifty-seven (73%) of 78 cases were gp96 positive, and 21 were negative (27%). The survival of patients with gp96-negative disease was significantly shorter (5-year survival, 22.9 months) than with gp96-positive disease (45.8 months; P = 0.049), and the multivariate analysis showed that gp96 negativity is an independent risk factor for poor survival (hazard ratio, 2.577; P = 0.040). Gp96-negative cases had more metastatic lymph nodes than did negative cases, especially in T1 cases (4.8 in gp96-negative cases vs. 0.84 in gp96-positive cases; P = 0.064) CONCLUSIONS: The downregulation of gp96 expression is closely correlated with poor survival in esophageal squamous cell carcinoma.

Postoperative lymphocyte percentage influences the long-term disease-free survival following a resection for colorectal carcinoma.

OBJECTIVE: The aim of this study is to examine the relationship between postoperative laboratory parameters of inflammation and the disease-free survival in patients undergoing resection for colorectal cancer. METHODS: Six hundred seventy-five consecutive patients who underwent an elective resection for primary colorectal cancer from October 1999 to March 2004 were included in this study. We examined the associations between cancer recurrence and white blood cell count, lymphocyte percentage, neutrophil percentage and C-reactive protein. RESULTS: Lymphocyte percentage on postoperative days 3 and 7 was significantly higher in patients without recurrence than in those with recurrence. Lymphocyte percentage on postoperative day 7 differed the most between the two groups. On postoperative day 7, Stage II patients with lymphocyte percentage >15% had significantly longer survival compared with the patients with lymphocyte percentage ≤ 15%. A multivariate analysis showed lymphocyte percentage ≤ 15% on postoperative day 7 to be an independent prognostic factor, along with lymph node metastases and serosal invasion. Logistic regression analysis showed that blood loss (>250 ml) and postoperative complications were significant independent predictors of lymphocyte percentage ≤ 15% on postoperative day 7. CONCLUSIONS: Lymphocyte percentage ≤ 15% on postoperative day 7 is an independent prognostic factor for the patients undergoing a resection for colorectal cancer.

Oncologic outcome of intersphincteric resection for very low rectal cancer.

BACKGROUND: In 2000 we launched a prospective program of intersphincteric resection (ISR) for very low rectal cancer. In this study we compared the oncologic outcome of patients who underwent ISR with the outcome of patients who underwent abdominoperineal resection (APR). METHODS: The data of 202 patients with very low rectal cancer who underwent curative ISR (n = 132) or curative APR (n = 70) between 1995 and 2006 were analyzed. Patients were divided into ISR and APR groups. Survival and local recurrence were investigated in both groups. RESULTS: The median follow-up was 40 months in the ISR group and 57 months in the APR group. The 5-year local relapse-free survival rate was 83% in the ISR group and 80% in the APR group (p = 0.364), and the 5-year disease-free survival rate was 69% in the ISR group and 63% in the APR group (p = 0.714). CONCLUSIONS: For very low rectal cancers, ISR appears to be oncologically acceptable and can reduce the number of APRs.

Combined effects of p53 gene therapy and leptomycin B in human esophageal squamous cell carcinoma.

BACKGROUND: p53 gene therapy has been examined in several clinical trials, however, the results of those trials have mostly been unsatisfactory due to the low efficacy of this therapy. Leptomycin B (LMB) is an antibiotic originally isolated from Streptomyces that has the ability to inhibit the export of proteins containing a nuclear export signal from the nucleus to the cytoplasm. Currently, it has been shown that p53 protein has a nuclear export signal. In this study, we assessed whether LMB augments the transduced p53 gene effect. METHODS: Antiproliferative effect of LMB was assessed in human esophageal squamous cancer cell lines. Accumulation of p53 protein into the nucleus by LMB was observed by fluorescence microscopy. The combined effect of p53 and LMB was evaluated in in vitro experiments. RESULTS: LMB induced cell death in a dose-dependent manner and p53 drastically accumulated in the nucleus after LMB treatment. The combinatory treatment of p53 gene and LMB significantly increases the efficiency compared to either agent alone. CONCLUSIONS: Our findings suggest that LMB has a potent ability to augment the effect of the tumor suppressor p53 in esophageal squamous cancer cell lines and that it is a promising component in p53 gene therapy.

Role of histone deacetylase inhibitor in adenovirus-mediated p53 gene therapy in esophageal cancer.

BACKGROUND: Recently, the use of histone deacetylase inhibitors (HDACI) with gene therapy has been shown to improve the effect of this therapy. The effectiveness of one of the novel HDACIs, FK228, was examined in adenovirus-mediated p53 gene therapy of esophageal squamous cell carcinoma (ESCC). MATERIALS AND METHODS: The expression levels of coxsackie and adenovirus receptor (CAR) in ESCC patients were examined immunohistochemically. CAR induction by FK228 in ESCC cells was analyzed by real-time PCR and Western blotting. The efficiencies of adenoviral transduction treated with FK228 were determined using AV1.0CMV-betagal. The acetylation of p53 protein was detected by Western blotting. RESULTS: CAR expression was reduced in some tumor specimens compared to that in normal specimens. CAR expression was increased by FK228 in both in vitro and in vivo experiments. FK228 improved the efficiency of adenovirus infection. Acetylated p53 protein was increased in a dose- and a time-dependent manner. CONCLUSION: Our findings suggest that FK228 has a potent ability to augment the effect of adenovirus-mediated p53 gene therapy in ESCC cells.

Prognostic significance of peritoneal washing cytology in patients with potentially resectable gastric cancer.

BACKGROUND/AIMS: The prognostic value of cytological examination of intraoperative washings in potentially resectable gastric cancer is controversial. METHODOLOGY: Between February 1993 and August 2001, clinicopathological features and surgical outcome of 26 consecutive patients with gastric cancer with positive cytological findings of peritoneal washings without peritoneal dissemination were retrospectively analyzed. RESULTS: The overall 1, 2, 3-year survival rates for 26 patients were 69%, 35%, and 0%, respectively. The median survival was 17.5 months. The median survival of patients with curative resection (n=16) and non-curative resection (n=10) was 19 months and 12.5 months, respectively. There was no significant difference in survival between curative resection and non-curative resection (p=0.10). Recurrent disease frequently occurred as peritoneal dissemination (69%). No patient survived for more than 34 months. CONCLUSIONS: Aggressive surgical resections do not provide any survival benefit for gastric cancer with positive cytological findings of peritoneal washings even in the absence of peritoneal dissemination.

Combination of direct intratumoral administration of dendritic cells and irradiation induces strong systemic antitumor effect mediated by GRP94/gp96 against squamous cell carcinoma in mice.

We tested a new therapeutic modality for head and neck and esophageal cancers, a combination of direct intratumoral (i.t.) administration of dendritic cells (DCs) and radiation therapy (RT) in mouse squamous cell carcinoma (SCC). We also evaluated the functions of gp96, which can enhance systemic antitumor activity, and the mechanism of the abscopal effect. Mouse SCC cells (1 x 10(5)), SCCVII, were inoculated into the left femur of C3H/He mice subcutaneously, and also similarly inoculated into chest subcutaneous tissue. Only the left femur tumor was exposed to 4 or 10 Gy of ionizing radiation, and then 1 x 10(6) DCs i.t. was injected only into the femur tumor. Following this procedure, tumor volumes of the femur and chest were measured. We evaluated whether gp96 could enhance the antitumor effect. With DCs i.t. and RT, tumor growth was markedly suppressed. Tumor growth of non-treated tumors were also suppressed, indicating that the combination therapy of DCs and RT evoked systemic antitumor activity. In vitro, the enhancement of gp96 expression was strongly detected by immunostaining after irradiation, DCs with gp96 induced strong cytotoxic activity in vitro, and tumor growth inhibition was observed by direct i.t. injection of gp96. A combination of DCs i.t. and RT can induce a strong antitumor effect not only against treated local tumor but also against non-treated distant tumor, indicating that this treatment can evoke a strong systemic antitumor effect. Gp96 is thought to be one of the target molecules to explain the abscopal effect.

Tumor suppressor Prdx1 is a prognostic factor in esophageal squamous cell carcinoma patients.

Peroxiredoxins (Prdxs) are a family of antioxidant enzymes that are also known as scavengers of peroxide in mammalian cells. Some reports have shown that the overexpression of Prdx1, which is one of the peroxiredoxins that is a ubiquitously expressed protein, was related to a poor prognosis in several types of human cancers. In this study, we investigated the expression levels of Prdx1 in esophageal squamous cell carcinoma by immunohistochemistry, and the correlation between the Prdx1 expression and the clinical status was elucidated. Immunohistochemical staining was performed in 114 samples which were collected from surgical esophageal cancer specimens. Cytoplasmic staining of Prdx1 was evaluated based on the following scoring criteria: Grade I, negative or weak staining; Grade II, moderate staining; and Grade III, strong staining. The percentage of patients with a Grade I expression of Prx1 was 20% (23 of 114), 44% had Grade II (50 of 114), and 36% had Grade III (41 of 114). The Prdx1 immunoreactivity showed an inverse significant correlation with T-category (P<0.0001), lymph node metastasis (P=0.048), and stage (P=0.001). In addition, the patients with tumors exhibiting a reduced Prdx1 expression had shorter overall survival (P=0.022) in comparison to the patients with tumors which had a higher Prdx1 expression. Currently, Prdx1 has been shown to act as a tumor suppressor. Our results provide strong evidence that the reduced Prdx1 expression is an important factor in esophageal squamous cancer progression and could serve as a useful prognostic marker.

Gene expression profiling induced by histone deacetylase inhibitor, FK228, in human esophageal squamous cancer cells.

Histone deacetylase inhibitors (HDACIs) are promising therapeutic agents with the potential for regulating cell cycle, differentiation and apoptosis in cancer cells. HDACI activity is associated with selective transcriptional regulation and altering gene expression. However, the exact mechanisms leading to the antitumor effect of HDACIs are not fully understood. FK228, one of the powerful HDACIs, strongly inhibited cell growth of T.Tn and TE2 cells and induced apoptosis. Therefore, comprehensive analysis of the changes in gene expression in human esophageal cancer cell lines by the HDACI FK228 was carried out by microarray analysis. This analysis was used to clarify the expression profiles of genes after exposure to FK228. Of the 4,608 genes analyzed, 93 genes in T.Tn and 65 genes in TE2 were up- or down-regulated 2-fold or more at least at one time point during FK228 exposure and they were classified into four clusters based on their expression patterns. Among them, 15 genes were contained in both cell lines and their expression patterns were similar. Except p21, Prdx1 (reported by us) and IGFBP3, the behaviour/expression of 12 highly responsive genes has still not been reported in esophageal cancer cells. These observations of the expression patterns of functionally classified genes provided insights into the mechanism of the antitumor effect of FK228 in esophageal squamous carcinoma.

[Two cases treated with trastuzumab as primary chemotherapy].

We report two cases treated with primary chemotherapy containing trastuzumab with a review of some important papers. The first patient was a 43-year-old female. A 33-mm left breast invasive ductal carcinoma (ER (-), PgR (-), HER2 3+(IHC) ) with several lymph node metastases in the Ax, Ic and Sc was found. After primary chemotherapy with 6 courses of EC and 4 courses of weekly paclitaxel + trastuzumab, the efficacy for the local tumor was judged as PR. However, brain metastases appeared, so the operation was canceled. Brain metastases were then treated by gamma-knife three times, but systemic chemotherapy was not administered. Eight months later, carcinomatous meningitis appeared. Intrathecal chemotherapy with MTX+Ara-C was started, but the patient died after 20 months from the beginning of the treatment. Local efficacy was judged as CR. The second patient was a 41-year-old female. A 39-mm right breast invasive ductal carcinoma (ER (-), PgR (-), HER2 3+(IHC) ) with two lymph node metastases in the Ax was found. After primary chemotherapy with 6 courses of FEC and 4 courses of weekly paclitaxel + trastuzumab, the efficacy was judged as PR. The operation was scheduled, but he patient wished to continue chemotherapy for cosmetic reasons. Later, because of mild tumor regrowth, we used 2 courses of vinorelbine in combination with trastuzumab. The tumor grew more, so Bp + Ax was done. The woman is alive at this writing with no recurrence.

Histone deacetylase inhibitor FK228 activates tumor suppressor Prdx1 with apoptosis induction in esophageal cancer cells.

PURPOSE: The histone deacetylase inhibitor FK228 shows strong activity as a potent antitumor drug but its precise mechanism is still obscure. The purpose of this study is to reveal the effect of FK228 on gene expression in the cell and to determine the mechanism of the antitumor activity of FK228 for further clinical applications. EXPERIMENTAL DESIGN AND RESULTS: Microarray analysis was applied to verify the gene expression profiles of 4,608 genes after FK228 treatment using human esophageal squamous cell cancer cell lines T.Tn and TE2. Among them, peroxiredoxin 1 (Prdx1), a member of the peroxiredoxin family of antioxidant enzymes having cell growth suppression activity, as well as p21(WAF1), were significantly activated by FK288. In addition, FK228 strongly inhibited the cell growth of T.Tn and TE2 by the induction of apoptosis. Further, chromatin immunoprecipitation analysis revealed that FK228 induced the accumulation of acetylated histones H3 and H4 in Prdx1 promoter, including the Sp1-binding site. In mouse xenograft models of T.Tn and TE2 cells, FK228 injection resulted in significant tumor regression as well as activated Prdx1 expression in tumor tissues. Prdx1 suppression by RNA interference hindered the antitumor effect of FK228. CONCLUSION: Our results indicate that the antitumor effect of FK228 in esophageal cancer cells is shown at least in part through Prdx1 activation by modulating acetylation of histones in the promoter, resulting in tumor growth inhibition with apoptosis induction.

miR-203 inhibits the migration and invasion of esophageal squamous cell carcinoma by regulating LASP1.

The expression of microRNA-203 (miR-203) in esophageal squamous cell carcinoma (ESCC) tissues is remarkably lower than that in non­ESCC tissues. We investigated how miR-203 could influence the development of ESCC cells. Our analyses revealed that miR-203 inhibited the migration and invasion of ESCC cells. Genome-wide gene expression data and target site inhibition assays showed that miR-203 appears to directly regulate LIM and SH3 protein 1 (LASP1). The knockdown of LASP1 resulted in inhibition of the migration and invasion of ESCC cells. Our results suggest that miR-203 and its target LASP1, may be associated with the progression of ESCC. In clinical ESCC specimens, the expression levels of miR-203, which were lower compared to those in normal tissues, were inversely correlated with the mRNA expression levels of LASP1. Moreover, we found that there was a significant correlation between the expression levels of miR-203 and the relapse­free survival. The identification of a cancer network regulated by miR-203 could provide new insights into the potential mechanisms of the progression of ESCC.

Identification of novel molecular targets regulated by tumor suppressive miR-375 induced by histone acetylation in esophageal squamous cell carcinoma.

The aim of this study was to determine whether histone acetylation regulates tumor suppressive microRNAs (miRNAs) in esophageal squamous cell carcinoma (ESCC) and to identify genes which are regulated by these miRNAs. We identified a miRNA that was highly upregulated in an ESCC cell line by cyclic hydroxamic acid-containing peptide 31 (CHAP31), one of the histone deacetylase inhibitors (HDACIs), using a miRNA array analysis. miR-375 was strongly upregulated by CHAP31 treatment in an ESCC cell line. The expression levels of the most upregulated miRNA, miR-375 were analyzed by quantitative real-time PCR in human ESCC specimens. The tumor suppressive function of miR-375 was revealed by restoration of miR-375 in ESCC cell lines. We performed a microarray analysis to identify target genes of miR-375. The mRNA and protein expression levels of these genes were verified in ESCC clinical specimens. LDHB and AEG-1/MTDH were detected as miR­375-targeted genes. The restoration of miR-375 suppressed the expression of LDHB and AEG-1/MTDH. The ESCC clinical specimens exhibited a high level of LDHB expression at both the mRNA and protein levels. A loss-of-function assay using a siRNA analysis was performed to examine the oncogenic function of the gene. Knockdown of LDHB by RNAi showed a tumor suppressive function in the ESCC cells. The correlation between gene expression and clinicopathological features was investigated by immunohistochemistry for 94 cases of ESCC. The positive staining of LDHB correlated significantly with lymph node metastasis and tumor stage. It also had a tendency to be associated with a poor prognosis. Our results indicate that HDACIs upregulate miRNAs, at least some of which act as tumor suppressors. LDHB, which is regulated by the tumor suppressive miR-375, may therefore act as an oncogene in ESCC.

RIG-I helicase-independent pathway in sendai virus-activated dendritic cells is critical for preventing lung metastasis of AT6.3 prostate cancer.

We recently demonstrated highly efficient antitumor immunity against dermal tumors of B16F10 murine melanoma with the use of dendritic cells (DCs) activated by replication-competent, as well as nontransmissible-type, recombinant Sendai viruses (rSeV), and proposed a new concept, "immunostimulatory virotherapy," for cancer immunotherapy. However, there has been little information on the efficacies of this method: 1) in more clinically relevant situations including metastatic diseases, 2) on other tumor types and other animal species, and 3) on the related molecular/cellular mechanisms. In this study, therefore, we investigated the efficacy of vaccinating DCs activated by fusion gene-deleted nontransmissible rSeV on a rat model of lung metastasis using a highly malignant subline of Dunning R-3327 prostate cancer, AT6.3. rSeV/dF-green fluorescent protein (GFP)-activated bone marrow-derived DCs (rSeV/dF-GFP-DC), consistent with results previously observed in murine DCs. Vaccination of rSeV/dF-GFP-DC was highly effective at preventing lung metastasis after intravenous loading of R-3327 tumor cells, compared with the effects observed with immature DCs or lipopolysaccharide-activated DCs. Interestingly, neither CTL activity nor DC trafficking showed any apparent difference among groups. Notably, rSeV/dF-DCs expressing a dominant-negative mutant of retinoic acid-inducible gene I (RIG-I) (rSeV/dF-RIGIC-DC), an RNA helicase that recognizes the rSeV genome for inducing type I interferons, largely lost the expression of proinflammatory cytokines without any impairment of antitumor activity. These results indicate the essential role of RIG-I-independent signaling on antimetastatic effect induced by rSeV-activated DCs and may provide important insights to DC-based immunotherapy for advanced malignancies.

Development of immunostimulatory virotherapy using non-transmissible Sendai virus-activated dendritic cells.

Dendritic cell (DC)-based immunotherapy has been clinically evaluated, however, still requires modification to improve its outcomes. We previously demonstrated that DCs activated by replication competent recombinant Sendai virus (SeV) showed dramatic efficacy over that seen in use of current DC vaccine for immunotherapy against malignancies; however, application of replication-deficient vector is more relevant in clinical setting. We here show that F-gene-deleted non-transmissible Sendai virus (SeV/dF)-activated DCs (DCs/SeV/dF) has strong antitumor effects against murine SCCVII tumor, that was well-known as a less immunogenic cell line. SeV/dF shows high transfection efficiency to DCs and leads them to upregulate costimulatory molecules. Intratumoral injection of DCs/SeV/dF resulted in a marked and representative inhibition of the tumor, even when the tumors were well-vascularized. This is the first demonstration that non-transmissible SeV vector, SeV/dF, could be a DC-activator; DC/SeV/dF-based cancer immunotherapy may, therefore, warrant further investigation.