An Active 32-kDa Cathepsin L Is Secreted Directly from HT 1080 Fibrosarcoma Cells and Not via Lysosomal Exocytosis.

Cathepsin L [EC 3.4.22.15] is secreted via lysosomal exocytosis by several types of cancer cells, including prostate and breast cancer cells. We previously reported that human cultured fibrosarcoma (HT 1080) cells secrete cathepsin L into the medium; this secreted cathepsin is 10-times more active than intracellular cathepsin. This increased activity was attributed to the presence of a 32-kDa cathepsin L in the medium. The aim of this study was to examine how this active 32-kDa cathepsin L is secreted into the medium. To this end, we compared the secreted active 32-kDa cathepsin L with lysosomal cathepsin L by using a novel gelatin zymography technique that employs leupeptin. We also examined the glycosylation and phosphorylation status of the proteins by using the enzymes endoglycosidase H [EC 3.2.1.96] and alkaline phosphatase [EC 3.1.3.1]. Strong active bands corresponding to the 32-kDa and 34-kDa cathepsin L forms were detected in the medium and lysosomes, respectively. The cell extract exhibited strong active bands for both forms. Moreover, both forms were adsorbed onto a concanavalin A-agarose column. The core protein domain of both forms had the same molecular mass of 30 kDa. The 32-kDa cathepsin L was phosphorylated, while the 34-kDa lysosomal form was dephosphorylated, perhaps because of the lysosomal marker enzyme, acid phosphatase. These results suggest that the active 32-kDa form does not enter the lysosomes. In conclusion, our results indicate that the active 32-kDa cathepsin L is secreted directly from the HT 1080 cells and not via lysosomal exocytosis.

Comparison of intratumoral heterogeneity of HER2 expression between primary tumor and multiple organ metastases in gastric cancer: Clinicopathological study of three autopsy cases and one resected case.

Intratumoral heterogeneity of HER2 expression in the metastatic foci of HER2-positive advanced gastric cancer remains unclear. In this study, we compared HER2 expression between primary and metastatic tumors in HER2-positive three autopsied cases and one resected case with multiple organ metastases by immunohistochemistry (IHC) and dual color in situ hybridization (DISH). All four cases judged positive (IHC3+) at the primary tumor tissues showed varying HER2 gene amplification (GA) status. One homogeneously HER2-positive autopsied case (Case 1) and one intratumorally heterogeneous positive resected case (Case 2) with high GA showed a homogeneous positive staining pattern in all the metastatic foci. One heterogeneously HER2-positive autopsied case (Case 3) with low GA showed a partially heterogeneous HER2 staining pattern in all the metastatic foci. In contrast, one heterogeneously HER2-positive autopsied case (Case 4) with equivocal GA showed a completely heterogeneous HER2 staining pattern in the metastatic foci. These results indicate that HER2-positive gastric cancers with low to high GA at the primary tumor show substantially homogeneous HER2 overexpression in the metastatic foci, whereas HER2-positive gastric cancers with equivocal GA expressed HER2 heterogeneously within the metastatic tumor, suggesting that metastatic foci of the latter HER2-positive cases would be potentially resistant to trastuzumab.

Long-term outcomes and prognostic factors of patients with advanced gastric cancer treated with S-1 plus cisplatin combination chemotherapy as a first-line treatment.

BACKGROUND: The long-term outcomes of advanced gastric cancer (AGC) patients treated with S-1 plus cisplatin (SP) combination chemotherapy remain unclear. Therefore, we sought to evaluate these outcomes to identify the prognostic factors affecting patient survival. METHODS: We retrospectively analyzed 153 AGC patients treated with SP at a single institution between January 2005 and July 2011. RESULTS: Median overall survival (OS) was 15.0 months [95 % confidence interval (CI), 12.5-17.9 months]. Three independent prognostic factors affecting poor survival were identified: performance status (PS) ≥ 1 [hazard ratio (HR) = 2.39, 95 % CI, 1.58-3.62); >1 metastatic site (HR = 1.57, 95 % CI, 1.10-2.26], and elevated alkaline phosphatase levels (HR = 1.70, 95 % CI, 1.16-2.49). A simple prognostic index was generated using three risk groups: good (no risk factor), moderate (one or two risk factors), and poor (three risk factors). The median OS for good-, moderate-, and poor-risk groups was 28.6, 14.8, and 7.3 months, respectively (log-rank test; P < 0.0001). Among the twelve 3-year survivors, 9 (75 %) had a PS of 0 and 8 (67 %) had only one metastatic site. CONCLUSIONS: Three prognostic factors were identified in AGC patients treated with SP. Using a simple prognostic index, the patients were divided into three risk groups, in which the survival differences were markedly significant, suggesting that patients with good PS and only one metastatic site may have a higher chance of long-term survival than those with poor PS and multiple metastatic sites.

CA19-9 level as a prognostic and predictive factor of bevacizumab efficacy in metastatic colorectal cancer patients undergoing oxaliplatin-based chemotherapy.

PURPOSE: The prognostic and predictive values of carbohydrate antigen 19-9 (CA19-9) levels in metastatic colorectal cancer (mCRC) remain unclear. We reviewed all mCRC patients at a single institution to evaluate the relationship between CA19-9 levels and survival. METHODS: Two hundred and fifty-two patients underwent first-line chemotherapy using oxaliplatin-based regimens between April 2005 and December 2009. The relationship between baseline CA19-9 levels and survival was analyzed. Moreover, we evaluated the relationship between baseline CA19-9 levels and clinicopathological factors. RESULTS: One hundred and fifty patients had elevated baseline CA19-9 levels (elevated group), and 79 patients had normal baseline CA19-9 (normal group) levels. Both KRAS and BRAF mutation rates were higher in the elevated group than in the normal group. Elevated CA19-9 level was a poor prognostic factor compared with normal CA19-9 levels (P = 0.0021). In the elevated group, the median survival time with bevacizumab was significantly longer with bevacizumab than without it (median OS, 27.8 vs. 15.3 months, P = 0.0019). However, the median survival time was not different with or without bevacizumab in the normal group (median OS, 36.5 vs. 38.0 months, P = 0.9515). CONCLUSION: Our results suggest that baseline CA19-9 level is an independent prognostic factor in mCRC patients, and it correlated with the KRAS/BRAF mutation status. Bevacizumab exhibits clinical activity only for high CA19-9 levels in mCRC.

A retrospective comparison of S-1 plus cisplatin and capecitabine plus cisplatin for patients with advanced or recurrent gastric cancer.

BACKGROUND: Based on the results of the SPIRITS trial, combination chemotherapy of S-1 plus cisplatin (SP) is now considered the standard treatment for patients with advanced gastric cancer (AGC) in Japan. On the other hand, several non-Japanese studies have shown the efficacy of capecitabine plus cisplatin (XP), which has been used as the reference arm in recent global studies of AGC. METHODS: We retrospectively compared the efficacy and safety of SP and XP in first-line treatment for patients with AGC. RESULTS: From August 2006 to November 2008, 26 AGC patients received XP in the context of 2 global trials (AVAGAST and ToGA), and 50 patients received SP during the same period. The objective response rate was 43.2 % in the SP group and 50 % in the XP group, with no significant difference (p = 0.62). There were also no significant differences in progression-free survival (median 5.8 vs. 5.2 months; p = 0.91) and overall survival (median 13.8 vs. 13.5 months; p = 0.97) between the SP and XP groups. The frequencies of hematological toxicities of grade 3 or more and non-hematological toxicities were not significantly different between the 2 groups. Although grade 1 or 2 hand-foot syndrome was more common in the XP group, no patients experienced grade 3 or more. CONCLUSIONS: Although the retrospective nature of this study and the small number of patients is a major limitation, SP and XP were associated with similar efficacy and safety in patients with AGC.

Prognosis of patients with advanced gastric cancer by HER2 status and trastuzumab treatment.

BACKGROUND: The purpose of this study was to evaluate the impact of human epidermal growth factor receptor 2 (HER2) status and trastuzumab treatment on the prognosis of patients with advanced gastric cancer (AGC). METHODS: We retrospectively analyzed 364 AGC patients who received systemic chemotherapy. To evaluate the impact of trastuzumab exposure during any type of chemotherapy, our analysis used time-varying covariates to avoid a possible lead-time bias. RESULTS: Among the 364 patients, 58 (15.9 %) were HER2-positive. The median overall survival of the HER2-positive patients treated with trastuzumab (n = 43) was significantly longer than that of the HER2-negative patients [n = 306; 24.7 vs. 13.9 months, with an adjusted hazard ratio (HR) of 0.58; 95 % confidence interval (CI), 0.36-0.95; P = 0.03]. Notably, 22 patients continued with trastuzumab beyond the date of progression. By contrast, the HER2-positive patients not treated with trastuzumab (n = 15) showed survival similar to that of the HER2-negative patients (13.5 vs. 13.9 months, with an adjusted HR of 1.04; 95 % CI, 0.52-2.11; P = 0.91). According to the multivariate analysis, exposure to trastuzumab was independently associated with a better prognosis (HR 0.56; 95 % CI; 0.33-0.93; P = 0.026). CONCLUSIONS: Recent HER2-positive AGC patients have a better prognosis than HER2-negative patients, particularly when treated with trastuzumab.

Fluoropyrimidine plus cisplatin for patients with advanced or recurrent gastric cancer with peritoneal metastasis.

BACKGROUND: There are few data on the efficacy of combination chemotherapy with a fluoropyrimidine plus cisplatin for patients with advanced or recurrent gastric cancer (AGC) complicated by peritoneal metastasis, especially massive ascites. METHODS: We retrospectively evaluated the efficacy and safety of a fluoropyrimidine (S-1 or capecitabine) plus cisplatin as first-line chemotherapy in 120 patients with AGC and peritoneal metastasis. RESULTS: Ascites was detected in 50 patients, with 11 patients having massive ascites. Median progression-free survival (PFS) and overall survival (OS) of all patients was 6.1 and 15.9 months, respectively. The PFS and OS were shorter in patients with massive ascites (n = 11; 3.7 and 9.5 months) compared with patients with small or moderate ascites (n = 39; 5.8 and 13.5 months) or patients without ascites (n = 70; 6.9 and 18.1 months). The objective response in terms of ascites was similar whether ascites was massive (4 of 11 patients; 36.4%) or small or moderate (16 of 39 patients; 41%). The frequencies of grade 3 or higher toxicity or treatment discontinuation due to toxicity are relatively similar across ascites groups. CONCLUSIONS: Fluoropyrimidine plus cisplatin appears to be tolerated in selected patients with peritoneal metastasis.

Reporting patient characteristics and stratification factors in randomized trials of systemic chemotherapy for advanced gastric cancer.

BACKGROUND: There is no consensus on which patient characteristics are the most suitable to report or to be used as stratification factors in clinical trials for advanced gastric cancer (AGC), to our knowledge. METHODS: We conducted a comprehensive review of published randomized trials for AGC to examine the patient characteristics that were reported. RESULTS: Among the 67 analyzed trials, age, gender, performance status, proportion of patients with measurable disease, and previous gastrectomy were frequently reported (>69%). Histology, number of disease sites, and adjuvant treatment were reported in less than 50% of trials. Although the reporting of second-line chemotherapy has increased in recent trials, it remains at less than 50%. Notably, recent trials have tended to include patients with better performance status and less locally advanced disease, with Asian trials more frequently including patients with more diffuse histology and less locally advanced disease or liver metastasis than non-Asian trials. Stratification was conducted in approximately 60% of the trials, using quite variable stratifying factors. CONCLUSION: Inconsistency exists in the reporting of patient characteristics, the characteristics themselves, and the use of stratification factors in clinical trials for AGC. A consensus set of important patient characteristics and strata may be necessary to conduct and interpret quality randomized studies.

Prolonged survival of patients with metastatic colorectal cancer following first-line oxaliplatin-based chemotherapy with molecular targeting agents and curative surgery.

BACKGROUND: The ability of molecular targeting agents to improve overall survival (OS) in metastatic colorectal cancer (MCRC) patients who underwent oxaliplatin-based chemotherapy remains controversial. METHODS: We retrospectively analyzed 331 patients with MCRC who underwent first-line oxaliplatin-based chemotherapy. Treatment outcomes were compared between patients who started chemotherapy from April 2005 to March 2007 (cohort A; n = 157) and those who started it from April 2007 to March 2009 (cohort B; n = 174). To evaluate the impact of exposure to agents, we applied time-varying covariate analysis to avoid possible lead-time bias. RESULTS: Median OS of cohorts A and B was 21.3 and 28.6 months, respectively (HR 0.66, 95% CI 0.50-0.87, p = 0.003). Exposure to bevacizumab (25 vs. 76%), anti-epidermal growth factor receptor (EGFR) (18 vs. 33%) or curative surgery after chemotherapy (4 vs. 10%) was significantly higher in cohort B. According to a multivariate Cox model with exposure to each agent or treatment as a time-varying covariate, hazard ratios of death were 0.71 (95% CI, 0.51-0.96; p = 0.03) for bevacizumab, 0.62 (95% CI, 0.40-0.89; p = 0.01) for anti-EGFR and 0.22 (95% CI, 0.06-0.57; p = 0.004) for surgery. CONCLUSIONS: Increased exposure to molecular targeting agents or surgery after chemotherapy appears to contribute to an improvement in OS in recent patients with MCRC who have undergone oxaliplatin-based chemotherapy.

A retrospective comparison of docetaxel and paclitaxel for patients with advanced or recurrent esophageal cancer who previously received platinum-based chemotherapy.

OBJECTIVE: To retrospectively compare docetaxel (DTX) with paclitaxel (PTX) with regard to efficacy and safety in advanced or recurrent esophageal cancer patients who previously received platinum-based chemotherapy. METHODS: We retrospectively analyzed 124 advanced or recurrent esophageal cancer patients who had received platinum-based chemotherapy and then received DTX or PTX from April 2006 to November 2010. RESULTS: Eighty-six patients (69.4%) received DTX and 38 patients (30.6%) received PTX monotherapy. Due to toxicity, dose reduction was needed in 36.0 and 27.8% of patients and treatment was discontinued in 10.5 and 2.6% of patients receiving DTX and PTX, respectively. The objective response (25.7 vs. 10.3%, p = 0.03) and disease control rates (60.0 vs. 34.6%, p = 0.01) were higher in the PTX group than in the DTX group, respectively. There were no significant differences in median progression-free survival (2.1 vs. 3.5 months) and overall survival (6.1 vs. 7.2 months) between the DTX and PTX groups, respectively. Grade 3-4 neutropenia (48.8 vs. 21.1%, p = 0.003) and febrile neutropenia (20.9 vs. 5.3%, p = 0.029) were more frequent in the DTX patients than in the PTX patients, respectively. CONCLUSION: Although the efficacy of DTX and PTX for advanced or recurrent esophageal cancer patients after platinum-based chemotherapy was not significantly different in terms of survival, PTX was a more feasible treatment. PTX provided similar efficacy to DTX with less febrile neutropenia.

Video-rate bioluminescence imaging of matrix metalloproteinase-2 secreted from a migrating cell.

BACKGROUND: Matrix metalloproteinase-2 (MMP-2) plays an important role in cancer progression and metastasis. MMP-2 is secreted as a pro-enzyme, which is activated by the membrane-bound proteins, and the polarized distribution of secretory and the membrane-associated MMP-2 has been investigated. However, the real-time visualizations of both MMP-2 secretion from the front edge of a migration cell and its distribution on the cell surface have not been reported. METHODOLOGY/PRINCIPAL FINDINGS: The method of video-rate bioluminescence imaging was applied to visualize exocytosis of MMP-2 from a living cell using Gaussia luciferase (GLase) as a reporter. The luminescence signals of GLase were detected by a high speed electron-multiplying charge-coupled device camera (EM-CCD camera) with a time resolution within 500 ms per image. The fusion protein of MMP-2 to GLase was expressed in a HeLa cell and exocytosis of MMP-2 was detected in a few seconds along the leading edge of a migrating HeLa cell. The membrane-associated MMP-2 was observed at the specific sites on the bottom side of the cells, suggesting that the sites of MMP-2 secretion are different from that of MMP-2 binding. CONCLUSIONS: We were the first to successfully demonstrate secretory dynamics of MMP-2 and the specific sites for polarized distribution of MMP-2 on the cell surface. The video-rate bioluminescence imaging using GLase is a useful method to investigate distribution and dynamics of secreted proteins on the whole surface of polarized cells in real time.

Docetaxel plus 5-fluorouracil and cisplatin (DCF) induction chemotherapy for locally advanced borderline-resectable T4 esophageal cancer.

BACKGROUND: This study aimed to evaluate the efficacy of docetaxel plus 5-fluorouracil and cisplatin (DCF) induction chemotherapy for locally advanced borderline-resectable T4 esophageal cancer. PATIENTS AND METHODS: We retrospectively analyzed data regarding thirty patients with borderline-resectable T4 tumor who received either DCF or cisplatin plus 5-fluorouracil (FP) as induction chemotherapy. RESULTS: The overall response rate was significantly better for the DCF group than the FP group. In the DCF group, 6/16 patients achieved a grade 2 histological post-chemotherapeutic effect after treatment, compared to 1/14 in FP group. Except for myelotoxicity, no other significant differences in toxicity were observed during induction chemotherapy between groups. The DCF regimen did not result in increased postoperative complications compared to the FP regimen. Postoperative recurrence or distant metastasis was observed in 7/10 of FP patients and 5/12 of DCF patients. CONCLUSION: DCF induction chemotherapy may be an option for conversion therapy of initially unresectable, locally advanced esophageal cancer.

FOLFOX plus cetuximab for a patient with metastatic colorectal cancer with icterus due to multiple liver metastases.

The prognosis of patients with advanced colorectal cancer with icterus is dismally poor, and adequate chemotherapy for these patients has not been established yet. A 59-year-old male with fatigue, anorexia and icterus with serum total bilirubin 9.7 mg/dL was referred to our institution. He was diagnosed with advanced sigmoid colon cancer with multiple liver metastases. A biopsy specimen of the primary tumor showed well-differentiated adenocarcinoma without KRAS mutation. Since biliary drainage was impossible due to diffuse liver metastases, we initiated combination chemotherapy with 5-fluorouracil, Leucovorin, oxaliplatin (modified FOLFOX6) and cetuximab. The doses of 5-fluorouracil and oxaliplatin were reduced, but cetuximab was administered at the standard dosage. After 3 courses of chemotherapy, total bilirubin dropped to 0.8 mg/dL. No significant toxicity other than grade-2 skin toxicity and neuropathy was observed, and the patient has continued chemotherapy on an outpatient basis. Combination chemotherapy with mFOLFOX6 plus cetuximab was effective and feasible in this case of metastatic colon cancer with icterus due to diffuse liver metastasis.

A case of heavily pretreated rectal cancer with disseminated intravascular coagulation that improved following reintroduction of FOLFOX plus bevacizumab.

Disseminated intravascular coagulation (DIC) is a complication that may be experienced by patients with solid tumors. The prognosis of solid tumors with DIC is much poorer than those without DIC. Although treatment of the underlying disease is critical for improvement of DIC, the efficacy and safety of chemotherapy in patients with DIC associated with colorectal cancer are not clear. A 50-year-old man with advanced rectal cancer and multiple liver metastases experienced DIC during third-line treatment with cetuximab plus irinotecan, following 5-fluorouracil, leucovorin, oxaliplatin (FOLFOX) plus bevacizumab and 5-fluorouracil, leucovorin, and irinotecan (FOLFIRI) plus bevacizumab. Combination chemotherapy consisting of FOLFOX plus bevacizumab was reintroduced. Although platelet and fresh-frozen plasma transfusions were required daily before chemotherapy, the patient's laboratory values improved after two cycles of chemotherapy, without severe toxicity. The patient was discharged, and FOLFOX plus bevacizumab has been continued on an outpatient basis without sign of recurrence of DIC as of December 2010 (4 months after initiation of chemotherapy). This case suggests that reintroduction of combination chemotherapy with FOLFOX plus bevacizumab is effective and feasible in patients with colorectal cancer with DIC and that chemotherapy may be a treatment option for such patients.

Retrospective analysis of cetuximab monotherapy for patients with irinotecan-intolerant metastatic colorectal cancer.

BACKGROUND: The efficacy and safety of cetuximab for irinotecan-intolerant patients has not yet been evaluated in detail. METHODS: We retrospectively analyzed the efficacy and safety of cetuximab monotherapy for patients with metastatic colorectal cancer (MCRC) that was intolerant to irinotecan. RESULTS: Among 105 patients who received cetuximab-containing chemotherapy until March 2010, 22 patients were treated with cetuximab monotherapy due to irinotecan intolerance. Cetuximab was given at the approved dosage to all patients. The performance status was 2 or 3 in 17 patients (77%). All but 1 patient had wild-type KRAS tumors. The causes of irinotecan intolerance were icterus (n = 9; 41%; median serum total bilirubin, 6.3 mg/dl), symptomatic peritoneal metastasis or obstruction (n = 8; 36%), and thrombocytopenia (n = 1; 5%). Four patients (18%) refused irinotecan due to previous irinotecan-associated toxicity. Two patients achieved a partial response with an apparent drop of serum bilirubin, for a response rate of 9.1%. The median progression-free survival and overall survival were 1.6 and 3.5 months, respectively. No grade 3 or 4 adverse events or treatment-related deaths were experienced. CONCLUSION: Cetuximab monotherapy for irinotecan-intolerant MCRC is feasible. However, the overall efficacy was modest in the present cohort, despite the fact that most of the patients had wild-type KRAS tumors; further effective therapies should be evaluated to improve the prognosis of this patient population.

Association of fluoropyrimidines, platinum agents, taxanes, and irinotecan in any line of chemotherapy with survival in patients with advanced gastric cancer.

BACKGROUND: Although fluoropyrimidines, platinum agents, taxanes, and irinotecan are used in the treatment of advanced gastric cancer (AGC), it remains unclear whether these agents in any line of chemotherapy are associated with overall survival (OS) in these patients. METHODS: We retrospectively analyzed 704 patients with AGC. To avoid possible lead-time bias, we applied time-varying covariate analysis for chemotherapy with four agents in any line. RESULTS: Median OS was 12.3 months. The frequency of exposure to each agent class during all lines of treatment was 92.6% for FU (5-fluorouracil or oral fluoropyrimidine), 48.2% for platinum agents, 65.1% for taxanes, and 39.1% for irinotecan. According to a multivariate Cox model with exposure to each agent class as a time-varying covariate, the hazard ratios (HRs) of death were 0.41 (95% confidence interval [CI], 0.27-0.57; p < 0.001) for FU, 0.71 (95% CI, 0.58-0.84; p < 0.001) for platinum agents, 0.51 (95% CI, 0.41-0.63; p < 0.001) for taxanes, and 0.53 (95% CI, 0.43-0.65; p < 0.001) for irinotecan. Although other agents were used in 18.6% of the patients, they did not affect survival. CONCLUSIONS: Each of the four agent classes (FU, platinum agents, taxanes, and irinotecan) appears to be independently associated with improved OS in patients with AGC regardless of timing. This result suggests the importance of developing strategies which make these active agents available to all patients with AGC to prolong OS.

Lapatinib plus trastuzumab for a patient with heavily pre-treated gastric cancer that progressed after trastuzumab.

A 57-year-old female with advanced gastric cancer was referred to our hospital. She underwent neoadjuvant chemotherapy with S-1 plus cisplatin followed by curative gastrectomy. Weekly paclitaxel and combination chemotherapy with irinotecan plus cisplatin were administered for lymph node recurrence, but the tumor progressed. Since the human epidermal growth factor receptor 2 status of her gastric cancer specimen was strongly positive, docetaxel plus trastuzumab was administered for three cycles. However, the lymph node metastasis appeared to enlarge and abdominal pain worsened. Therefore, combination chemotherapy with lapatinib and weekly trastuzumab was initiated. A computed tomographic scan after 3 months of treatment showed stable disease. Although dose reduction of lapatinib was necessary due to Grade 3 diarrhea, the patient has continued this treatment on an outpatient basis without signs of disease progression for 8 months after initiation. In this case, trastuzumab plus lapatinib resulted in durable stable disease, despite the appearance of progression during prior chemotherapy with trastuzumab.

Meta-analysis of neutropenia or leukopenia as a prognostic factor in patients with malignant disease undergoing chemotherapy.

PURPOSE: We performed a systematic review and meta-analysis to determine the impact of neutropenia or leukopenia experienced during chemotherapy on survival. METHODS: Eligible studies included prospective or retrospective analyses that evaluated neutropenia or leukopenia as a prognostic factor for overall survival or disease-free survival. Statistical analyses were conducted to calculate a summary hazard ratio and 95% confidence interval (CI) using random-effects or fixed-effects models based on the heterogeneity of the included studies. RESULTS: Thirteen trials were selected for the meta-analysis, with a total of 9,528 patients. The hazard ratio of death was 0.69 (95% CI, 0.64-0.75) for patients with higher-grade neutropenia or leukopenia compared to patients with lower-grade or lack of cytopenia. Our analysis was also stratified by statistical method (any statistical method to decrease lead-time bias; time-varying analysis or landmark analysis), but no differences were observed. CONCLUSIONS: Our results indicate that neutropenia or leukopenia experienced during chemotherapy is associated with improved survival in patients with advanced cancer or hematological malignancies undergoing chemotherapy. Future prospective analyses designed to investigate the potential impact of chemotherapy dose adjustment coupled with monitoring of neutropenia or leukopenia on survival are warranted.

Randomized phase II study comparing dose escalated weekly paclitaxel vs. standard dose weekly paclitaxel for patients with previously treated advanced gastric cancer.

Weekly paclitaxel is an effective and widely used regimen for patients with advanced gastric cancer, with main dose-limiting toxicities of neutropenia and neurotoxicity. Neutropenia during weekly paclitaxel administration was reported to be associated with better survival. The aim of this study is to evaluate prospectively whether dosing adjustments based on the occurrence of neutropenia may improve chemotherapy efficacy. A total of 90 patients will be randomized to receive either a standard dose of weekly paclitaxel (80 mg/m(2)) or an escalated dose of weekly paclitaxel (80 mg/m(2) initially followed by 100 and 120 mg/m(2) unless severe toxicity is observed). The primary endpoint is overall survival. Secondary endpoints include progression-free survival, response rate, disease control rate and adverse events.

Significance of 32-kDa cathepsin L secreted from cancer cells.

It is recognized that many cancer cells secrete cathepsin L to degrade the components of extracellular matrices and basement membranes, thus promoting tumor invasion and metastasis. However, very little information is available concerning the secreted forms of cathepsin L and their possible role in human cancer. We initially demonstrated that approximately 10-fold higher mature cathepsin L activity was secreted in a medium of human fibrosarcoma (HT 1080) cells, compared with their intracellular activity. A 32-kDa major-activity band, together with a 41-kDa faint-activity band, was detected in the medium by our newly developed gelatin zymography. The two forms were further confirmed to be cathepsin L by immunoblot analysis. Both were apparently secreted directly from the cells, as neither was affected when the cells were cultured in the presence of various kinds of proteinase inhibitors. Human tumor necrosis factor-alpha (TNF-alpha) stimulated not only the production of the 32-kDa cathepsin L, but also its secretion. Moreover, the 32-kDa cathepsin L activities in 3 colon and 2 lung cancer tissues were significantly higher than in normal tissues. Based on the foregoing, there are good reasons to speculate that the 32-kDa cathepsin L found in HT 1080 cell medium is involved in cancer invasion and metastasis.