Reactive swelling of the regional lymph nodes in patients with stage II colorectal cancer as a prognostic factor.

The aim of this study was to examine the relationship between reactive swelling of regional lymph node (LN) and survival in colorectal carcinoma. We retrospectively studied 170 patients with surgically resected colorectal carcinoma histopathologically diagnosed as stage II (pStage II). These patients were classified into two groups:a) a "reactive LN swelling" group (clinically diagnosed as positive for LN metastasis, but pathologically negative) and b) a "no LN swelling" group. Survival analyses of the two groups showed that overall survival (OS) and disease-specific survival (DSS) were longer in the reactive LN swelling group than in the no LN swelling group in patients with total colorectal and right-sided colon cancer. Multivariate analyses revealed that reactive LN swelling was an independent prognostic factor in OS and DSS in patients with right-sided colon cancer. Reactive swelling of regional LN is regarded as an expression of local immune responses, which could explain the present results.

Phase I study of gemcitabine, cisplatin, and S-1 combination therapy for patients with untreated advanced biliary tract cancer.

BACKGROUND: To develop a triplet regimen containing gemcitabine, cisplatin, and S-1 (GPS), we assessed the recommended dose for patients with untreated advanced biliary tract cancer in this phase I study. METHODS: Dose-limiting toxicities (DLTs) were evaluated for the following two dose levels: gemcitabine (1000 mg/m(2) for level 1 and 1200 mg/m(2) for level 2 on day 1), cisplatin (30 mg/m(2) fixed dose on day 1), and S-1 (40-60 mg/day fixed dose twice a day for 7 days), every 2 weeks until progression. DLTs for each level were evaluated in six or more patients during the first two cycles. RESULTS: A total of 18 patients were enrolled and 16 patients were evaluated. DLTs at level 1 were observed in two of 10 patients. At level 2, a DLT was observed in one of six patients. The main grade 3 or 4 treatment-related adverse events were neutropenia and leukopenia, and a few non-hematological toxicities were observed. Among 14 patients with measurable lesions, the best response rate was 50%. CONCLUSIONS: GPS with a relative dose intensity corresponding to 90% of the standard gemcitabine plus cisplatin regimen could be administered safely, and showed preliminary antitumor activity. Survival benefits will be studied subsequently.

Glasgow Prognostic Score predicts poor prognosis among advanced biliary tract cancer patients with good performance status.

Advanced cancer patients with good performance status (PS) sometimes show poor prognosis despite receiving some chemotherapies. We evaluated prognosis of chemo-naïve advanced biliary tract cancer (ABTC) patients with good PS by Glasgow Prognostic Score (GPS). Sixty-two patients with Eastern Cooperative Oncology Group PS 0 or 1 were retrospectively analyzed, using multivariate Cox regression. GPS was defined with serum levels of two parameters, albumin >3.5 g/dl and C-reactive protein <1.0 mg/dl (both as 0, either as 1, and neither as 2). PS 0 (n = 32) and 1 (n = 30) patients had similar survival (P = 0.98). The median overall survival (OS) was 17.0 months for GPS 0 (n = 19), 14.2 months for GPS 1 (n = 17), and 6.4 months for GPS 2 (n = 26). GPS 2 had significantly shorter OS than GPS 0 (P = 0.002) or 1 (P = 0.033). Multivariate analysis identified two independent prognostic factors: GPS (hazard ratio 0.60, 95 % confidence interval 0.40-0.90, P = 0.012) and liver metastasis (hazard ratio 0.43, 95 % CI 0.20-0.90, P = 0.026). GPS was useful for chemo-naïve ABTC patients with good PS.

Bevacizumab in combination with irinotecan, 5-fluorouracil, and leucovorin (FOLFIRI) in patients with metastatic colorectal cancer who were previously treated with oxaliplatin-containing regimens: a multicenter observational cohort study (TCTG 2nd-BV study).

The efficacy of bevacizumab combined with infusional 5-fluorouracil/leucovorin (5-FU/LV) plus irinotecan (FOLFIRI) as the second-line treatment for metastatic colorectal cancer (mCRC) has not been fully clarified, although bevacizumab combined with infusional 5-FU/LV plus oxaliplatin (FOLFOX) in the second-line setting has demonstrated a survival benefit. We investigated the efficacy of bevacizumab plus FOLFIRI in mCRC patients who failed oxaliplatin-containing regimens without bevacizumab. Patients who received bevacizumab plus FOLFIRI or bevacizumab plus FOLFOX as second-line chemotherapy between July 2007 and March 2008 were registered (trial registration: UMIN000001547). Patient background data and progression-free survival (PFS), overall survival (OS), response, and bevacizumab-related adverse events were prospectively collected every 6 months. A total of 195 patients were enrolled from 26 institutions. Among them, 115 patients received bevacizumab plus FOLFIRI after failure of oxaliplatin and fluoropyrimidine (FOLFIRI+BV after OX/FU group), and 45 patients received bevacizumab plus FOLFOX after failure of irinotecan and fluoropyrimidine (FOLFOX+BV after IRI/FU group). Median PFS was 8.3 months (95% confidence interval [CI], 6.7-9.9) for the FOLFIRI+BV after OX/FU group and 7.8 months (95% CI, 5.8-9.7) for the FOLFOX+BV after IRI/FU group. Median OS was 21.6 months (95% CI, 17.6-25.6) and 16.5 months (95% CI, 11.8-21.2), respectively. Overall response rates were 25 and 29%, respectively. The most common grade≥3 bevacizumab-related adverse events were hypertension (5.0%) and bleeding (3.8%). FOLFIRI+BV after OX/FU showed comparable efficacy to FOLFOX+BV after IRI/FU.

Apoptosis and inhibition of the phosphatidylinositol 3-kinase/Akt signaling pathway in the anti-proliferative actions of dehydroepiandrosterone.

BACKGROUND: Dehydroepiandrosterone (DHEA) is an endogenous steroid that is synthesized mainly in the adrenal cortex; it is found in plasma as the sulfate-conjugated form (DHEA-S). Pharmacological doses of DHEA exhibit anti-proliferative effects on malignant cell lines and some tumors in experimental animals. The purpose of this study was to evaluate the effect of these steroids on proliferation in human cancer cell lines. METHODS: HepG2 and HT-29 cell lines were treated with DHEA or DHEA-S at 0-200 microM for 24 h or at 100 microM for 8-72 h, and then effects on cell growth, and the cell cycle and on apoptosis, were evaluated by 3-[4,5-dimethylthiazol]-2yl-2,5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry, respectively. Also, the effect of DHEA on phosphatidylinositol 3-kinase (PI3K)/Akt signaling was investigated in HepG2 cells by Western blotting. RESULTS: The growth of HepG2 and HT-29 cells was significantly inhibited by DHEA, in a dose- and time-dependent manner. This inhibition was greater in HepG2 than in HT-29 cells. Accumulation at G0/G1 phase in both cell lines was observed with DHEA treatment. However, apoptosis increased significantly only in HepG2 cells. In contrast, DHEA-S exhibited much weaker growth inhibitory and cytostatic effects on both cell lines, and apoptosis was not detected. In HepG2 cells treated with DHEA, apoptosis was associated with markedly reduced Akt phosphorylation (Thr308 and Ser473), suggesting that DHEA inhibited the PI3K/Akt signaling to induce apoptosis in these cells. CONCLUSIONS: These results suggest that the induction of apoptosis through the inhibition of the PI3K/Akt signaling pathway is one of the anti-proliferative mechanisms of DHEA in certain tumors, but that DHEA also promotes cell-cycle arrest without the induction of apoptosis.

Simultaneous determination of dehydroepiandrosterone and its 7-oxygenated metabolites in human serum by high-resolution gas chromatography--mass spectrometry.

A highly sensitive and specific method has been developed for the simultaneous measurement of free (unconjugated) or sulfate-conjugated forms of dehydroepiandrosterone (DHEA), 7alpha-hydroxy-DHEA (7alpha-OH-DHEA), 7beta-hydroxy-DHEA (7beta-OH-DHEA), and 7-oxo-DHEA (7-oxo-DHEA) in human serum. This method is based upon a stable isotope-dilution technique by gas chromatography-selected-ion monitoring mass spectrometry. Free steroids were extracted from serum with an organic solvent and the sulfate-conjugated steroids remained in aqueous phase. Free steroids were purified by solid-phase extraction, while sulfate-conjugated steroids were hydrolyzed by sulfatase and deconjugated steroids were purified by solid-phase extractions. The extracts were treated with O-methylhydroxylamine hydrochloride and were subsequently dimethylisopropylsilylated. The resulting methyloxime-dimethylisopropylsilyl (MO-DMIPS) ether derivatives were quantified by gas chromatography-selected-ion monitoring mass spectrometry in a high-resolution mode. The detection limits of MO-DMIPS ether derivatives of DHEA, 7alpha-OH-DHEA, 7beta-OH-DHEA and 7-oxo-DHEA were 1.0, 0.5, 0.5 and 2.0pg, respectively. Coefficients of variation between samples ranged from 10.6 to 22.9% for free 7-oxygenated DHEA to less than 10% for DHEA and sulfate-conjugated 7-oxygenated DHEA. The concentrations of these steroids were measured in 18 sera samples from healthy volunteers (9 males and 9 females; aged 23-78 years). Free DHEA, 7alpha-OH-DHEA, 7beta-OH-DHEA and 7-oxo-DHEA levels ranged between 0.21-3.55, 0.001-0.194, 0.003-0.481, and 0.000-0.077ng/ml, respectively, and the sulfate-conjugated steroid levels of these metabolites ranged between 253-4681, 0.082-3.001, 0.008-0.903, and 0.107-0.803ng/ml, respectively. The free DHEA-related steroid concentrations were much lower than those previously measured by RIA and low-resolution GC-MS. The present method made it possible to determine simultaneously serum DHEA-related steroid levels with sufficient sensitivity and accuracy.

Anti-proliferative action of endogenous dehydroepiandrosterone metabolites on human cancer cell lines.

Dehydroepiandrosterone (DHEA) is a naturally occurring steroid synthesized in the adrenal cortex, gonads, brain, and gastrointestinal tract, and it is known to have chemopreventive and anti-proliferative actions on tumors. These effects are considered to be induced by the inhibition of glucose-6-phosphate dehydrogenase (G6PD) and/or HMG-CoA reductase (HMGR) activities. The present study was undertaken to investigate whether endogenous DHEA metabolites, i.e. DHEA-sulfate, 7-oxygenated DHEA derivatives, androsterone, epiandrosterone, and etiocholanolone, have anti-proliferative effects on cancer cells and to clarify which enzyme, G6PD or HMGR, is responsible for growth inhibition. Growth of Hep G2, Caco-2, and HT-29 cells, evaluated by 3-[4,5-dimethylthiazol]-2yl-2,5-diphenyl tetrazolium bromide (MTT) and bromodeoxyuridine incorporation assays, was time- and dose-dependently inhibited by addition of all DHEA-related steroids we tested. In particular, the growth inhibition due to etiocholanolone was considerably greater than that caused by DHEA in all cell lines. The suppression of growth of the incubated steroids was not correlated with the inhibition of G6PD (r=-0.031, n=9, NS) or HMGR (r=0.219, n=9, NS) activities. The addition of deoxyribonucleosides or mevalonolactone to the medium did not overcome the inhibition of growth induced by DHEA or etiocholanolone, while growth suppression by DHEA was partially prevented by the addition of ribonucleosides. These results demonstrate that endogenous DHEA metabolites also have an anti-proliferative action that is not induced by inhibiting G6PD or HMGR activity alone. These non-androgenic DHEA metabolites may serve as chemopreventive or anti-proliferative therapies.

Selective inhibition of CYP27A1 and of chenodeoxycholic acid synthesis in cholestatic hamster liver.

The aim of this study was to explore the regulation of serum cholic acid (CA)/chenodeoxycholic acid (CDCA) ratio in cholestatic hamster induced by ligation of the common bile duct for 48 h. The serum concentration of total bile acids and CA/CDCA ratio were significantly elevated, and the serum proportion of unconjugated bile acids to total bile acids was reduced in the cholestatic hamster similar to that in patients with obstructive jaundice. The hepatic CA/CDCA ratio increased from 3.6 to 11.0 (P<0.05) along with a 2.9-fold elevation in CA concentration (P<0.05) while the CDCA level remained unchanged. The hepatic mRNA and protein level as well as microsomal activity of the cholesterol 7alpha-hydroxylase, 7alpha-hydroxy-4-cholesten-3-one 12alpha-hydroxylase and 5beta-cholestane-3alpha,7alpha,12alpha-triol 25-hydroxylase were not significantly affected in cholestatic hamsters. In contrast, the mitochondrial activity and enzyme mass of the sterol 27-hydroxylase were significantly reduced, while its mRNA levels remained normal in bile duct-ligated hamster. In conclusion, bile acid biosynthetic pathway via mitochondrial sterol 27-hydroxylase was preferentially inhibited in bile duct-ligated hamsters. The suppression of CYP27A1 is, at least in part, responsible for the relative decreased production of CDCA and increased CA/CDCA ratio in the liver, bile and serum of cholestatic hamsters.