A Phase II Study of S-1 Monotherapy as a First-line Combination Therapy of S-1 Plus Cisplatin as a Second-line Therapy, and Weekly Paclitaxel Monotherapy as a Third-line Therapy in Patients with Advanced Gastric Carcinoma: A Second Report

Background We have previousy reported on a Phase II study of S-1 monotherapy as a first line, combination therapy of S-1 plus cisplatin as a second line, and weekly paclitaxel monotherapy as a third line therapy in patients with advanced gastric carcinomas. The median survival time (MST) of patients over the whole course of treatment was not previously calculated because 12 out of 19 patients had not yet succumbed. Since then, we have calculated the MST for this study and herein report our findings. Patients and Methods Between 2002 and 2005, 19 patients were enrolled in this study. Chemotherapy consisted of either 60 mg/m 2 of S-1 for 4 weeks at 6-week intervals, a combination of 60 mg/m 2 S-1 for 3 weeks and 60 mg/m 2 cisplatin on day 8 at 5-week intervals, or 60 mg/m 2 paclitaxel at days 1, 8, and 15, at 4-week intervals. The regimens were repeated until the occurrence of unacceptable toxicities, disease progression, or patient noncompliance. The primary end point was the overall survival. Results The median survival time was 774 days. The response rates were 33.3% (3/9), 12.5% (1/8), and 0% (0/4) after the first, second, and third line chemotherapies, respectively. The major adverse hematological toxicity was leukopenia, which reached grades 3–4 in all lines of chemotherapy investigated. In addition, the major adverse non-hematological toxicity was anorexia, which reached grade 3–4 in second line chemotherapy, and no deaths were attributable to the adverse effects of the drugs. Conclusion This sequential therapy was an effective treatment for advanced gastric cancer with acceptable toxic side-effects. We considered this therapy to be effective because of the smooth transition to the next regimen

Discovery of a novel quinoxalinhydrazide with a broad-spectrum anticancer activity

Previously, we discovered a novel class of salicylhydrazide compounds with remarkable activity in hormone-dependent and -independent human cancer cells. We then designed and synthesized numerous analogues. Among these analogues, a quinoxalinhydrazide compound, SC144, exhibited desirable physicochemical and drug-like properties and therefore was selected for further preclinical investigation. In the present study, we evaluated the in vitro activity of SC144 in a range of drug-sensitive and -resistant cancer cell lines as well as its in vivo efficacy in MDA-MB-435 and HT29 mice xenograft models. The broad-spectrum cytotoxicity of SC144 is especially highlighted by its potency in ovarian cancer cells resistant to cisplatin, breast-cancer cells resistant to doxorubicin, and colon cancer cells resistant to oxaliplatin. Furthermore, its activity was independent of p53, HER-2, estrogen and androgen receptor expressions. We also examined the effect of SC144 on cell cycle progression and apoptosis in select cell lines. Considering its cytotoxicity profile in a variety of in vitro and in vivo cancer models as well as its effects on cell cycle regulation and apoptosis, SC144 appears to represent a promising agent for further clinical development

Phase II Study of S-1 Monotherapy as a First-line, Combination Therapy of S-1 plus Cisplatin as a Second-line, and Weekly Paclitaxel Monotherapy as a Third-line Therapy in Patients with Advanced Gastric Carcinoma

Background We conducted a pilot phase II study to evaluate the efficacy and safety of S-1 as a first-line, S-1 plus cisplatin as a second-line, and weekly paclitaxel as a third-line therapy for advanced gastric cancer. Patients and Methods Between 2002 and 2005, 19 patients were enrolled in this study. Chemotherapy consisted of either 60 mg/m 2 of S-1 for 4 weeks at 6 weeks interval, a combination of 60 mg/m 2 S-1 for 3 weeks and 60 mg/m 2 cisplatin on day 8 at 5 weeks interval, or 60 mg/m 2 paclitaxel at day 1, 8, 15, at 4 weeks interval. The regimen was repeated until the occurrence of unacceptable toxicities, disease progression, or patient refusal. The primary end point was the overall survival. Results The response rates were 33.3%, 12.5%, and 0% after the first, second, and third line chemotherapy, respectively. The mean overall survival time was 994 days. The median survival time could not be calculated because 12 out of 19 patients were still alive when the study was concluded. Regarding hematological toxicity, the major adverse effect was leukopenia, which reached grades 3–4 in all lines of chemotherapy investigated. In addition, regarding non-hematological toxicities, the major adverse effect was anorexia, which reached grade 3–4 in the second line chemotherapy, and no deaths were attributable to the adverse effects of the drugs. Conclusion This sequential therapy was an effective treatment for advanced gastric cancer with acceptable toxic side-effects. We considered this sequential therapy to be effective because of the smooth switch to the next regimen

Annexin A4 is involved in proliferation, chemo-resistance and migration and invasion in ovarian clear cell adenocarcinoma cells.

Ovarian clear cell adenocarcinoma (CCC) is the second most common subtype of ovarian cancer after high-grade serous adenocarcinomas. CCC tends to develop resistance to the standard platinum-based chemotherapy, and has a poor prognosis when diagnosed in advanced stages. The ANXA4 gene, along with its product, a Ca(++)-binding annexin A4 (ANXA4) protein, has been identified as the CCC signature gene. We reported two subtypes of ANXA4 with different isoelectric points (IEPs) that are upregulated in CCC cell lines. Although several in vitro investigations have shown ANXA4 to be involved in cancer cell proliferation, chemoresistance, and migration, these studies were generally based on its overexpression in cells other than CCC. To elucidate the function of the ANXA4 in CCC cells, we established CCC cell lines whose ANXA4 expressions are stably knocked down. Two parental cells were used: OVTOKO contains almost exclusively an acidic subtype of ANXA4, and OVISE contains predominantly a basic subtype but also a detectable acidic subtype. ANXA4 knockdown (KO) resulted in significant growth retardation and greater sensitivity to carboplatin in OVTOKO cells. ANXA4-KO caused significant loss of migration and invasion capability in OVISE cells, but this effect was not seen in OVTOKO cells. We failed to find the cause of the different IEPs of ANXA4, but confirmed that the two subtypes are found in clinical CCC samples in ratios that vary by patient. Further investigation to clarify the mechanism that produces the subtypes is needed to clarify the function of ANXA4 in CCC, and might allow stratification and improved treatment strategies for patients with CCC

Involvement of phosphorylation, Ca²⁺ conjugation, and acetylation in production of two ANXA4 subtypes with different IEPs.

<p>(A) OVISE cells containing both acidic and basic ANXA4 subtypes were used to study effects of deacetylation, Ca²⁺ chelation, and dephosphorylation on subtype ratios. Each 2D-PAGE and western blot image for ANXA4 shows the treatment (above the panel), and the signal intensity (below each spot), as evaluated by an image analyzer ImageQuant LAS 4000 (GE Healthcare Life Science, UK). The left side spot indicates the acidic form and the right side spot indicates the basic form of ANXA4 respectively. (B) Acidic/basic subtype ratios were derived from signal intensities, and are shown in a bar of 100%.</p

ANXA4 expression visualized in surgically removed ovarian tumors using IHC.

<p>(A) Representative images for ANXA4 immunohistochemical (IHC) scores are shown. Each scale bar represents 200 µm. (B) IHC scores were significantly high in clear cell carcinoma compared with tumors with low malignant potential (LMP) and carcinomas (<i>P</i><0.05). ca, carcinoma; diff, differentiated.</p

Effect of ANXA4 knockdown on carboplatin and paclitaxel sensitivity.

<p>Sensitivity of OVTOKO cell to carboplatin (A) and paclitaxel (C); and of OVISE cells to carboplatin (B) and paclitaxel (D). Drug concentrations are indicated below each panel; cell viability under each concentration was evaluated by a WST-1 assay as described in the text. Experiments for clones and parental cells were performed in triplicate.</p

Effect of ANXA4 knockdown on cell migration and invasion and western blotting for membrane proteins RHAMM and LAMP2.

<p>OVISE cell migration (A) and invasion (B); and OVTOKO cell migration (C) and invasion (D). Experiments for clones and parental cells were performed in triplicate. (E) Expression of membrane proteins RHAMM and LAMP2 was demonstrated by western blotting in OVISE and OVTOKO cells. β-actin was used as a loading control in western blotting.</p

Establishment of ANXA4 knockdown clones and the effect on cell proliferation.

<p>(A) Western blotting for ANXA4, with β-actin as a loading control. Negative control clones, expressing non-<i>ANXA4</i> mRNA-targeting shRNA are designated as NC [clone number] with data for parental cells. (B, C) Proliferation was examined by a WST-1 assay as described in the text. Experiments for clones and parental cells were performed in triplicate.</p