Combination Of CDDP Research Articles

Hyperthermic potentiation of cisplatin by magnetic nanoparticle heaters is correlated with an increase in cell membrane fluidity

Magnetic fluid hyperthermia as a cancer treatment method is an attractive alternative to other forms of hyperthermia. It is based on the heat released by magnetic nanoparticles subjected to an alternating magnetic field. Recent studies have shown that magnetic fluid hyperthermia-treated cells respond significantly better to chemotherapeutic treatment compared with cells treated with hot water hyperthermia under the same temperature conditions. We hypothesized that this synergistic effect is due to an additional stress on the cellular membrane, independent of the thermal heat dose effect that is induced by nanoparticles exposed to an alternating magnetic field. This would result in an increase in Cis-diammine-dichloroplatinum (II) (cDDP, cisplatin) uptake via passive transport. To test this hypothesis, we exposed cDDP-treated cells to extracellular copper in order to hinder the human cell copper transporter (hCTR1)-mediated active transport of cDDP. This, in turn, can increase the passive transport of the drug through the cell membrane. Our results did not show statistically significant differences in surviving fractions for cells treated concomitantly with magnetic fluid hyperthermia and cDDP, in the presence or absence of copper. Nonetheless, significant copper-dependent variations in cell survival were observed for samples treated with combined cDDP and hot water hyperthermia. These results correlated with platinum uptake studies, which showed that cells treated with magnetic fluid hyperthermia had higher platinum uptake than cells treated with hot water hyperthermia. Changes in membrane fluidity were tested through fluorescence anisotropy measurements using trimethylamine-diphenylhexatriene. Additional uptake studies were conducted with acridine orange and measured by flow cytometry. These studies indicated that magnetic fluid hyperthermia significantly increases cell membrane fluidity relative to hot water hyperthermia and untreated cells, and hence this could be a factor contributing to the increase of cDDP uptake in magnetic fluid hyperthermia-treated cells. Overall, our data provide convincing evidence that cell membrane permeability induced by magnetic fluid hyperthermia is significantly greater than that induced by hot water hyperthermia under similar temperature conditions, and is at least one of the mechanisms responsible for potentiation of cDDP by magnetic fluid hyperthermia in Caco-2 cells.

Contribution of either YY1 or BclXL-induced inhibition by the NO-donor DETANONOate in the reversal of drug resistance, both in vitro and in vivo. YY1 and BclXL are overexpressed in prostate cancer

Nitric oxide (NO) donors have been shown to activate or inhibit constitutively-activated survival/anti-apoptotic pathways, such as NF-κB, in cancer cells. We report here that treatment of drug-resistant human prostate carcinoma cell lines with high levels (500–1000μM) of the NO-donor DETANONOate sensitized the resistant tumor cells to apoptosis by CDDP and the combination was synergistic. We hypothesized that DETANONOate inhibits previously identified NF-κB-regulated resistant factors such as Yin Yang 1 (YY1) and Bcl-2/BclXL. Lysates from tumor cells treated with DETANONOate showed inhibition of YY1 and BclXL expressions. Transfection with either YY1 or BclXL siRNA resulted in the inhibition of both YY1 and BclXL expressions and sensitized the cells to CDDP apoptosis. Mice bearing PC-3 tumor xenografts and treated with the combination of DETANONOate and CDDP resulted in significant inhibition of tumor growth; treatment with single agent alone did not have any effect on tumor growth. Analysis of patients TMA tissues with prostatic cancer revealed higher expression of both YY1 and BclXL as a function of tumor grades and their levels were directly correlated. Thus, both YY1 and BclXL are potential prognostic biomarkers. Overall, the above findings suggest that one mechanism of DETANONOate-induced sensitization of resistant tumor cells to CDDP correlated with the inhibition of NF-κB and its targets YY1 and BclXL. The examination of the combination of NO donors and cytotoxic therapy in the treatment of resistant prostate cancer may be warranted.

Starvation-induced activation of ATM/Chk2/p53 signaling sensitizes cancer cells to cisplatin

BackgroundOptimizing the safety and efficacy of standard chemotherapeutic agents such as cisplatin (CDDP) is of clinical relevance. Serum starvation in vitro and short-term food starvation in vivo both stress cells by the sudden depletion of paracrine growth stimulation.MethodsThe effects of serum starvation on CDDP toxicity were investigated in normal and cancer cells by assessing proliferation, cell cycle distribution and activation of DNA-damage response and of AMPK, and were compared to effects observed in cells grown in serum-containing medium. The effects of short-term food starvation on CDDP chemotherapy were assessed in xenografts-bearing mice and were compared to effects on tumor growth and/or regression determined in mice with no diet alteration.ResultsWe observed that serum starvation in vitro sensitizes cancer cells to CDDP while protecting normal cells. In detail, in normal cells, serum starvation resulted in a complete arrest of cellular proliferation, i.e. depletion of BrdU-incorporation during S-phase and accumulation of the cells in the G0/G1-phase of the cell cycle. Further analysis revealed that proliferation arrest in normal cells is due to p53/p21 activation, which is AMPK-dependent and ATM-independent. In cancer cells, serum starvation also decreased the fraction of S-phase cells but to a minor extent. In contrast to normal cells, serum starvation-induced p53 activation in cancer cells is both AMPK- and ATM-dependent. Combination of CDDP with serum starvation in vitro increased the activation of ATM/Chk2/p53 signaling pathway compared to either treatment alone resulting in an enhanced sensitization of cancer cells to CDDP. Finally, short-term food starvation dramatically increased the sensitivity of human tumor xenografts to cisplatin as indicated not only by a significant growth delay, but also by the induction of complete remission in 60% of the animals bearing mesothelioma xenografts, and in 40% of the animals with lung carcinoma xenografts.ConclusionIn normal cells, serum starvation in vitro induces a cell cycle arrest and protects from CDDP induced toxicity. In contrast, proliferation of cancer cells is only moderately reduced by serum starvation whereas CDDP toxicity is enhanced. The combination of CDDP treatment with short term food starvation improved outcome in vivo. Therefore, starvation has the potential to enhance the therapeutic index of cisplatin-based therapy.

Feasibility of postoperative adjuvant chemotherapy of cisplatin plus vinorelbine for completely resected non-small-cell lung cancer: A retrospective study in Japan

The efficacy of postoperative adjuvant cisplatin (CDDP)-based chemotherapy, such as the combination of CDDP and vinorelbine (VNR), has been established for surgically resected non-small-cell lung cancer (NSCLC). However, the optimal treatment schedule and dosage for CDDP and VNR are unknown. We evaluated patient compliance with and the safety of adjuvant chemotherapy of CDDP at 80 mg/m(2) administered on day 1 plus VNR at 25 mg/m(2) administered on days 1 and 8, every 3 weeks. Medical records of 100 surgically resected NSCLC patients, treated with a combination of CDDP and VNR at the Shizuoka Cancer Center between February 2006 and October 2011, were retrospectively reviewed. Eighty-three (83%) patients completed the planned 4 cycles of CDDP plus VNR and 59 (59%) received the planned doses. Sixty-eight percent of the patients experienced a decreased neutrophil count (grade 3/4 toxicity); 1%, a decreased platelet count; and 4%, febrile neutropenia. No treatment-related deaths were noted in this study. Univariate analysis of the factors influencing patient compliance with this adjuvant chemotherapy showed that neither patient characteristics nor surgical procedure was significantly associated. Our results indicated that adjuvant chemotherapy with CDDP at 80 mg/m(2) administered on day 1 plus VNR at 25 mg/m(2) administered on days 1 and 8, every 3 weeks, was feasible for surgically resected NSCLC cases.

Differential Effects of Lovastatin on Cisplatin Responses in Normal Human Mesothelial Cells versus Cancer Cells: Implication for Therapy

The cancer killing efficacy of standard chemotherapeutic agents such as cisplatin (CDDP) is limited by their side effects to normal tissues. Therefore, research efforts optimizing the safety and efficacy of those agents are clinically relevant. We did screen for agents that specifically protect normal human mesothelial cells against CDDP without reducing the cancer cell killing efficacy. Lovastatin was identified from the screen. Lovastatin at a pharmacologically relevant concentration strongly arrested the proliferation of normal cells, whereas cancer cells were less affected. CDDP-induced DNA damage response was not activated and normal cells showed enhanced tolerance to CDDP when normal cells were treated with the combination of CDDP and lovastatin. We demonstrate that interfering with protein geranylgeranylation is involved in the lovastatin-mediated CDDP protective effect in normal cells. In contrast to normal cells, in cancer cells lovastatin did not change the CDDP-induced response, and cancer cells were not protected by lovastatin. Furthermore, lovastatin at the pharmacological relevant concentration per se induced DNA damage, oxidative stress and autophagy in cancer cells but not in normal mesothelial cells. Therefore, our data suggest that lovastatin has a potential to improve the therapeutic index of cisplatin-based therapy.

158. Validity of the combination treatment of the surgery and chemotherapy in advanced gastric cancer

Background: While surgery and appropriate adjuvant chemotherapy have resulted in superior stage-by-stage survival in Japan, the prognosis of stage IV advanced gastric cancer remains unsatisfied. Phase III trials conducted in Japan for unresectable or recurrent gastric cancer have emphasized the utilities of the S1 monotherapy and a combination of S1 and CDDP, other combination therapies. Therefore, the purpose of this study reveals how combination of an operation and antineoplastic drug medical treatment should be performed for the purpose of long-term survival in a bad prognosis's stage advance cancer case.

P-0039 Irinotecan as a Second/Third Line Chemotherapy in Gastric Cancer Patients who Failed S-1-Based Treatment

IntroductionUnresectable/metastatic gastric cancer patients have been treated with S-1 or S-1/CDDP combination in Japan. More recently, evidence has evolved that a second line treatment with irinotecan is superior in terms of survival time to the best supportive care among patients who failed a first line treatment. Efficacy of the second line after failing S-1 remains unknown. UGT1A1 genotyping is known to predict some of the adverse reactions. MethodsA phase II trial was conducted to evaluate the efficacy and safety of intravenous administration of irinotecan at 100 mg/m2 on days 1, 7, 14 every 4 weeks. Patients who failed treatment with a S-1 containing regimen were eligible. Another line of treatment was allowed before entering onto this study. All patients were to undergo genotyping of UGT1A1*6, *27 and *28, and only those with wild type gene and those heterozygous for either *6 or *28 were rendered eligible. The primary endpoint was response rate, assessed in accordance with the RECIST v1.1. A Simon optimal 2-stage design was adopted assuming unacceptable and acceptable response rates of 4% and 20% respectively, and 5% type I error and 20% type II error rates. Required sample size was 27 patients. Treatment-related toxicities were assessed according to the CTC-AE v3.0. ResultsForty-one patients underwent genotyping and 5 patients were excluded because of the inadequate genotype. Six other patients failed to receive irinotecan due to rapid disease progression prior to initiation of the treatment etc. Of the 30 patients who received treatment, 21 received irinotecan as the second line and 9 as the third line treatment. The median number of the 4-week treatment administered was 3 cycles. One CR, 3 PRs, 14 SDs and 6 PDs were recorded for the overall response rate of 13% and disease control rate of 60%, including 6 patients who failed to undergo formal evaluation by the imaging studies. Six of 7 patients who was heterozygous for UGT1A1*6 had either significant tumor shrinkage or stable disease. Grade III/IV toxicities included neutropenia (30%), anorexia (23%), leucopenia (17%), diarrhea (17%), anemia (13%), fatigue (13%), vomiting (10%) and thrombocytopenia (3.3%). Median survival time of all patients was 284 days, 342 days for the second line and 218 days for the third line treatment. ConclusionIrinotecan as a single agent was modestly active in the second/third line setting for advanced gastric cancer after failing S-1, with the response rate of 13% and disease control rate of 60%.

Phase I trial of everolimus, gemcitabine and cisplatin for patients with solid tumors refractory to standard therapy.

e13052 Background: The combination of GEM and CDDP has shown activity in a variety of cancers, including biliary tract/gallbladder. Preclinical testing shows a potential benefit to the addition of EV. Methods: Using a standard 3+3 design, the MTDs of GEM + EV (Cohort I) and GEM + CDDP + EV (Cohort II) were determined as shown in the table. The MTD was defined as the dose level below the lowest dose that induces a DLT in at least one-third of pts. At the MTD of Cohort II, 10 patients were enrolled with biliary tract/gallbladder cancer (Cohort III). A weekly CBC was obtained. Assessments occurred every 3 weeks and imaging for response at every other cycle. Results: In Cohort I (N=12), no DLT occurred at dose level 0, and in dose level 1, grade 3 thrombocytopenia was found in 2 of 6 pts. The MTD for Cohort I was determined to be dose level 0. Responses were seen in 3 pts: 2 CRs (primary peritoneal, pancreatic) and 1 PR (breast). In Cohort II (N=15) DLTs at dose level 0 were neutropenia and thrombocytopenia in 2 of 3 pts and at dose level -1, thrombocytopenia in 2 of 6 pts. At dose level -2, 1 of 6 pts experienced a DLT (grade 3 thrombocytopenia), establishing this dose level as the MTD. Responses were seen in 2 pts, both PRs (ampullary, pheochromocytoma). All 10 pts have been enrolled in Cohort III with 2 DLTs (neutropenia and thrombocytopenia). Stable disease seen in 5 of 7 evaluable pts. Conclusions: Gem + EV was well tolerated at dose level 0, though dose escalation was limited by thrombocytopenia. GEM + CDDP + EV had DLTs of neutropenia and thrombocytopenia leading to the MTD of dose level of -2. Cohort III is fully accrued and the 2 DLTs are hematologic. [Table: see text]

A phase II study of cisplatin (CDDP) and oral vinorelbine (NVBO) concomitantly with radiotherapy (RT) in locally advanced (LA) non-small cell lung cancer (NSCLC) treatment: Interim analysis.

e17545 Background: CDDP+NVBO as induction and concomitant regimen with RT has shown good efficacy outcomes and safety profile (Krzakowski, J Thor Oncol. 2008). The objective of this study was to evaluate the effectiveness and toxicities of the combination of CDDP and NVBO given at full doses concomitantly with RT in LA-NSCLC. Methods: Between February 2010 and May 2011, 26 chemo-naive patients (p) with histologically confirmed stage IIIA/IIIB unresectable LA NSCLC were treated. Treatment consisted of 4 cycles (cy) of NVBO 60 mg/m2 on days 1 and 8 and CDDP 80 mg/m2 every 3 weeks plus RT 66 Gy starting on day 1, cy 2. A standard Fleming two stage design is being used. The sample size is calculated with a power of 90% and a two-sided type 1 error allowance of 5%. Stage 1 included 25 pts, with subsequent expansion to a total of 45. Results: Patient’s characteristics were: Median age, 59 years (range 48-71); males, 92%; smokers, 60%; adenocarcinoma, 28% / squamous, 64%; stage IIIA, 54% / IIIB, 46%. 21 p completed the 4 cy treatment as per protocol. We analyzed 92 cy. Hematological toxicities (% cy): grade (g) 3/4 neutropenia, 42.3%; g3 anemia, 3.3%; g3 thrombocytopenia, 2.2%; febrile neutropenia, 1 p. Non-hematological toxicities (% cy): g3 oesophagitis, 6.5%; g4 diarrhea, 1.1%. 21 p were evaluable for response. 1 CR (4.8%), 17 PR (81%) and 1 SD (4.5%) were reported. ORR 85.8%. Survival analysis has not been performed due to short follow-up. Conclusions: The findings of this interim analysis confirm the previously reported outcomes of efficacy and safety with NVBO plus CDDP when administered concurrently with RT in stage IIIA/IIIB p. The trial has met the criteria for continuation into stage 2.

Abstract 4337: Effect of sorafenib in combination with cytostatic agents on hepatoblastoma in vitro and in vivo

Abstract Background: Hepatoblastoma (HB) represents the most common malignant liver tumour in childhood. For patients with high risk or recurrent HB treatment results remain still poor due to a developing multidrug resistance during chemotherapy. Therefore new therapy strategies have to be evaluated. The primary objective was to investigate the combined therapy of cytotoxic agents with sorafenib. Sorafenib has been shown to reduce tumour progression and angiogenesis. In order to investigate a treatment close to a clinical setting the effect of the combined administration of cytotoxic agents with sorafenib on HB cell lines and xenotransplanted HB tumours was determined. Methods: Cell viability of two HB cell lines (HUH6 and HepT1) was evaluated in using MTT assays after treatment with sorafenib and different cytostatic agents (cisplatin (CDDP), doxorubicin, irinotecan, topotecan). ERK signalling was investigated by Western blot analysis. NMRI mice (nu/nu) bearing subcutaneous HUH6 derived tumours were treated with Sorafenib orally (30 mg/kg, on days 4-13 and 17-24) alone or in combination with CDDP i.p. (3 mg/kg, on days 1-3 and 14-16). Tumor progression and viability were monitored by tumour volume and AFP levels. Apoptosis was assessed by TUNEL assay. Angiogenesis was determined by CD31 staining and mean vascular density (MVD) was calculated. Results: In vitro the combination of sorafenib with CDDP showed a remarkable decrease in cell viability compared to the other agents tested. However with increasing concentrations of CDDP this additive effect disappeared. CDDP alone showed an enhanced phosphorylation of ERK1/2, which is a common pathway for apoptosis induction through NOXA. CDDP in combination with sorafenib led to an overall reduced phosphorylation level of ERK1/2, which also resulted in a low HB cell viability. Since CDDP and sorafenib have an opposing effect on phosphorylation of ERK1/2, we avoid the concomitant administration of both drugs in vivo. In HB xenografts both sorafenib alone or in alternating combination with CDDP reduced tumour growth compared to the control untreated group (p < 0.05). The combination treatment showed a higher and earlier reduction of tumor volume than sorafenib alone. In addition AFP levels did not significantly change in the treatment groups. Apoptotic areas as determined by TUNEL analysis increased significantly after administration of sorafenib or sorafenib and CDDP (p < 0.08). Furthermore CD31 staining showed a significant inhibition of angiogenesis as revealed by determination of the MVD in both treatment groups (p < 0.02). Discussion: Taken together the combination therapy of Sorafenib and CDDP led to an enhanced decrease in cell viability as well as in tumour progression and angiogenesis than sorafenib alone. Therefore a combined therapy might be a promising option in the treatment of high risk or recurrent hepatoblastoma. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4337. doi:1538-7445.AM2012-4337

Effect of a combination of aurora kinase A inhibitor MLN-8237 and cisplatin on gastrointestinal tumors.

58 Background: Gastric cancers respond poorly to current chemotherapeutic protocols indicating active intrinsic mechanisms against drug induced cell death. We have previously reported that AURKA overexpression induces pro-survival (AKT and β-catenin) and inhibits apoptotic (p53 and p73) pathways. Methods: We evaluated the effect of AURKA specific inhibitor (MLN-8237) alone and/or in combination with cisplatin (CDDP) in gastrointestinal (GI) cancer cells. Following treatment with MLN-8237 and/or CDDP, cell survival, cell viability, western blot and qRT-PCR analyses were done on GI cancer cell lines. Additionally, tumor xenografted mice were treated with MLN (30mg/kg) and/or CDDP (2mg/kg) for 21 days and tumor volume was measured every alternate day. Results: The ATP-Glo cell viability and cell survival assay data indicated that in comparison to single agent treatments alone; the combination treatment with MLN-8237 (0.5µM) and CDDP (2.5µM) significantly increased inhibition of cellular viability and survival (p<0.05) in AGS, KATO-III, MKN28, FLO-1, OE-19 and OE33 cell lines. Similarly, the western blot data exhibited higher levels of p73, Puma, Noxa, cleaved caspase 3 and cleaved PARP with MLN (0.5µM) and CDDP (2.5µM) combination treatment, as compared to single agent treatments in AGS, FLO-1 and OE33 cell lines. The tumor growth results for OE33 mouse xenograft study demonstrate synergistic antitumor effect with MLN-8237 and CDDP combination in the following order of effectiveness: Control > CDDP > MLN-8237 > and MLN-8237/CDDP (p<0.01). Moreover, the qRT-PCR data from tumor xenograft samples treated with MLN-8237/CDDP combination showed significantly higher mRNA levels of p73 downstream transcriptional target genes (P21, PUMA, NOXA and BAX) (p<0.05). Conclusions: Our in vitro and in vivo data indicate that inhibition of AURKA with MLN-8237 in combination with CDDP enhances the antitumor activity of MLN-8237 against GI cancer cells. Therefore, our study demonstrates that MLN-8237 is an effective anti-tumor agent which can be potentially combined with CDDP for better therapeutic outcome in GI cancer patients.

Cytotoxic Activity of Anti-CD20-hIFN-α on Rituximab-Resistant B-NHL Clones and Synergy with Chemotherapy,

Abstract 3499Rituximab (chimeric anti-CD20 mAb) (Rtx) has been successfully used in the treatment of patients with B-cell non-Hodgkin’s lymphomas (B-NHLs). The combination treatment with chemotherapy results in achieving high response rates and prolongation of survival. However, a subset of patients does not initially respond to treatment and many responding patients relapse and no longer respond to further treatments. Currently, there are no alternative therapies for resistant patients. The mechanism of resistance in vivo is not clear. However, we have explored a potential mechanism by developing in vitro several clones of Rtx-resistant (RR) variants for several B-NHL cell lines and characterized their properties. Briefly, unlike the parental wild-type, the RR clones express CD20 but no longer respond to treatment with Rtx or combination with cytotoxic drugs. Further, these clones overexpress the activity of several survival/anti-apoptotic pathways [1]. It is not known whether chemical modification of Rtx might be necessary to exert its activity and signaling on the RR clones. Hence, a recent report demonstrated that a fusion protein consisting of Rtx and human IFN-α (anti-CD20-hIFN-α) exhibited superior activity over Rtx, IFN-α, or combination of Rtx and IFN-α, and exhibited significant anti-proliferative and apoptotic effects in vitro with several B-NHL cell lines and in vivo an anti-tumor xenograft response [2]. These findings prompted us to investigate the effect of anti-CD20-hIFN-α on the RR clones. We hypothesized that anti-CD20-hIFN-α may exert an anti-proliferative and apoptotic effects on the RR clones and may also synergize when used in combination with chemotherapy. In this study, we used the B-NHL line Ramos (Burkitt) and 2F7 (AIDS-related) and their respective Ramos RR1 and 2F7 RR1 clones as models. We examined the effects of anti-CD20-hIFN-α and Rtx on the wild-type and RR clones following treatment with IgG isotype control, Rtx, anti-CD20-hIFN-α, CDDP (10 mg/ml) and Treanda® (Bendamustine) (5 mg/ml), as well as combinations. Treatment of 2F7 with single agents alone had no cytotoxic effect; however, treatment with the combination of Rtx and CDDP or Treanda® or anti-CD20-hIFN-α plus CDDP or Treanda® resulted in significant cytotoxicity. Treatment of Ramos resulted in similar findings observed with 2F7, however, the anti-CD20-hIFN-α alone was significantly cytotoxic to Ramos cells. Importantly, whereas treatment of 2F7 RR1 or Ramos RR1 with Rtx or Rtx plus CDDP or Treanda® had no cytotoxic effects (as expected), the treatment with the anti-CD20-hIFN-α alone had significant cytotoxicity and synergy was observed when used in combination with CDDP or Treanda®. In all of the above experiments, the level of cytotoxicity was a function of the antibody concentration used (range 10–30 μg/ml). The mechanism by which anti-CD20-hIFN-α signals the RR clones for cytotoxicity and sensitization was examined. Preliminary findings show that treatment of the RR clones with anti-CD20-hIFN-α inhibits the activity of p38MAPK survival pathway and also inhibits the anti-apoptotic gene products, Bcl-2/BclXL and upregulates the pro-apoptotic expression of Bax. These findings established, for the first time, that modification of Rtx by fusion with IFN-α was cytotoxic on the RR clones and synergized with chemotherapy. The findings also show, unlike Rtx that, anti-CD20-hIFN-α signals the RR cells and inhibits survival/antiapoptotic pathways leading to direct cytotoxicity and chemo-sensitization. The molecular signaling mediated by anti-CD20-hIFN-α on the cell membrane of RR cells leading to inhibition of survival pathways will be presented. Disclosures:No relevant conflicts of interest to declare.

The synergistic therapeutic effect of cisplatin with Human papillomavirus E6/E7 short interfering RNA on cervical cancer cell lines in vitro and in vivo

Human papillomavirus (HPV) types 16 and 18 are the major etiologic factors in the development of cervical epithelial neoplasia. Our study was designed to validate antiviral short interfering RNA (siRNA) targeting the E6 and E7 oncogenes as a potential chemosensitizer of cisplatin (cis-diaminedichloroplatinum II; CDDP) in cervical carcinoma. Specifically, the therapeutic efficacy of combination of CDDP and E6/E7-specific siRNA was assessed in an in vivo cervical cancer xenograft models. The combination of CDDP and E6/E7-specific siRNA had greater efficacy than the combination of CDDP and E6-specific siRNA especially in terms of inducing cellular senescence. Through in vitro and in vivo experiments, the mechanism of synergy between these two treatments was revealed, demonstrating that the combination of E6/E7-specific siRNA and CDDP therapy was significantly superior to either modality alone. In vitro, long-term exposure of HeLa cells to the combination of CDDP and E6/E7-specific siRNA induced apoptosis and cellular senescence. In vivo, E6/E7-specific siRNA potentiated the antitumor efficacy of CDDP via induction of apoptosis, senescence and antiangiogenesis. Our results suggest that E6/E7-specific siRNA may be an effective sensitizer of CDDP chemotherapy in cervical cancer.

Cell-growth and migration inhibition of human mesothelioma cells induced by 3-O-Methylfunicone from Penicillium pinophilum and cisplatin

Malignant pleural mesothelioma is a fatal malignancy linked to asbestos exposure. The main challenge for mesothelioma treatment is to go beyond the drug resistance, in particular against cisplatin (CDDP), one of the most used chemotherapeutic drug. 3-O-methylfunicone (OMF) is a metabolite produced by the fungus Penicillium pinophilum; its antiproliferative properties have been previously studied in vitro. Particularly, OMF is able to inhibit mesothelioma cell motility. To improve the effects of CDDP by-passing the resistance of mesothelioma cells to this drug, in the present study we investigated the combined treatment of OMF with CDDP respectively in an established mesothelioma cell line (NCI) and primary mesothelioma cells (Mest). As compared to the effect of single treatments, the combination of OMF and CDDP resulted in a stronger inhibition of NCI and Mest cell proliferation. OMF combination with CDDP was also able to affect the migratory ability of NCI and Mest cells by down-regulating αv and β5 expression and reducing metalloproteinase 2 (MMP-2) production. In addition, this association was effective in modulating VEGF gene expression. This finding highlights the possibility to use OMF and CDDP together to regulate angiogenesis and tumour progression in mesothelioma.

Cisplatin (CDDP) plus oral vinorelbine (NVBO) as first-line treatment for advanced non-small cell lung cancer (NSCLC): Prospective analysis to improve the patient’s convenience on day 8 NVBO administration.

e18060 Background: Severe neutropenia observed during chemotherapy (CT) is a clinical finding leading to treatment modification and, sometimes, life-threatening events. The results of a previous study with 180 p treated with IV vinorelbine plus CDDP as first-line treatment for advanced NSCLC could lead to consider not performing a blood count prior to day 8 vinorelbine administration for patients aged 70 years (y) or less who presented a good haematological tolerability profile during previous cycles (cy) in order to improve treatment convenience. The aim of this study is to prospectively validate these results with the combination of NVBO plus CDDP. Methods: Between October 2007 and March 2009, 56 chemo-naïve p with histologically confirmed stage IIIB/IV NSCLC were included. Treatment consisted of CDDP 75 mg/m2 day 1 plus NVBO 80 mg/m2 days 1 and 8, with a prior test of myelosensitivity at 60 mg/m2 for the first cycle, every 21 days. Patient’s characteristics were: Median age, 61 years (range 32-75); males, 85.7%; smokers, 35.7%; all PS 0-1; adenocarcinoma, 37.5% / squamous, 35.7%; stage IIIB, 17.9% / IV, 75%. Results: Patients received a total of 251 cycles. For the purpose of this report 181 were analyzed (cycle 1 from each p was excluded). NVBO administration on day 8 was cancelled in 4 cycles due to haematological events (2.2% [95% CI: 0.6%, 5.56%]). The reported events were: 3 grade 2-4 neutropenia and 1 grade-2 thrombocytopenia. Among 54 p evaluable for response, 1 p achieved complete response and 21 p partial response (overall response rate, 40.7% [95% CI: 27.6-55]). Toxicity was mild with neutropenia WHO grade 3-4 observed in 2.8% cy (12.5% p), leukopenia grade 3-4 in 1.6% cy (7.2% p); 2 cases of febrile neutropenia were reported. Grade-3 fatigue was observed in 9 cy (8 p), grade-3 nausea in 4 cy (4 p) and grade 3-4 vomiting in 5 cy (5 p). Conclusions: The findings of this study could lead to consider not performing a blood count prior to day 8 NVBO administration for patients aged 70 y or less who presented a good haematological tolerability profile during previous cycles in first-line treatment for advanced NSCLC.

Abstract 1689: The combination treatments using siRNA and CDDP are effective therapeutic options for cancers

Abstract Background: RNA interference technology has been developed as potential therapeutic options for several diseases, including cancer. Short interfering RNA (siRNA) is the best candidate for cancer therapy because of its specificity and efficiency in gene-specific silencing. We reported that the efficiency of siRNA as novel therapeutic agents for several cancers, such as cholangiocarcinoma, breast cancer and colorectal cancer. Methods: We examined the effect of a combination treatment using Nek2 (NIMA related kinase 2) siRNA with anti cancer agent, CDDP (Cisplatin) on xenograft colorectal cancer mouse model. We next examined the effect of a combination treatment using Tlk1 (Tousled like kinase 1) siRNA with CDDP on MTT assay and TUNEL assay on cholangiocarcinoma cell lines. Results and Discussion: The combination treatment using Nek2 siRNA and CDDP suppressed the tumor growth in colorectal cancer model, more efficiently than negative control siRNA and CDDP. It was more effective than the treatment with Nek2 siRNA alone. The combined administration of Tlk1 siRNA and CDDP strongly induced apoptosis in comparison with negative control siRNA and CDDP on cholangiocarcinoma cell lines. Tlk1 siRNA enhanced DNA damage induced by CDDP treatment. The combination of siRNA and CDDP was effective therapeutic options in colorectal cancer and cholangiocarcinoma. Conclusions: Our data provides a strong rationale for the further investigation concerning the combination of both siRNA and anti cancer drug as a new cancer strategy, holding the potential to provide regression of multiple human malignancies. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1689. doi:10.1158/1538-7445.AM2011-1689

Abstract 1755: Sensitization of radiotherapy and chemotherapy by adjunction of R1507 (anti-IGF-1R monoclonal antibody) in small cell lung cancer cell lines: Opportunity of translating the triple combination R1507-Cisplatin-radiotherapy

Abstract Introduction: Insulin like growth factor receptor-1 (IGF-1R) inhibition could be a pertinent therapeutic approach in small cell lung cancer (SCLC), given the recognized activation of an IGF-1R autocrine loop. Materials and methods: We evaluated the importance of IGF-1R axis in SCLC by assessing IGF-1R expression and Akt activation in 83 human SCLC tissue specimens. In parallel, we evaluated in vitro the efficacy of R1507, an IgG1 fully human monoclonal antibody directed to IGF-1R. R1507 effects on IGF-1R and downstream pathways were assessed using three SCLC cell lines: H69, H146 and H526. The cytotoxicity of R1507, Cisplatin and ionizing radiation (IR) was evaluated by WST-1 cell proliferation assays and clonogenic survival assays. In vivo, the efficacy of R1507, Cisplatin and IR was assessed on H526 and H146 xenografts in nude mice. Results: IGF1R was overexpressed and Akt was activated in 38 (46%) and 31 (37%) of 83 SCLC tumors, respectively. In vitro, R1507 down-regulates IGF-1R receptor and disrupts downstream signaling through Akt and MAPK pathways in a dose dependent manner in selected small cell lung cancer cell lines. R1507 inhibits cell proliferation and colony forming capacity in H526 and H146 cells but not in H69 cells. The inhibition of PI3K-Akt pathway correlated with treatment response. The combination of R1507 (200 nM) and CDDP (3 μM) exhibited a synergistic inhibitory effect on the colony forming capacity of H146 cells and an additive inhibitory effect on H526 cells. R1507 showed synergistic effects with IR (2 Gy) in H146 cells and an additive inhibitory effect in H526 cells. R1507 potentiates the IR effects and induces a prolonged increase of the proportion of sub G1 cells in H526 but not in H146 cell line. The triple combination R1507-Cisplatin-IR potentiates the antitumoral effect of Cisplatin-IR (current standard treatment) in H146 and in H526 cell lines. In vivo, R1507 administered in monotherapy leads to a non-significant delay of tumor growth in H526 xenografts but sensitizes to both Cisplatin and IR. The triple combination R1507-Cisplatin-IR achieves a long lasting delay in tumor growth as compared with the current standard treatment Cisplatin-IR. Conclusion: R1507 has a single agent activity and remarkable chemo- and radiosensitizing effect in defined SCLC cell lines in vitro. Efficacy is dose-dependent and related to the capacity to inhibit PI3K-Akt signaling pathway. In vivo, it sensitizes to both IR and Cisplatin effects. The triple combination R1507-Cisplatin-IR exhibits a long lasting tumor growth delay as compared with the current standard treatment Cisplatin-IR, the potential of this combination should be evaluated in further early clinical trials. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1755. doi:10.1158/1538-7445.AM2011-1755

A Case of Advanced Gastric Cancer with Liver Metastases Resected Successfully after S-1+CDDP Combination Chemotherapy

症例は63歳男性. 上腹部不快感を主訴に近医を受診した. 上部消化管内視鏡検査で胃前庭部小弯に, 2型の腫瘍を認め, 生検の結果, 中～低分化管状腺癌と診断され, 石井病院外科に紹介された. CT検査で肝左葉に大きさ約5 cmの低吸収領域を認め, 胃癌の肝転移と診断した. 平成22年4月からS-1＋CDDP療法を開始した. 7コース終了後の内視鏡では腫瘍は明らかに縮小傾向で, 瘢痕組織様であった. またMRIで肝転移の著明な縮小がみられ, 腫瘍は外側区域に限局した. 平成23年3月群馬大学医学部附属病院で幽門側胃切除D2郭清, Billroth I法再建, 肝外側区切除を施行した. 病理学的にはtub2, pT2 (MP), pN0, INFβ, ly0, v2, PM (－), DM (－), H1, pStage IVであったがR0切除ができた. 経過は良好で, 第12病日に退院した. その後石井病院外来で経過観察中であるが, 術後5ヶ月の現在明らかな再発はない.

Antitumor and immunoregulatory effects of astragalus on nasopharyngeal carcinoma In Vivo and In Vitro

This study was carried out to evaluate the effects of Astragalus on human nasopharyngeal carcinoma (NPC) viability and apoptosis and to investigate the mechanism of Astragalus in a NPC cell line (CNE2). Cell viability was measured using the MTT assay. CNE2 cells treated with Astragalus were stained with acridine orange/ethidium bromide and subjected to fluorescence microscopy. Bcl-2, Bax, caspase-3 and -8 were measured by western blotting. Rat NPC cells were used to establish a NPC model. Tumor weight, immune organ index and T lymphocyte subsets were employed to detect the immunoregulatory and antitumor effects of Astragalus after administration. Astragalus was effective in inducing apoptosis in CNE2 cells. Morphological changes associated with cell injury were found. Western analysis showed caspase-3, -8, and Bax protein levels were increased after Astragalus treatment, while the bcl-2 protein level was decreased. Astragalus increased the percentage of CD3(+) , CD4(+) T-lymphocytes, and the ratio of CD4(+) /CD8(+) . Astragalus also restored the immunological effects of DDP-induced immunosuppression. These findings suggest that the immunomodulatory and anticancer effects of DDP + Astragalus were better than those of DDP alone, and Astragalus could inhibit immunosuppression induced by DDP. The combination of CDDP + Astragalus could be developed as an effective chemotherapeutic regimen in the treatment of nasopharyngeal carcinoma.

Restoration of the immunogenicity of cisplatin-induced cancer cell death by endoplasmic reticulum stress

In contrast to other cytotoxic agents including anthracyclins and oxaliplatin (OXP), cisplatin (CDDP) fails to induce immunogenic tumor cell death that would allow to stimulate an anticancer immune response and hence to amplify its therapeutic efficacy. This failure to induce immunogenic cell death can be attributed to CDDP's incapacity to elicit the translocation of calreticulin (CRT) from the lumen of the endoplasmic reticulum (ER) to the cell surface. Here, we show that, in contrast to OXP, CDDP is unable to activate the protein kinase-like ER kinase (PERK)-dependent phosphorylation of the eukaryotic translation initiation factor 2α (eIF2α). Accordingly, CDDP also failed to stimulate the formation of stress granules and macroautophagy, two processes that only occur after eIF2α phosphorylation. Using a screening method that monitors the voyage of CRT from the ER lumen to the cell surface, we identified thapsigargin (THAPS), an inhibitor of the sarco/ER Ca(2+)-ATPase as a molecule that on its own does not stimulate CRT exposure, yet endows CDDP with the capacity to do so. The combination of ER stress inducers (such as THAPS or tunicamycin) and CDDP effectively induced the translocation of CRT to the plasma membrane, as well as immunogenic cell death, although ER stress or CDDP alone was insufficient to induce CRT exposure and immunogenic cell death. Altogether, our results underscore the contribution of the ER stress response to the immunogenicity of cell death.