Molecular Mechanism Of Anti-tumor Effect Research Articles

Induction of different cellular arrest and molecular responses in low EGFR expressing A549 and high EGFR expressing A431 tumor cells treated with various doses of 177Lu-Nimotuzumab

Introduction Radioimmunotherapy (RIT) is a major anti-cancer therapy in cancer management multimodalities. 177Lu-Nimotuzumab has been in the use for radioimmunotherapy of EGFR expressing tumors. This study focuses on understanding the differential cellular and molecular mechanisms of anti-tumor effects of 177Lu-Nimotuzumab on low EGFR expressing A549 and high EGFR expressing A431 tumor cells. Materials and methods Nimotuzumab labeled with 177Lu was characterized by SE-HPLC. Specificity of 177Lu-Nimotuzumab to EGFR expressed on A549 and A431 cells was confirmed by competitive assay using increasing amounts of unlabeled Nimotuzumab. Cellular responses of A549 (low EGFR) and A431 (high EGFR) in response to different doses of 177Lu-Nimotuzumab were determined by Viable count assay for cellular viability, cell-cycle analysis by DNA staining, apoptotic assay for cell death, and CFSE dilution assay for cellular proliferation capacity. The number of DNA DSBs formed was determined using γ-H2AX assay with PI staining. Transcription of genes involved in DNA damage response and repair (DRR) pathways was monitored by RT-qPCR. Results 177Lu-Nimotuzumab characterized by SE-HPLC exhibited a radiochemical purity of 99.1 ± 0.6%. Cell binding competition studies with 177Lu-Nimotuzumab showed specific binding of 34.3 ± 1.7% with A431 cells and 18.4 ± 1.9% with A549 cells which decreased when co-incubated with unlabeled Nimotuzumab. Cytotoxicity and DNA damage (DNA DSBs) increased with an increase in the dose of 177Lu-Nimotuzumab. A549 displayed G2/M arrest while A431 showed G1 arrest. Apoptotic death was determined to be one of the modes of death of arrested A549 and A431 cells which increases with the increase in the dose of 177Lu-Nimotuzumab. Loss of proliferation capacity was higher in A431 showed by CFSE staining at different doses of 177Lu-Nimotuzumab. Transcription profile of most DRR genes in A431 and A549 showed a decrease in the transcription at 4 h followed by recovery at 16 h post-treatment. The degree of transcription of most DRR genes was similar, irrespective of 177Lu-Nimotuzumab dose. Conclusion 177Lu-Nimotuzumab induces different cellular arrest and molecular responses in low EGFR expressing A549 and high EGFR expressing A431 tumor cells. This study would enable the development of integrative novel treatment strategies for radioimmunotherapy in low and high EGFR expressing tumors by 177Lu-Nimotuzumab with therapeutic gains.

Establishment and characterization of NCC-ASPS1-C1: a novel patient-derived cell line of alveolar soft-part sarcoma.

Alveolar soft-part sarcoma is a mesenchymal malignancy characterized by the rearrangement of ASPSCR1 and TFE3 and a histologically distinctive pseudoalveolar pattern. Although alveolar soft-part sarcoma takes an indolent course, its long-term prognosis is poor because of late distant metastases. Currently, curative treatments have not been found for alveolar soft-part sarcoma, and hence, a novel therapeutic strategy has long been required. Patient-derived cell lines comprise an important tool for basic and preclinical research. However, few cell lines from alveolar soft-part sarcoma have been reported in the literature because it is an extremely rare malignancy, accounting for less than 1% of all soft-tissue sarcomas. This study aimed to establish a novel alveolar soft-part sarcoma cell line. Using surgically-resected tumor tissue of alveolar soft-part sarcoma, we successfully established a cell line and named it NCC-ASPS1-C1. The NCC-ASPS1-C1 cells harbored an ASPSCR1-TFE3 fusion gene and exhibited slow growth, and spheroid formation. On the other hand, NCC-ASPS1-C1 did not show the capability of invasion. We screened the antiproliferative effects of 195 anticancer agents, including Food and Drug Administration-approved anticancer drugs. We found that the MET inhibitor tivantinib and multi-kinase inhibitor orantinib inhibited the proliferation of NCC-ASPS1-C1 cells. The clinical utility and molecular mechanisms of antitumor effects of these drugs are worth investigating in the further studies, and NCC-ASPS1-C1 cells will be a useful tool for the in vitro study of alveolar soft-part sarcoma.

Vitamin D and estrogen-dependent tumors

Vitamin D is characterized by undeniable antitumor properties that are important for both prevention and treatment of tumor diseases. The results of a systematic analysis of the molecular mechanisms of antitumor effects of vitamin D (inhibition of the Wnt/b-catenin cascade, the transcription factor WSTF) and the results of clinical studies of patients with estrogen-dependent tumors are presented, indicating the promise of combined use of estrogen and vitamin D.

Molecular mechanism of anti-tumor effect by heperthermia

Molecular mechanism of anti-tumor effect by heperthermia

Effects of JS-K, a novel anti-cancer nitric oxide prodrug, on gene expression in human hepatoma Hep3B cells.

JS-K is a novel anticancer nitric oxide (NO) prodrug effective against a variety of cancer cells, including the inhibition of AM-1 hepatoma cell growth in rats. To further evaluate anticancer effects of JS-K, human hepatoma Hep3B cells were treated with JS-K and the compound control JS-43-126 at various concentrations (0-100μM) for 24h, and cytotoxicity was determined by the MTS assay. The compound control JS-43-126 was not cytotoxic to Hep3B cells at concentrations up to 100μM, while the LC50 for JS-K was about 10μM. To examine the molecular mechanisms of antitumor effects of JS-K, Hep3B cells were treated with 1-10μM of JS-K for 24h, and then subjected to gene expression analysis via real time RT-PCR and protein immunostain via confocal images. JS-K is a GST-α targeting NO prodrug, and decreased immunostaining for GST-α was associated with JS-K treatment. JS-K activated apoptosis pathways in Hep3B cells, including induction of caspase-3, caspase-9, Bax, TNF-α, and IL-1β, and immunostaining for caspase-3 was intensified. The expressions of thrombospondin-1 (TSP-1) and the tissue inhibitors of metalloproteinase-1 (TIMP-1) were increased by JS-K at both transcript and protein levels. JS-K treatment also increased the expression of differentiation-related genes CD14 and CD11b, and depressed the expression of c-myc in Hep3B cells. Thus, multiple molecular events appear to be associated with anticancer effects of JS-K in human hepatoma Hep3B cells, including activation of genes related to apoptosis and induction of genes involved in antiangiogenesis and tumor cell migration.

Inhibition of Mahkota Dewa (Phaleria macrocarpa) bioactive fraction on proliferation of human retinoblastoma tumor cells Y-79 through suppression of mRNA level of cyclin E

ObjectiveTo prove the molecular mechanisms of Mahkota Dewa (Phaleria macrocarpa) in suppressing proliferation of human retinoblastoma cells through suppression of cell cycle's gene-regulators expression. MethodsIn this study, the molecular mechanism of anti-tumor effect of fractioned extract of Phaleria macrocarpa (DLBS1425) in human retinoblastoma cells Y-79 was investigated by measuring the tumor cells viability, the assessment of population profiles of tumor cells in the cell cycle, and the mRNA concentration of p16, p21, p53, cyclin D, cyclin E, and E2F. ResultsDLBS1425 showed an inhibition effects towards proliferation of Y-79 cell line. Inhibition of proliferation was shown by suppression of cell cycle progression. DLBS1425 downregulated cyclin E, a G1 phase regulator gene of cell cycle, in dose-dependent manner without affecting p53–p21 pathway. In the other word, DLBS1425 inhibits cell proliferation through suppression of cyclin E independently towards conventional proliferation pathway. ConclusionsOur results suggest that DLBS1425 is a potential anticancer agent which targets genes involved in cell proliferation in human retinoblastoma cells which make it pharmacologically ideal for the prevention and/or treatment of retinoblastoma cancer.

O2-2-5 - Clinical verification of a molecular mechanism of antitumor effect of eribulin chemotherapy

O2-2-5 - Clinical verification of a molecular mechanism of antitumor effect of eribulin chemotherapy

Abstract 3535: A novel tumor-specific oncolytic biological therapy against invasive pancreatic cancer

Abstract Pancreatic cancer is one of the worst prognosis disease all over the world and has an overall 5-year survival rate of less than 5%. At the time of diagnosis, more than 80% of patients are not eligible for curative surgical resection due to extensive local tumor invasion and early systemic metastasis. Moreover, even after curative surgery, pancreatic cancers still show poor prognosis due to local recurrence and systemic metastasis. Gemcitabine is the standard chemotherapeutic agent to treat advanced pancreatic cancers, but median overall survival ranged from 5.0 to 7.2 months, suggesting that standard chemotherapy is less effective to most of pancreatic cancer patients. Therefore, the development of novel therapeutic strategy is needed for the treatment of pancreatic cancers. We developed a novel oncolytic adenovirus, OBP-702, in which a human telomerase reverse transcriptase gene promoter drives viral E1 gene for virus replication, and armed with the wild-type tumor suppressor p53 gene. Precise antitumor effects of OBP-702 have been evaluated in human pancreatic cancer cells. We used four pancreatic cancer cell lines with different invasion ability, including non-invasive type (MIA PaCa-2, Panc-1) and invasive type (BxPC-3, Capan-1). The antitumor effect of OBP-702 for each cell line was assessed using XTT assay. OBP-702 induced profound anti-tumor effect in all human pancreatic cancer cells, whereas OBP-301 lacking the p53 tumor suppressor gene showed differential effects among the cell types. OBP-301 induced moderate anti-tumor effect in MIA Paca-2, BxPC-3 and Capan-1 cells and strong anti-tumor effect in Panc-1 cells in a dose-dependent manner, suggesting the broad spectrum of OBP-702's efficacy. The molecular mechanism of anti-tumor effect was assessed by Western blot analysis. OBP-301 induced autophagy-related cell death, whereas OBP-702 induced autophagy- and apoptosis-related cell deaths. These results suggest that an oncolytic adenovirus, OBP-702, is a promising antitumor agent to induce profound cell death in invasive human pancreatic cancer cells. The clinical trial of intratumoral administration of OBP-702 in patients with invasive pancreatic cancer is warranted. Citation Format: Takeshi Koujima, Hiroshi Tazawa, Naoto Hori, Shuta Tamura, Shinji Kuroda, Hiroyuki Kishimoto, Masahiko Nishizaki, Yasuo Urata, Shunsuke Kagawa, Toshiyoshi Fujiwara. A novel tumor-specific oncolytic biological therapy against invasive pancreatic cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3535. doi:10.1158/1538-7445.AM2015-3535

Fascaplysin sensitizes cells to TRAIL-induced apoptosis through upregulating DR5 expression

This study investigated the molecular mechanism of anti-tumor effect of fascaplysin, a nitrogenous red pigment firstly isolated from a marine sponge. Microarray analysis show that the TNF and TNF receptor superfamily in human umbilical vein endothelial cells (HUVEC) and human hepatocarcinoma cells (BEL-7402) were significantly regulated by fascaplysin. Western Blot results reveal that fascaplysin increased the expression of cleaved caspase-9, active caspase-3, and decreased the level of procaspase-8 and Bid. Flow cytometry and cytotoxicity tests indicate that fascaplysin sensitized cells to tumor necrosis-related apoptosisinducing ligand-(TRAIL) induced apoptosis, which was markedly blocked by TRAIL R2/Fc chimera, a dominant negative form of TRAIL receptor DR5. Therefore, our results demonstrate that fascaplysin promotes apoptosis through the activation of TRAIL signaling pathway by upregulating DR5 expression.

Molecular Mechanism of Anti-tumor Effect by Triptolide in Hematological Malignancies

Triptolide is the major active ingredient of extracts derived from the traditional Chinese Medicine tripterygium wilfordii Hook F. As a complex natural extract, triptolide has a broad range of biological target points and exhibits a variety of pharmacological effects including anti-inflammatory, immunosuppressive, antioxidant, and anti-infertility. In addition, considerable evidences show that triptolide holds dramatic anti-tumor activities manifested by antiproliferative and proapoptotic activities at low nanomolar concentration. In clinical trial, some clinical departments in China have used triptolide to treat leukemia without toxicity of heart, liver, kidney as well as gastrointestinal tract at therapeutic dose. The present review delineates the molecular mechanism of the anti-tumor activities of triptolide in hematological malignancies. Our research shows the extensive biological effects of triptolide are mediated by means of perturbing multiple signal pathways and molecular cascades including regulation of tumor cell survival pathway (Bcl-2, Mcl-1, XIAP, caspases, nuclear factor-kappaB ), blocking the progression of cell cycle (CDKs, cyclin D1, P21wap1/cip1 and P27kip1), epigenetic modulation of target genes via interaction with histone methyltransferases and demethylases (EZH2, SUV39H1, NSD1, SMYD3, LSD1, JMJD2B, RIZ1), as well as regulation of angiogenesis pathway (VEGF, VEGFR) in hematological malignancies.

Molecular Mechanism of Anti-tumor Effect by Triptolide in Hematological Malignancies

Molecular Mechanism of Anti-tumor Effect by Triptolide in Hematological Malignancies

Effect of Danshensu on redox state and relevant nuclear transcription factors in non-small cell lung cancer A549 cells

To study the molecular mechanism of anti-tumor effect of Danshensu (DSS) focusing on whether it is related to its antioxidation effect on redox state of non-small cell lung cancer A549 cells and relevant nuclear transcription factors and signal pathway. DPPH radical scavenging, ferric reducing, ferrous iron chelating assay and DCFH-DA fluorescence staining of ROS were introduced to detect antioxidant activity and ROS scavenging activity in vitro; MTf was adopted to analyze the effect of DSS on A549 cells viability at different time and dose. Western blot was used to detect the effect of DSS on the protein expression of hypoxiainducible factor HIF-1alpha and redox regulating transcription factor Nrf2 in A549 cells and the phosphorylation of upstream Akt, ERK1/2 MAPK. DSS could reduce the ROS level in A549 cells in a dose-dependent manner, which was related to it's strong DPPH radical scavenging activity. Moreover, DSS inhibited A549 cells proliferation in both dose- and time- dependent manner. Further study on molecular mechanism indicated that DSS suppressed redox regulator Nrf2, and down-regulated the phosphorylation of Akt and ERK1/2. However, DSS had no effect on HIF-la expression. DSS might have the effect on treating non-small cell lung cancer by scavenging ROS and then inhibiting phosphorylation of Akt, ERK1/2 and down-regulating Nrf2 expression in A549 cells.

Proteomic analysis of anti-tumor effects by tetrandrine treatment in HepG2 cells

Tetrandrine (TET), a bis-benzylisoquinoline alkaloid isolated from the root of Hang-Fang-Chi (Stephenia tetrandra S Moore), exhibits broad pharmacological effects, including anti-tumor activity. Recently, the beneficial effects of TET on cytotoxicity towards tumor cells, radiosensitization, circumventing multidrug resistance, normal tissue radioprotection, and antiangiogenesis have been examined extensively. To explore the potential molecular mechanism of the anti-tumor effect of TET, we applied proteomic tools to profile the proteins in HepG2 cells subjected to TET treatment. The levels of 39 proteins in cells exposed to TET (IC₅₀=5±0.6 μg/ml) for 48 h were observed to undergo significant alterations. Six proteins were identified by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) using peptide fingerprinting from 10 protein spots (density difference >1.5-fold between the control and TET-treated group). Among them, 5 proteins were downregulated (proteasome activator complex subunit 3, 40S ribosomal protein S12, phosphoglycerate mutase 1, destrin, transaldolase) and 1 protein was upregulated (guanylate kinase 1) by TET treatment in HepG2 cells as determined by spot volume (P<0.05). Most of the identified proteins were associated with tumor growth, migration, and anti-tumor drug resistance. These data will be helpful in elucidating the molecular mechanism of TET's anti-tumor effect in HepG2 cells.

ANTITUMOR EFFECT OF SAFFRON (CROCUS SATIVUS L.): OVERVIEW AND PERSPECTIVES

From ancient times, saffron dried stigmas of Crocus sativus L. is widely used as a drug against different human diseases. In the beginning of 1990´s scientific literature reported for the first time the antitumor activity of saffron. During the last decade different laboratories of the world (including ours) have accumulated sufficient scientific evidence to suggest that saffron, and its main ingredients, can inhibit the process of carcinogenesis in vivo and in vitro effectively. The aim of this work has been to summarize and discuss the scientific results obtained in our laboratory, together with data collected from the literature, about the antitumor and anticarcinogenic activities of saffron and its main ingredients, and the possible molecular mechanism of these actions. Saffron and its carotenoid ingredients are chemopreventive in cultivate human malignant cells and animal models, inhibiting cell growth. This could explain how they reduce outgrowth of tumor cells in vivo. It has been shown that those malignant cells are more sensitive than normal cells to the inhibitory effect of saffron. Inhibition of intracellular nucleic acid synthesis and free radical chain reactions may contribute to explain the molecular mechanism of antitumor effect of saffron. Although, it may be too soon to celebrate, the antitumor activity of saffron is quite promising and warrants further investigations, especially in clinical trials.