Alkaloids ; pharmacology ; Cisplatin ; pharmacology ; Fluorouracil ; pharmacology ; Hep G2 Cells ; Humans ; Quinolizines ; pharmacology ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; metabolism

In an attempt to develop a new anticancer platinum complex with greater or equivalent antitumor activity but reduced side effects compared with cisplatin (CDDP), a series of new platinum complexes having a glycolate leaving ligand was synthesized. Among them, five complexes were selected for further development on the basis of adequate water solubility, low nephrotoxicity and high antitumor activity in a murine system. The chemosensitivity of these five complexes was examined in MTT assay against two human pulmonary adenocarcinoma cell lines, PC-9 and PC-14, and two human stomach adenocarcinoma cell lines, MKN-45 and KATO III. Their IC50 and relative antitumor activity (RAA) values were compared with those of CDDP and 254-S, a second-generation platinum complex with a glycolate leaving ligand under phase III clinical trial. The lowest mean IC50 value was observed in CDDP, followed by SKI 2034R and SKI 2033R. In this study, the antitumor activity was evaluated in terms of RAA values and SKI 2034R showed the highest RAA value. The order of RAA values was SKI 2034R > CDDP > SKI 2032R > SKI 2033R > SKI 2030R > SKI 2029R > 254-S. Based on the RAA order, we have recommended SKI 2034R as the most promising candidate for further development of a clinically useful platinum complex.
Antineoplastic Agents/*pharmacology ; Cisplatin/pharmacology ; Comparative Study ; Drug Evaluation, Preclinical ; Human ; Organoplatinum Compounds/*pharmacology ; Tumor Cells, Cultured

Chemotherapy as a routine method for clinical treatment of cancer has disadvantages such as significant toxicity and strong resistance. In order to improve the efficacy of the drugs and reduce the by-effects, we tried to combine static magnetic field (SMF) with cisplatin or adriamycin. The growth of Hepa1-6 cells treated with the static magnetic field (SMF) combined with cisplatin or adriamycin was significantly inhibited, as detected with MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) test. Combined treatment group cells underwent significant morphological changes as observed by HE (Hematoxylin and eosin) staining under optical microscope. Cell cycle analysis indicated that SMF increased the ratio of cells arrested in G2/M phase caused by cisplatin, and when treated with SMF combined with adriamycin, cells were almost arrested in G1 and G2/M phase. SCGE test showed that SMF can enhance the ability of cisplatin or adriamycin to promote cell DNA damage. Atomic force microscope observation found that the combination of antitumor drugs and magnetic field treatment induced larger and deeper holes on the cell membrane, and surface structure damage is serious. The combination of antitumor drugs and magnetic field technology effectively inhibits the growth of tumor cells, and reduces drug doses. The results implicate this method as potential cancer therapy.
Antineoplastic Agents ; pharmacology ; Cell Cycle ; Cell Line, Tumor ; drug effects ; Cisplatin ; pharmacology ; DNA Damage ; Doxorubicin ; pharmacology ; Humans ; Magnetic Fields

Cisplatin (CDDP), homoharringtonine (HHT), mitoxantrone (MIT) and hydroxycamptothecin (HCPT) are highly effective anti-tumor drugs. To evaluate their effects in the therapy of leukemia and establish a valuable method to estimate anti-tumor drugs, Annexin V/PI double parameter flow cytometry was used to detect the effects of these drug inducing apoptosis and death in Jurkat cell line. The results showed that MIT and HCTP-induced apoptosis effects on Jurkat cell line were obvious at 4 hours in early phase after adding drug (P < 0.05) and at 8 hours in late phase after adding drug (P < 0.05). HHT had obvious effect on inducing apoptosis of Jurkat cells, but no significant difference from low to high doses. The effect of CDDP on inducing apoptosis of Jurkat cell line was obviously weaker than that of HHT, MIT and HCPT, its weak effect on apoptosis of Jurkat cell line was found only at high concentration of drug for long time. Death effects on Jurkat cell line can not be observed in every experimental group. It is concluded that low dose of MIT can effectively induced apoptosis of Jurkat cell line. Annexin V/PI double parameter flow cytometry can be used as a reliable method for clinical screening anti-tumor drugs.
Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Camptothecin ; pharmacology ; Cisplatin ; pharmacology ; Drug Screening Assays, Antitumor ; methods ; Harringtonines ; pharmacology ; Humans ; Jurkat Cells ; Mitoxantrone ; pharmacology

OBJECTIVETo assess the protection against cisplatin-induced ototoxicity by adenovirus-mediated overexpression of the bcl-2 gene in cultured spiral ganglion cells (SGC).

METHODSSGC from P3 rats were cultured in vitro and exposed to adenovirus vector carrying green fluorescent protein gene (Ad-GFP), followed by immunocytochemical analysis for expression of the neuron-specific marker Neurofilament 200 (NF200) and detection under laser scanning confocal fluorescence microscope. Then, SGC were transduced by Ad-bcl-2 and the expression of human bcl-2 protein was evaluated by Western Blot. Finally, the cultures of SGC were divided into 4 groups: the group of Ad-bcl-2 transfection followed by cisplatin treatment, the group of Ad-GFP transfection followed by cisplatin treatment, the group of cisplatin treatment only and the untreated group. Cisplatin worked for 48 hours at a concentration of 2 microg/ml. Outcome measures included survival number of SGC and longest neurite length by using ImageJ software.

RESULTSSGC were cultured successfully in vitro and transfected by adenovirus vector safely and efficiently. By Western Blot, human bcl-2 protein was expressed in the group after exposure to Ad-bcl-2, but not in the Ad-GFP transfected SGC. Cisplatin exposure resulted in shrinking of neuritis and pyknosis of cell body, even cell death. Expression of bcl-2 in the SGC provided a significant level of protection against cisplatin-induced SGC degeneration.

CONCLUSIONSOur results suggest that SGC can be transduced by adenovirus vector safely and efficiently in vitro. Adenovirus-mediated delivery of the bcl-2 gene attenuates cisplatin-induced SGC degeneration.

Adenoviridae ; genetics ; Animals ; Apoptosis ; drug effects ; Cisplatin ; pharmacology ; Genes, bcl-2 ; Humans ; Spiral Ganglion ; cytology

OBJECTIVETo compare the differences in antitumor activity between cisplatin (CDDP)-loaded liposomes and nanoparticles in vitro.

METHODSCDDP-gelatin nanoparticles (GPs-Pt) and CDDP-liposomes with similar size, zeta potential, drug loading efficiency and in vitro release property were prepared. The uptake in A549 cells and elimination kinetics were evaluated and antitumor activity was determined by MTT test. The internalization pathways of nanocarriers were studied with inhibitors.

RESULTSInternalization of two nanocarriers was clathrin and actin dependent. Pt accumulation delivered by GPs-Pt was significantly higher than that of liposomes. However, the results of kinetic analysis showed that liposomes had longer cellular retention, and the MRT and AUC were 3 times and twice of GPs-Pt, respectively. The IC(50) of liposomes was significantly lower than GPs-Pt. The values were 2.94±0.21 and 20.70±1.05 μg/ml, respectively.

CONCLUSIONNanocarriers with similar pharmaceutical parameters can induce differences in cellular internalization and elimination, which influence the antitumor activity eventually. Compared with gelatin nanoparticle, liposome is preferable for cisplatin delivery.

Adenocarcinoma ; pathology ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cisplatin ; pharmacokinetics ; pharmacology ; Drug Carriers ; Humans ; Liposomes ; Lung Neoplasms ; pathology ; Nanoparticles

The study is to report the establishment of a method of screening the antitumor compounds based on the dynamic bio-response profile of cells to make up for the shortages of conventional end-point tests such as tedious operation and low sensitivity. Based on the principle of electric impedance of cells, the real-time cell electronic sensing (RT-CES) system was used to monitor the effect of epirubicin (EPI), cisplatinum (DDP) and carboplatin (CBP) on the growth of HepG2 cells, with the cell index (CI), half maximal inhibitory concentration (IC50) and detachment curve as evaluation indexes. Meanwhile, cell counting kit-8 (CCK-8) and microscopy were applied for verification. The results showed that CI curve could sensitively real-time profile the inhibitory effect of model drugs on HepG2 cells. The IC50 of EPI, DDP and CBP were 0.53 +/- 0.04, 9.79 +/- 0.26 and 597.00 +/- 3.79 microg x mL(-1), respectively. What's more, the significant differences of detachment curves of the three drugs indicated that their functional mechanisms might be different, this is consistent with the literature. The RT-CES system with non-invasive, label-free and real-time characteristics could be used to monitor the bio-response profile of the three drugs to HepG2 cells, allowing to qualitatively and quantitatively distinguish the antitumor activities of the three drugs, and could be a complementary method for the present screening of antitumor compounds.
Antineoplastic Agents ; pharmacology ; Biosensing Techniques ; methods ; Cell Count ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Drug Screening Assays, Antitumor ; Electric Impedance ; Humans

BACKGROUNDCisplatin based chemotherapy is a well recognized risk factor for coagulation disorders and thrombosis. The pathophysiological mechanisms by which cisplatin promote thrombosis are not well understood.

METHODSRed blood cells (RBCs) were separated from peripheral blood of patients with breast cancer (n = 10) and healthy adults (n = 6) and treated with cisplatin. Coagulation time of RBCs was assessed by one step recalcification time and the productions of thrombin, intrinsic and extrinsic factor Xa were measured in the presence or absence of various concentrations of lactadherin. Exposed phosphatidylserine was stained with lactadherin and observed by confocal microscopy and flow cytometry.

RESULTSNeither fresh RBCs nor RBCs treated without cisplatin had potent procoagulant activity. Cisplatin treatment increased procoagulant activity of RBCs in a cell number- and concentration-dependent manner. Exposed phosphatidylserine was stained with lactadherin and after cisplatin treatment, strong fluorescence was revealed by confocal microscopy. Lactadherin bound RBCs from patients with breast cancer increased from (1.9 +/- 0.5)% on control RBCs to (68.0 +/- 3.5)% on RBCs treated with 10 micromol/L cisplatin for 24 hours.

CONCLUSIONSCisplatin treatment increases procoagulant activity of RBCs, which have a strong association with exposure of phosphatidylserine. The increased procoagulant activity may contribute to the pathogenesis of thrombophilia during cisplatin based chemotherapy in breast cancer patients.

Antineoplastic Agents ; pharmacology ; Blood Coagulation ; drug effects ; physiology ; Cisplatin ; pharmacology ; Erythrocytes ; drug effects ; physiology ; Humans ; In Vitro Techniques

OBJECTIVETo establish an in vitro model of mouse cochlear basilar membrane impairment using cisplatin, and observe the protective effect of methionine on the hair cells.

METHODSThe cochlear basilar membrane samples of thirty two Kunming mice were harvested on the 2nd day after birth and randomly divided into four groups. Each group had 16 samples. Overnight preincubation the cochlear organ followed by appropriate treatment respectively as follows: the serum-free culture medium, the serum-free culture medium with methionine and cisplatin, the cisplatinum-containing serum-free culture medium, and the methionine-containing serum-free culture medium. The protective effect of methionine for injury of cochlea hair cells induced by cisplatin was observed by myosin-VI immunofluorescence, light microscopy, laser confocal scanning microscope and hair cells counting.

RESULTSThe outer hair cells (OHC) and inner hair cells (IHC) of control group and methionine group were not damaged. The outer and inner hair cells of cisplatin group were damaged in various degree, and had remarkable difference compared with control group and methionine group (P < 0.05). The outer hair cells and inner hair cells of cisplatin + methionine group were damaged less than the cisplatin group with remarkable difference (t(IHC) = 3.929, t(OHC) = 8.582, P < 0.05).

CONCLUSIONSCisplatinum could damage the cochlear hair cells of the basal membrane in Kunming mice. Methionine might protect against cisplatin's damage on the cochlear hair cells.

Animals ; Cisplatin ; adverse effects ; pharmacology ; Hair Cells, Auditory ; drug effects ; In Vitro Techniques ; Methionine ; pharmacology ; Mice ; Mice, Inbred Strains

OBJECTIVE: To establish a CDDP-resistant cell line from human nasopharyngeal carcinoma and evaluate its biological characteristics. METHOD: By continuously exposing and gradually increasing dose of cisplatin (CDDP), a resistant nasopharyngeal carcinoma cell line (HNE1/CDDP) was established. Drug sensitivity of this cell line was detected by MTT assay; the alterations of its biological characteristics were determined using light microscopy, cell counting and flow cytometry (FCM); its ability of adhesion, migration and invasion were also evaluated. RESULT: HNE1/CDDP cell line was developed after 10 months with stable resistance to cisplatin with the resistance index was 5.83. HNE1/CDDP cell exhibited cross-resistance to many other chemotherapeutic agents (carboplatin, oxaliplatin and etoposide, etc). The morphology of HNE1/CDDP changed; doubling time prolonged; and the cell number of S-phase and G2/M-phase decreased while of G0/G1 phase increased compared with parental cells. The ability of adhesion, migration and invasion had no difference between the parental and the resistant cells. CONCLUSION HNE1/CDDP cell line shows the typical and stable resistant phenotype and can be used as a research model.
Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Humans ; Nasopharyngeal Neoplasms