OBJECTIVETo evaluate the underlying mechanism of Jianpi Jiedu Recipe (, JJR) in the reversion of multidrug resistance concerning colorectal cancer in vitro and in vivo.

METHODSMice were treated orally with JJR at a daily 4.25 g/(kg·day) or injected with vinblastine (VCR) 2.5 mg/(kg·day) for 3 weeks after having been inoculated with HCT8/V cells; tumor tissues were assayed by hematoxylin and eosin staining. Firstly, the effects of JJR on the expression of cyclooxygenase-2 (COX-2) were tested by real-time polymerase chain reaction (PCR) technique and COX-2 gene silenced by siRNA. Secondly, the variation of intracellular concentration of oxaliplatin (L-OHP) was evaluated by the inductively coupled plasma mass spectroscopy (ICPMS) in HCT8/V and its COX-2 siRNA cells; the concentration of JJR combined with chemotherapeutic drugs and the reverse effect of multidrug resistance (MDR) in HCT8/V cells was evaluated by the MTT assay. Thirdly, real-time quantitative PCR and Western blot analysis were used to detect the multidrug resistance gene 1 (MDR1) mRNA and P-gp expression.

RESULTSJJR had an inhibitory effect on the growth of tumors in vivo, and it, in combination with chemotherapeutic drugs, could reverse the drug-resistance of HCT8/V cells and increase the sensitivity of HCT8/V cells to VCR, DDP, 5-Fu, and THP. ICP-MS results showed that JJR could increase the concentration of drugs in HCT8/V cells (P<0.01). Furthermore, it was shown that JJR could reverse drug resistance of colorectal cancer cells by decreasing MDR1 expression and P-gp level via downregulation of COX-2, which has been represented as one of the major mechanisms that contributes to the MDR phenotype (P<0.01).

CONCLUSIONJJR reversed multidrug resistance and enhanced the sensitivity to chemotherapy, which could be attributed to the down-regulation of COX-2 in MDR1/P-gp-mediated MDR colorectal cancer after chemotherapy.

ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; drug therapy ; enzymology ; pathology ; Cyclooxygenase 2 ; genetics ; metabolism ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Green Fluorescent Proteins ; metabolism ; Humans ; Intracellular Space ; metabolism ; Mice, Inbred BALB C ; Organoplatinum Compounds ; metabolism ; RNA, Small Interfering ; metabolism ; Signal Transduction ; drug effects ; Vinblastine ; pharmacology ; therapeutic use ; Xenograft Model Antitumor Assays

Vinflunine tartrate-loaded liposomes (VT-L) with two drug-to-lipid ratios were prepared by pH gradient method. Vesicle size and zeta potential were determined by the Zetasizer Nano ZS. Entrapment efficiency was evaluated by cation exchange resin centrifugalization method. The toxicity and tumor inhibition to nude mouse administrated by VT-L with different drug-to-lipid ratios were investigated and compared with the vinflunine tartrate injection (VT-I). The results showed that the mean particle size, zeta potential and entrapment efficiency of the VT-L with drug-to-lipid ratios of 1 : 5 and 1 : 10 were 124.6 nm and 128.3 nm, -25.3 mV and -22.8 mV, 94.46% and 97.31%, respectively. The VT-L with two different drug-to-lipid ratios has significantly higher anti-tumor effect to nude mouse transplanted human non-small cell lung carcinoma A549 and lower toxicity than VT-I. While there were no significant differences in anti-tumor effect and toxicity between VT-L with two different drug-to-lipid ratios.
Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; chemistry ; pharmacology ; toxicity ; Cell Line, Tumor ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Drug Stability ; Female ; Humans ; Liposomes ; Lung Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Particle Size ; Random Allocation ; Tartrates ; administration & dosage ; chemistry ; pharmacology ; toxicity ; Tumor Burden ; drug effects ; Vinblastine ; administration & dosage ; analogs & derivatives ; chemistry ; pharmacology ; toxicity

The massive reorganization of microtubule network involves in transcriptional regulation of several genes by controlling transcriptional factor, nuclear factor-kappa B (NF-kappaB) activity. The exact molecular mechanism by which microtubule rearrangement leads to NF-kappaB activation largely remains to be identified. However microtubule disrupting agents may possibly act in synergy or antagonism against apoptotic cell death in response to conventional chemotherapy targeting DNA damage such as adriamycin or comptothecin in cancer cells. Interestingly pretreatment of microtubule disrupting agents (colchicine, vinblastine and nocodazole) was observed to lead to paradoxical suppression of DNA damage-induced NF-kappaB binding activity, even though these could enhance NF-kappaB signaling in the absence of other stimuli. Moreover this suppressed NF-kappaB binding activity subsequently resulted in synergic apoptotic response, as evident by the combination with Adr and low doses of microtubule disrupting agents was able to potentiate the cytotoxic action through caspase-dependent pathway. Taken together, these results suggested that inhibition of microtubule network chemosensitizes the cancer cells to die by apoptosis through suppressing NF-kappaB DNA binding activity. Therefore, our study provided a possible anti-cancer mechanism of microtubule disrupting agent to overcome resistance against to chemotherapy such as DNA damaging agent.
Animals ; Antibiotics, Antineoplastic/therapeutic use ; *Apoptosis ; Caspases/metabolism ; Cell Line ; Colchicine/pharmacology ; DNA/metabolism ; *DNA Damage ; Doxorubicin/therapeutic use ; Humans ; Mice ; Microtubules/chemistry/*drug effects/metabolism ; NF-kappa B/antagonists & inhibitors/*metabolism ; Neoplasms/drug therapy ; Nocodazole/pharmacology ; Protein Binding ; Signal Transduction ; Tubulin Modulators/*pharmacology ; Vinblastine/pharmacology

OBJECTIVEThis experiment aims to study the anti-angiogenic ability of vinorelbine combined with cetuximab in vitro and in vivo.

METHODSHuman lung adenocarcinoma A549 cells were used as control group. Proliferation of human umbilical vein endothelial cells (HUVEC) was assessed by MTT assay. Furthermore, we used Transwell chambers, capillary tube formation and flow cytometry to observe the effects of vinorelbine combined with cetuximab on HUVEC migration, tube formation and cell apoptosis, respectively. In addition, the anti-angiogenic ability of the drugs was checked using chicken chorioallantoic membrane (CAM) model.

RESULTSThe inhibitory rate of HUVEC growth was 25.8%, 39.2%, 54.0% for vinorelbine at the concentration of 0.1 ng/ml, 0.4 ng/ml, and 0.8 ng/ml, respectively; that of 0.25 microg/ml cetuximab was 19.7%, and that of 0.1 ng/ml vinorelbine + 0.25 microg/ml cetuximab, 0.4 ng/ml vinorelbine + 0.25 microg/ml cetuximab and 0.8 ng/ml vinorelbine + 0.25 microg/ml cetuximab was 29.5%, 46.4%, 64.6%, respectively. The inhibitory rates of the drugs at the above mentioned combinations of migration and tube formation of HUVEC were 51.9%, 68.2%, 95.0%, respectively. The inhibitory rate of 0.1 ng/ml + 0.25 microg/ml cetuximab and 0.4 ng/ml vinorelbine + 0.25 microg/ml cetuximab on tube formation of HUVEC was 38.8% and 57.7%, respectively, showing a sub-additive effect, and that of combination of 0.8 ng/ml vinorelbine + 0.25 microg/ml cetuximab was 78.9%, showing a synergistic effect. In addition, the apoptotic rate of HUVEC induced by 0.8 ng/ml vinorelbine + 0.25 microg/ml cetuximab was 59.9%, showing a synergistic effect. The in vivo experiment also showed that the combination of the two drugs had a synergistic anti-angiogenic effect.

CONCLUSIONBoth low dose vinorelbine and cetuximab have an anti-angiogenic effect in vitro and in vivo, and the combination of the two drugs has sub-additive or synergistic inhibitory effect on angiogenesis.

Adenocarcinoma ; blood supply ; pathology ; Angiogenesis Inhibitors ; pharmacology ; Animals ; Antibodies, Monoclonal ; pharmacology ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cetuximab ; Chick Embryo ; Drug Synergism ; Endothelial Cells ; cytology ; Humans ; Lung Neoplasms ; blood supply ; pathology ; Neovascularization, Pathologic ; prevention & control ; Umbilical Veins ; cytology ; Vinblastine ; analogs & derivatives ; pharmacology

OBJECTIVETo study the effect of vinorelbine on apoptosis, telomerase activity and expression of hTERT gene in human lung adenocarcinoma cell line Anip973 cells.

METHODSAnip973 cells were cocultured with Vinorelbine at different concentrations and collected at 24 h, 48 h and 72 h after treatment, respectively. The inhibition rate of cell growth was determined by methyl thiazolyl tetrazolium (MTT) assay to evaluate the effect of vinorelbine. The percentage of apoptosis was detected by flow cytometry. The cellular morphology was observed under inverted microscope and electron microscope. Telomerase activity of Anip973 cells was determined by the method of TRAP-PAGE-silver staining. RT-PCR was used to detect the expression of hTERT mRNA.

RESULTSVinorelbine down-regulated the telomerase activity and expression of hTERT gene mRNA, inhibited the growth of Anip973 cells, and induced cell apoptosis in a time- and concentration-dependent manner. Annexin V-FITC assay showed 0.08 µg/ml NVB group could inhibited cell proliferation in 24 hour, and apoptosis rate was (7.37 ± 0.35)%, RT-PCR detection of hTERT mRNA expression in this group was (57.01 ± 1.71), and they were very significantly different from that in the control group (P < 0.01), but telomerase activity was (6.36 ± 0.06), compared with the control group showed no significant difference(P > 0.05). The change of hTERT mRNA expression is more sensitive than telomerase activity. The apoptosis rate, telomerase activity and hTERT mRNA expression of 0.4 µg/ml group and 2.0 µg/ml group were very significantly different from that in the control group (P < 0.01). The telomerase activity of 2.0 µg/ml at 72 hour group was (1.36 ± 0.27), basically completely inhibited, while the apoptosis rate was (74.87 ± 1.88)%, showed the cell apoptosis rate was in a negative correlation with the down-regulation of the hTERT mRNA (r = -0.96046, P < 0.01).

CONCLUSIONSVinorelbine can induce apoptosis in Anip973 cells, and its mechanism of action is related to telomerase activity. The detection of telomerase activity and expression of hTERT gene is useful in predicting prognosis of patients with lung adenocarcinoma.

Adenocarcinoma ; metabolism ; pathology ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dose-Response Relationship, Drug ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Telomerase ; genetics ; metabolism ; Vinblastine ; administration & dosage ; analogs & derivatives ; pharmacology

BACKGROUND AND OBJECTIVEHuman telomerase reverse transcriptase is the catalytic subunit of telomerase, and its activity is correlated with cell's sensitivity to chemotherapy. The aim of this study is to investigate the differential expression of human telomerase reverse transcriptase (hTERT) mRNA in human lung adenocarcinoma cell line Anip973 and Anip973/NVB, and to observe the correlation between hTERT mRNA and drug-resistance.

METHODSThe real-time fluorescence quantitative RT-PCR was used to detect the change of hTERT mRNA in human lung adenocarcinoma drug-resistant cell Anip973/NVB and parental cell Anip973 treated by NVB.

RESULTSIn the control group, the expression of hTERT mRNA showed no significant difference between drug-resistant cell Anip973/NVB and parental cell Anip973. After been treated by NVB, the expression of hTERT mRNA in parental cell was significantly decreased (P < 0.01), and drug-resistant cell Anip973/ NVB had no evidently variant (P > 0.05). The down-regulated hTERT mRNA in Anip973 cell was higher than that in Anip973/ NVB cell.

CONCLUSIONTelomerase correlates with the drug-resistant cell A973/NVB, and telomerase may be a new target for multi-drug resistant inversion.

Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; genetics ; physiology ; Humans ; Lung Neoplasms ; genetics ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Telomerase ; genetics ; Vinblastine ; analogs & derivatives ; pharmacology

This study was purposed to explore the Rituximab (RTX)-mediated sensitization of B-NHL cell lines to apoptosis induced by Gemcitabine or Navelbine and its possible mechanism. The inhibitory rate of B-NHL cell proliferation was detected by XTT method, the IC₅₀ from Gemcitabine or Navelbine and combination of Gemcitabine or Navelbine with RTX was compared. The expression level of antiapoptotic protein BCL-2 in Daudi, Ramos, Raji and Namalwa cells treated with RTX of 20 μg/ml for 24 hours was detected by Western blot. The results showed that the RTX as a single agent could weakly induce the apoptosis of Daudi, Namalwa, Raji and Ramos lymphoma cell lines, the inhibiting rate of cell proliferation ranged from 3% to 10%; RTX could sensitize significantly the cytotoxity of Gemcitabine or Navelbine in Daudi, Namalwa and Raji cell lines; The expression of BCL-2 in Raji and Namalwa cell lines treated with RTX of 20 microg/ml for 24 hours was down-regulated. It is concluded that RTX can sensitize the cytotoxicity of Gemcitabine or Navelbine to the human lymphoma cell lines which displayed the synergistic effect on Daudi, Namalwa and Raji cell lines. The BCL-2 expression level in Raji and Namalwa cell lines treated by RTX is down-regulated which may be one of the mechanisms sensitizing the cytotoxicity of Gemcitabine or Navelbine.
Antibodies, Monoclonal, Murine-Derived ; pharmacology ; Antineoplastic Agents ; pharmacology ; Cell Line, Tumor ; drug effects ; Cytotoxicity, Immunologic ; drug effects ; Deoxycytidine ; analogs & derivatives ; pharmacology ; Humans ; Lymphoma, T-Cell ; Proto-Oncogene Proteins c-bcl-2 ; Rituximab ; Vinblastine ; analogs & derivatives ; pharmacology

This study was purposed to investigate the biological effect of vinblastine (VLS), usually known as inductor of mitotic arrest, on MOLT-4 of ALL cells and to evaluate its significance. The cell arrest in M phase and/or cell apoptosis were induced by treatment of MOLT-4 cells with 0.05 microg/ml VLS for 0 - 12 hours; the DNA histogram was detected by flow cytometry; the morphological changes of cells were observed by confocal microscopy; the cell cycle distribution, cell apoptosis and morphological changes of cells before and after arrest were analyzed by using arrest increasing rate (AIR), arrest efficiency (AE), apoptosis rate (AR) and morphologic parameters respectively. The results indicated that the cell arrest did not accompanied by significant increase of apoptosis rate; the DNA histogram of cell arrest showed dynamic change of cell cycle in time-dependent manner; the arrest efficiency could be quantified. The cell arrest at M phase was accompanied by cell stack in S phase, the cell proliferation rate dropped after cell arrest occurred. The cells arrested at M phase possessed of characteristic morphologic features in cell mitosis. It is concluded that the vinblastine can solely induce arrest of MOLT-4 cells at M phase. This study provides experimental basis for further investigating the relation of cell cycle arrest to apoptosis, mechanism of checkpoint and development of new anticancer drugs.
Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Division ; drug effects ; Flow Cytometry ; Humans ; Tumor Cells, Cultured ; Vinblastine ; pharmacology

We think the strategy of classic natural product-based drug discovery will be an effective way for us to develop new drugs with independent intellectual property. The strategy includes: to study the molecular mechanism of action of classic natural product with chemical genetics and chemical biology approaches firstly; then establish the proper in vitro bioassay or bioassay system based on its molecular mechanism for their pharmacodynamic evaluation; finally, study their structure-activity, structure-toxicity and structure-ADME properties with medicinal chemistry.
Animals ; Anti-HIV Agents ; chemical synthesis ; pharmacology ; Antineoplastic Agents, Phytogenic ; chemical synthesis ; pharmacology ; Berberine ; analogs & derivatives ; chemical synthesis ; pharmacology ; Biological Products ; chemical synthesis ; chemistry ; pharmacology ; Camptothecin ; analogs & derivatives ; chemical synthesis ; pharmacology ; Drug Discovery ; Humans ; Hypoglycemic Agents ; chemical synthesis ; pharmacology ; Oleanolic Acid ; analogs & derivatives ; chemical synthesis ; pharmacology ; Triterpenes ; chemical synthesis ; pharmacology ; Vinblastine ; analogs & derivatives ; chemical synthesis ; pharmacology

OBJECTIVETo compare effects of vinblastine (VLB) nanoparticles (NPS) and VLB physiologic saline solution on inhibiting glioma cell lines C6 growth and inducting its apoptosis.

METHODGlioma cell lines C6 were respectively treated with 500 micro x L(-1) VLB NPS and VLB physiologic saline solution for 7 days. Amount of cells were counted by blood cell counting chamber. Glioma C6 growth curve was draw according to cells amount. Clone formation rate of glioma C6 was detected after 500 microg x L(-1) VLB NPS and VLB physiologic saline solution incubation for 2 weeks. In addition, the whole morphology of glioma C6 were observed by inverted microscope and inverted fluorescence microscope after 500 microg x L(-1) VLB NPS and VLB physiologic saline solution incubation for 48 h.

RESULTEntrapment of VLB in NPS may significantly inhibit glioma cells C6 growth from 2 to 7 days compared with VLB physiologic saline solution in the same dose (P < 0.05). Clone formation rate of glioma C6 in VLB physiologic saline solution group is 1. 3 times better than VLB NPS. The difference between VLB NPS and VLB physiologic saline solution is significant (P < 0.05). Results of morphology change indicated glioma cells C6 with the VLB NPS treatment were intermediate or end stage, missed structure integrality. Amount of cells was distinctly decreased, and apoptosis cells number was apparently increased compared with VLB physiologic saline solution group.

CONCLUSIONVLB NPS have stronger cytotoxicity to glioma cells line C6 compared with VLB physiologic saline solution in the same dose. NPS may be effective as promising carrier for the transport of VLB into the glioma cells.

Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Glioma ; drug therapy ; physiopathology ; Nanoparticles ; chemistry ; Rats ; Vinblastine ; pharmacology