Blocking of methionine aminopeptidase-2 by TNP-470 induces apoptosis and increases chemosensitivity of cholangiocarcinoma

Context: Resistance of cancer cells to chemotherapeutic drugs is a major pitfall of the failure of chemotherapy treatment for cholangiocarcinoma (CCA). A new therapeutic strategy that can improve treatment efficacy is mandatory for CCA patients. Our previous findings demonstrated the overexpression of methionine aminopeptidase-2 (MetAP2) in CCA patients. In addition, supplementation of TNP-470, a MetAP2 inhibitor, significantly inhibited the growth and metastatic activities of CCA cell lines. However, the molecular mechanism of antitumor activity of TNP-470 and the synergistic antitumor activity of TNP-470 combined with chemotherapeutic drugs are still unknown. Aims: The aim of this study is to evaluate the molecular mechanism of anticancer activity and the potential use of TNP-470 as a chemosensitizing agent in CCA cell lines. Materials and Methods: Cell cycle and apoptosis of CCA cell lines were evaluated using flow cytometry with propidium iodide staining. Expression of apoptosis regulatory proteins was measured by Western blotting. The chemosensitizing effect of TNP-470 was determined using combination index. Results: TNP-470 inhibited the growth of CCA cells via induction of apoptosis through activation of the p38-phosphorylation and up- and down-regulation of Bax and Bcl-xL, respectively. Furthermore, TNP-470 significantly enhanced the antitumor activity of 5-fluorouracil, cisplatin, doxorubicin, and gemcitabine. Conclusions: The present results show that TNP-470 could be a potential therapeutic or adjuvant agent for CCA

Experimental studies of angiocytotoxic therapy against growth and metastasis of pancreatic carcinoma / 中华消化杂志

Objective The anti tumor and anti metastasis effects of angiocytotoxic therapy (TNP 470/Gemcitabine) were investigated using a model of human pancreatic carcinoma by surgical orthotopic implantation (SOI). Methods The SOI model was developed by suturing small pieces of SW1990 tumors into the tail of pancreas in nude mice. Twenty four male mice were randomly divided into control group, G100 group receiving 100 mg/kg Gemcitabine intraperitoneally injection on days 0,3,6 and 9 after transplantation, and T30 group receiving 30 mg/kg TNP 470 subcutaneous injection on alternate days for 8 weeks. Another thirty two male mice were randomly divided into control group, T15 group, G50 group and combination group (TNP 470 30 mg/kg+ Gemcitabine 50 mg/kg). Animals were sacrificed ten weeks after transplantation. Results G100 group had a significant inhibitory effect on tumor growth of pancreatic carcinoma compared to T30 group, while the metastasis of tumor was significantly inhibited by T30 group compared to G100 group. Neither G100 group nor T30 group showed a significant improvement on survival rate. T15 group and G50 group alone had no significant inhibitory effect on the tumor growth and its metastasis. Mean while a significant anti tumor, anti metastatic effect and a significant improvement on the survival rate were observed in combination group. The inhibitory effect of G50 group was enhanced by 2 times with T30, and 2/8 of the tumors bearing animals were cured by the combination therapy. The level of microvessel density in T30 group was significantly lower than that in T15 group and control group ( P

The effect of TNP-470 treatment on ascitic-fluid type hepatocarcinoma H22 / 肿瘤研究与临床

Objective To study the effect of TNP-470 treatment on ascitic hepatocarcinoma H22 and potential mechanisms. Methods The experimental approaches employed in the study included in vitro assays of cell proliferation and migration, and in vivo ascitic hepatocarcinoma mouse model. Results In vitro, TNP-470 inhibited cell proliferation of both H22 and human microvascular endothelial cell HMEC. Furthermore, TNP-470 inhibited HMEC migrations. In vivo, TNP-470 treatment significantly improved the life quality of the tumor-loaded mice, including lessen the ascitic effasion, decreased tumor cell viability, lessen the eye blindness, and prolonged the life-span. Conclusion These observations demonstrate that TNP-470 would be an effective drug for treatment of ascitic-fluid type hepatocarcinoma mice.

Synergistic Inhibition of TNP-470 and Recombinant Human Endostatin on the Growth of Mice Lung Adenocarcinoma LA795 / 中国肿瘤生物治疗杂志

Objective: To investigate the combined inhibitive effect of TNP-470 and rhES on the growth of lung adenocarcinoma LA795 in T739 mice. Methods: The purified rhES was acquired by using methanol to induce the recombinant pichia pastoris. GS115 and heparin affinity chromatography. The T739 mice inoculated with LA795 cells were randomized into three groups, 10 mice per group, one group was injected with PBS for 14 days, the other two groups were respectively treated with rhES and TNP-470+rhES. To observe the tumor growth in different groups, and the tumor volume was measured with caliper. The microvessel density(MVD) of tumors were measured by using immunohistochemistry. Results: The purified rhES was acquired. In compared with PBS group, the tumor growth of other two groups was inhibited significantly. And the tumor volume of TNP-470+rhES group are smaller than the rhES group (P

Inhibitory Effect of Liver Metastasis of Human Colon Cancer by TNP-470 in Combination with 5-FU / 中国肿瘤生物治疗杂志

Objective: To study the effect of angiogenesis inhibitor TNP-470 combination with 5-FU on liver metastasis of human colon cancer. Methods: Human colon cancer cell line, LOVO cells, were injected intrasplenically into BALB/c nude mice to produce diffuse liver metastases. Mice were randomly divied into four groups; TNP-470 treated group, 5-FU treated group, TNP-470 +5-FU treated group and control group. Animals were sacrificed after 4 weeks, and their livers were processed for histological examination. Liver metastatic rate and tumor foci in liver were counted. Tumor microvessel density (MVD) and vascular endothelial growth factor (VEGF) were determined by immunohistochemistry SABC method with image analyse system. Results: TNP-470 in combination with 5-FU and TNP-470 alone display a significant inhibitory effect on liver metastasis compared to the control ( P

Effects of TNP-470 on proliferation, the cell cycle and apoptosis in human colon cancer Lovo cells / 中国药理学通报

AIM To study the effect of TNP 470 on proliferation, cell cycle and apoptosis in the cultured human colon cancer Lovo cells. METHODS The growth inhibition of TNP 470 on Lovo cells was evaluated by an MTT.assay The effect of TNP 470 on cell cycle and apoptosis measured by flow cytometry, and ultrastructural feature of Lovo cells were observed with electromicroscope. RESULTS TNP 470 inhibited the growth of Lovo cells. flow cytometry analysis showed that G 0/G 1 phase rate was increased but S phase rate was decreased. Apoptosis rate of TNP 470 treated group was significant high than that of control and typical chang of apoptosis in Lovo cells was observed. CONCLUSION TNP 470 can inhibit proliferation of Lovo cells, and this inhibition is associated with cell cycle block and apoptosis.