Anticancer Effects And Mechanisms Of Chiral Ruthenium Complexes Λ-WH0402

Cisplatin is one of the world's three major anticancer drugs, whose wide use has led to a positive recognition of metal antitumor drugs. Ruthenium complexes, which have the same chemical structure and properties as platinum are considered as one of the most promising metal anti-cancer drugs. Because of the serious side effects of cisplatin, researchers started to focus on metallic ruthenium complexes which have similar anti-tumor activity as platinum.In recent years, most studies have found that ruthenium complexes are of low toxicity, easily digestible, and can be quickly discharged, and that its pharmacological activity tends to increase owing to the synergy of ligand and metal ions. The high concentration of these compounds in the tumor shows they are easily absorbable by tumor tissues, and unlike platinum compounds, have better biological activity and lower toxicity. Therefore, ruthenium complexes, which may be more active than cisplatin, promise to become the most promising anti-cancer drugs given appropriate chemical modifications.The first stage of the project involved obtaining a series of chiral ruthenium bipyridine and phenanthroline chiral ruthenium complexes through the design and synthesis of different ligands and changing ancillary ligands, and, and then characterizing these compounds. A comparison of∧-isomers andΔ-isomers found the former to be more active. After a series of screening, ruthenium complex∧-[Ru (bpy) 2 (o-tFPIP)] (Cl) 2, i.e.∧-WH0402, was found to have the best activity among a number of chiral ruthenium complexes.The current research is inter-disciplinary study involving cell biology, experimental zoology, molecular biology, pathology, and so on. It examines the anti-tumor activity of chiral ruthenium complexes∧-WH0402, and discusses its anti-tumor mechanism through screening sensitive cancer strains of the∧-WH0402, and through the experiments of anti-tumor activity of sensitive strains, of anti-tumor metastasis experiment, of inhibition of vascular experiments to study. It does so also by using cisplatin as the control.Method1. Effort was made to determine whether it is necessary to study the∧-WH0402 through screening by MTT on its sensitive tumor cell lines with cisplatin as the control, and through the experiments in vivo and in vitro of testing and verifying the efficacy and toxicity of∧-WH0402.2. Second, effort was made to observe the inhibition of the growth for the strain sensitive tumor cell lines of∧-WH0402 by cell climbing film, electronic microscopy, the cell cycle, and experiments of nude mice in vivo. Then experiments in flow cytometry, fluorescence microscopy, apoptosis protein Westerblot were conducted to observe the apoptosis on sensitive tumor lines whice induced by∧-WH0402. All this helped to observe the anti-tumor activity effected by∧-WH0402 to strain sensitive tumor cell lines.3. HCCLM6 is the sensitive cancer strain of∧-WH0402, for it is the hepatic carcinoma cell line with highly metastatic characteristics. Efforts were made to observe the effects of∧-WH0402 on tumor metastasis through invasion and motility, adhesion experiments in vitro, and the experiment of transplanting in situ in vivo were be achieved.4. Attempt was made to observe the effect of∧-WH0402 on the blood vessels, through human umbilical vein endothelial cells HUVEC, through MTT experiment, endothelial cell migration assay, and experimental hematogenous lung metastasis test in vivo. Results1. Through screening 10 human tumor cell lines by MTT test in vitro, it was shown that the hepatic carcinoma cell lines HCCLM6 with highly metastatic characteristics are the most sensitive cancer strain of∧-WH0402.2. MTT experiments in vivo and nude mice and C57 mice experiments in vitro indicated that the efficacy of∧-WH0402 is similar to that of cisplatin, with no significant difference between sex (P=0.028), and acute toxicity results showed that the LD50 of∧-WH0402 is 11.66 mg/kg.3. Inverted microscope, cells crawl films, scanning electronic microscopy showed that the HCCLM6 cells of the control group grow in good condition, while those in the experimental group gradually deteriorated and growth was increasingly suppressed with increased dosage or incubation time by∧-WH0402. Cell cycle distribution test by flow cytometry showed that at a concentration 0.312μg/ml for 24h, 48h, the cell cycle mainly remained in G0/G1 phase, and when the concentration reached 0.625 and 1.25μg/ml for 24h,48h, the cell cycle mainly remained in the S phase. Flow cytometry, fluorescence staining, westerblot analysis show that∧-WH0402 can induce HCCLM6 cell to apoptosis.4. Nude mice experiments in vivo was conducted where the mice was inoculated with HCCLM6 cell in the right flanks subcutaneously by 1×106, and inoculated with 2×105 in the left flanks. After the establishment of bilateral tumor mouse model, saline was subcutaneously injected in the left tumor on the next day while water-soluble∧-WH0402 was injected in the right tumor. Two-way analysis of variance shows that there was significant difference (F=20.242, P=0.001) in terms of transplanted tumor volume between∧-WH0402 treatment group and the group, and independent samples T test showed that there was significant difference (t= 9.363, P=0.000) in terms of tumor weight between the two groups.5. Invasion, adhesion and movement experimental in vitro showed that A-WH0402 is able to inhibit the invasion, adhesion and movement capability of HCCLM6 cell in vitro. And orthotropic transplantation experiment showed that∧-WH0402 can significantly inhibit the metastasis situation of liver, lung, abdomen, abdominal wall after orthotropic transplantation.6. MTT test of Endothelial cells showed that,∧-WH0402 on endothelial cells is more sensitive than the sensitive strain HCCLM6 cells while endothelial cell migration experiments show that∧-WH0402 can inhibit endothelial cell migration. Additionally, mice with experimental hematogenous lung metastasis experiment showed that the animal numbers of lung metastasis (χ2=4.000 P=0.046), lung metastasis, lung metastasis nodes (t=-6.357 P=0.000) of∧-WH0402 treatment group decreased more markedly than that of the control group.Conclusions1. After screening 10 human tumor cell lines, it was found that the most sensitive tumor cell lines in HCCLM6 is∧-WH0402, which is Hepatic carcinoma with highly metastatic feature.Compare with cisplatin,the toxicity and anticancer activity of A-WH0402 is similar to that of cisplatin.2.∧-WH0402 can inhibit the growth of HCCLM6 cell lines and induce apoptosis.3.∧-WH0402 can inhibit the metastasis capability of HCCLM6 cell lines in vivo and in vitro.4. HCCLM6 cell lines are more sensitive than endothelial cells HUVEC to A-WH0402, which can inhibit the migration of endothelial cells in vitro and the experimental hematogenous metastasis in vivo.The Originality of This StudyRuthenium complexes are used directly for screening of ten human tumor cell lines, which were found to have different inhibitory effect, among the ten tumor cell lines hepatic carcinoma cells HCCLM6 with highly metastatic feather is found to be the most sensitive strains for∧-WH040. The anti-tumor mechanisms depend on promoting the apoptosis and inhibiting the proliferation of tumor cell, blocking angiogenesis and inhibiting invasion and metastasis to play a role in prohibiting cancer cell growth and expansion. The overall potency and side effects of∧-WH040 is close to cisplatin, so that it is deserved to conduct the pre-clinical project.