Study On The Antitumor Activity Of Ruthenium(Ⅱ)metal Complexes

With the rapid development of the economy and medical level,many diseases have been well controlled,but it is difficult to overcome malignant tumors and the tumors always threaten people’s life safety.In the traditional and new anti-tumor treatment methods,chemical drug therapy has always been irreplaceable with its unique advantages,and related research is constantly expanding and innovating.As the"penicillin in anticancer drugs",the accidental discovery of cisplatin changed the use of nitrogen mustard drugs and folic acid antagonists for tumor treatment at that time,which prompts people to turn their attention to the development of anti-tumor drugs to metal complexes.Although the first,second,and third generations of platinum metal complexes-cisplatin,carboplatin and oxaliplatin-have good cytotoxicity and can effectively inhibit the proliferation of tumor cells,they all have toxic side effects that cannot be ignored.As researchers gradually expanded the scope of research to other metal complexes,many metal complexes with excellent anti-cancer effects were found.Among them,due to the advantages of ruthenium metal complexes such as coordination diversity,high oxidation state stability and low toxicity of normal tissues,it has become an important research object of metal anti-tumor drugs.By altering the precursor or ligand structure of ruthenium metal complexes,the researchers hope to find out the effect of structural changes on the anti-cancer activity of ruthenium metal complexes and related anti-tumor mechanisms.In this thesis,we synthesized two series of seven ruthenium metal complexes by introducing fluorine atoms and chlorine atoms,respectively,and then studied their anti-cancer activity and mechanism by combining in vitro cell experiments and in vivo animal experiments.After purification of ruthenium metal complexes by column chromatography,we performed MTT experiments to detect their cytotoxicity.The results showed that Ru1-Ru7 had good cytotoxicity to most tumor cells,and the IC₅₀ value（unit:μM）of very few complexes reached single digits,and there was no cytotoxicity to normal cells.Subsequently,through cell scratching and cell cloning experiments,we observe the inhibition of complexes on the proliferation and migration of tumor cells.Combined with endocytosis and cell co-localization experiments,it can also be found that complexes can smoothly enter tumor cells and localize at mitochondria and lysosomes.Using flow cytometry to detect the effect of complexes on cell cycle and apoptosis,combined with high-content imaging system to observe the completion of cell division and whether there is autophagy,we found that the complexes have the effect on inducing tumor cell cycle blockage,incomplete division,and significantly increased apoptosis ratio,and there is a phenomenon of autophagy.After the detection of intracellular reactive oxygen species（ROS）content,it was found that complexes led to a significant increase in ROS levels.Then the mitochondrial permeability transition pore（MPTP）and mitochondrial membrane potential（MMP）were detected to find that the complexes caused the tumor cell MPTP to continue to open and MMP to depolarize.In addition,we found that the cytochrome C released from the mitochondria enter the cytoplasm when the complex acted on the mitochondria,which caused a rise in the intracellular ROS content.Western blotting analysis shows that complexes can significantly induce apoptotic and regulate the expression of autophagy-related protein.These proteins mostly activate the Caspase protein family via Bcl-2/Bax protein to induce apoptosis.Furthermore,inhibition of the AKT/mTOR signaling pathway would also result in autophagy occurrence.Finally,antitumor activity in vivo demonstrates that ruthenium metal complexes can effectively inhibit tumor growth.