Development Of Mitochondrial Viscosity Probes To Distinguish Homologous Cancer Cells From Normal Ones And Exploration Of Aggregation Behavior Of JC-1 Derivatives

To extend the life of man,to alleviate the disease of man,both the nature of nature,but also the existence of humanity.Cancer is arguably the most life-threatening disease in today’s society,characterized by rapid disease progression and high mortality.Cell carcinogenesis is the source of cancer,which is a complex process with multiple factors and multiple steps.Therefore,one of the tasks of this paper is to enable the diagnosis and monitoring of cancer.In order to solve this problem,this paper makes full use of the difference of mitochondrial viscosity between cancer cells and normal cells,and develops a fluorescence probe sensitive to viscosity and targeting mitochondria based on torsion intramolecular charge transfer(TICT)mechanism.The probe has weak fluorescence in low viscosity environment and strong fluorescence in high viscosity environment.On the one hand,the probe enables no-wash,highfidelity and rapid imaging of mitochondria.On the other hand,it can detect changes in mitochondrial viscosity during drug induction and apoptosis.Most importantly,it was successfully applied to distinguish homologous human breast cancer cells(MCF-7)from normal cells(MCF-10A)through differences in fluorescence intensity caused by abnormal mitochondrial viscosity in cancer cells.JC-1(5,5’,6,6’-Tetrachloro-1,1’,3,3’-tetraethyl-imidacarbocy anine)is a classical fluorescent reagent for the detection of mitochondrial membrane potential.Its principle is aggregation/dispersion,but there are few reports on the aggregation behavior of JC-1 and its derivatives.In addition,external factors(solvent,ionic strength,temperature)can affect the aggregation of cyanine dyes has been widely known.In this paper,based on some reliable basic scientific data obtained,a new mode of aggregation induction is proposed,that is,multivalent anions can induce the formation of J-aggregates.In this paper,JC-1-OH(introducing-OH to enhance the water solubility of dyes)and JC-C8(extending the side chain to improve the lipid solubility of dyes)were designed and synthesized by regulating the side chain of JC-1.Three molecules with different hydrophilicity were compared,their spectral and particle size properties were tested in detail,and the role of polyvalent anions in the aggregation process was simulated by Gaussian calculation,so as to systematically study the formation of Jaggregation induced by polyvalent anions.This provides an explanation for the rational use of JC-1 and explores a new and broader way of inducing aggregation.In conclusion,this paper has achieved the distinction between homologous human breast cancer cells(MCF-7)and normal cells(MCF-10A)by focusing on the important organelle microenvironment of mitochondrial viscosity.Based on JC-1,the formation of J-aggregates induced by multivalent anions was proposed.These probes and dyes can be used as a powerful tool to provide experimental and theoretical basis for related applications.