| At present,cancer has become one of the most serious diseases causing human to death and threatening human health.The traditional means of cancer therapies,such as surgical resection or chemotherapy,have low treatment efficiency as well as high side effects that cannot meet the requirements for the treatment of cancer patients now.Gene therapy,a new therapeutic method for the treatment of cancer,whose characteristics is to deliver nucleic acid polymers into a patient's cells as a drug,has many advantages superiors to traditional ones.However,the current gene therapy vectors used in clinical practices are mostly derived from viral vectors;in which there evoke potential risks,such as immunogenicity,toxicity,or other unknown issues.Therefore,how to secure delivery of the therapeutic gene into the tumor cells and make it effective is the critical bottleneck in the current studies.As a hotspot detection technique,fluorescent probe is of high sensitivity,good selectivity,and membrane permeability.A delivery system will be established via the structure of fluorescent probe to load the treatment gene,and it can also play its own detection capabilities,as well as treat the tumor by delivering the drug to tumor targeting sites.Based on the structure of fluorescent probe,a novel rare earth upconversion luminescence nano-fluorescent probe was constructed to targeted deliver small interfering RNA.The optimal preparation conditions of rare earth nano-particles have been investigated via TEM and fluorescence Spectroscopy.The optimal ratio of particles to RNA was presented through N/P.Finally,PCR and Western Blot were used to detect the delivery efficiency and protein expression of the probe delivery to siRNA.The results show that the morphology,particle size,dispersion,and luminous intensity will reach optimal outcomes when the pH of rare earth upconversion nanoparticles Na YF4:Yb3+/Tm3+ was 4,and the uniform particle size was in 30 nm~50 nm.Under the excitation light at 980 nm,the nano-particle will emit out light in 489 nm.After surface-modified with SiO2,the luminous intensity can be improved further.On the up conversion particles(UCNPs),PEI and HA was connected and named as UCNPs/PEIHA.The cytotoxicity test shows that the toxicity of the probe delivery system to the cells is low when UCNPs/PEI was 1:10 and the cell concentration of UCNPs/PEI particles is 10 g/ml;and the content of PEI is high when N/P was 6:1,miR-26 a is the most complete combination with the carrier using the UCNPs/PEI-HA to transfect mi RNA-26 a into cells.By fluorescence microscope,it was found that the cells glowed green,which show that the GFP gene has been expressed and the vectors has been introduced into cells.The expression of miR-26 a was done by qRT-PCR.It was found that the expression of miR-26 a was significantly up-regulated at 48 h after transfection.The expression of CCND2 and CCNE2 protein was detected by Western blot,and the expression of CCND2 and CCNE2 protein was significantly decreased.The results showed that miRNA-26 a effectively played a role of gene silencing.The nanoparticles emit light delivery systems.In conclusion,this probe delivery system could treat tumor and provide a new approach for tumor therapy. |
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