Experimental Study On The Photothermal Effect Of Copper Sulfide Nanoparticls For Cervical Cancer Cell

Objective:To investigate the photothermal killing effect of copper sulfide nanoparticles on the cervical cancer cells and its mechanism.Methods:Copper sulfide nanoparticles(BSA-CuS NPs)were synthesized using bovine serum albumin(BSA)as the templateand characterized by physical method.The cytotoxicity of different concentrations(1.5,3,6,12,24,48 μg/m L)BSA-Cu S NPs was tested by CCK-8 assay.Male Kunming mice were randomly divided into the six groups according to their weight,and the tail vein was injected with different doses of BSA-Cu S NPs(0,6,12,18,24,30 m L/kg).Two weeks,HE staining was used to observe the injury of liver and spleen and kidney.The cell viability of He La cells under near infrared light wavelength of the different power density(0.6,0.8,1.0,1.2,1.4,1.6,1.8,2.0W/cm2)with or without the BSA-Cu S NPs solution was tested by CCK-8 assay.He La cells were divided into control group,Cu S group,NIR group and combined group,the concentration of BSA-Cu S NPs is 24μg/m L and the power density is 1.0W/cm2.Optical microscope,FCM and TUNEL method were used to detect apoptosis.The content of HSP 70 was detected by ELISA;the content of ROS tested by FCM,the contents of SOD and GSH-Px were determined by microenzyme labeling assay;the expression of Bax,Bcl-2,Cleaved Caspase3 and Cytochrome C protein were measured by Western-blot;the level of BRIC5 and TP53were measure by q-PCR.Results:The morphology of the as-synthesized BSA-CuS NPs is lamellar,with an average diameter of 26.25±1.86 nm and a broad band peak at around 980 nm.And all recorded characteristic peaks can be corresponded to the hexagonal phase of standard Cu S.The heating curves illustrated that the temperature of BSA-Cu S NPs rised faster than pure water and in laser intensity-dependent manners.The result of cellular toxicity test indicated that BSA-Cu S NPs were nontoxic to the growth of cells after culture 72 h under 24μg/m L.The healthy adult male Kunming mice did not die after different doses of BSA-Cu S NPs were injected into the tail vein.Two weeks later,HE observation of spleen and kidney did not appear obvious damage.The viability of He La cells treated with BSA-Cu S NPs solution under laser irradiation was significantly lower than laser alone.After different treatments,the combined group was significantly enhanced in apoptosis,and the difference was statistically significant(P<0.05).The content of HSP 70 in the combined group was higer than other groups(P<0.05);The analysis of oxidative stress proteins indicated that compared with the other groups,the combined group had a significant reduction in SOD and GSH-Px activity and increse in ROS level(P<0.05).The result of Western-blot showed that compared with the control group,the Cu S group and the NIR group,the levels of Cytochrome C,Cleaved Caspase3 and Bax/Bcl-2 were up regulated in the combined group,the difference was significant(P<0.05).The result of q-PCR showed that the combined group can attenuates the expression of BIRC 5 gene and improve the TP 53 gene significantly compare with the other group(P<0.05).Conclusion: BSA-CuS NPs combined with near infrared light irradiation has photothermal killing effect on the He La cells.The possible mechanism may be related to free radical and mitochondrial apoptosis pathway.