Cu₂O/MoS₂ Mimic Protease And Its Effect On Human Ovarian Cancer Cells

As a special kind of protein,enzymes play an important role in life activities.However,natural enzymes have many problems,such as difficult extraction and purification,difficult storage,and high production costs.Mimic enzymes with enzyme-like activities have been researched widely as they offer unparalleled advantages in terms of low cost,superior activity,and high stability.Cancer is a disease with a high mortality rate in the world today.The improvement of the survival rate of patients mainly lies in the early detection of cancer and the treatment with new anti-cancer drugs.From the perspective of proteomics,the abnormal expression of related proteins in the human body is often accompanied by the occurrence of related diseases.Utilizing the inherent protease properties of nanoparticles can effectively regulate the microenvironment around cancer cells to achieve the purpose of anti-tumor.This article takes Cu₂O/MoS₂ as the research object,and explores its activity as a mimic protease and its effect on ovarian cancer cells（SKOV3）.（1）Cu₂O/MoS₂ is used as a catalyst to explore its catalytic hydrolysis of bovine serum albumin（BSA）.The effects of catalytic hydrolysis temperature,time,p H,and catalyst concentration have been studied.Analyzing its K_m and k_cat through steady-state kinetic parameters,the analysis data shows that Cu₂O/MoS₂ K_m is lower than trypsin,indicating that its affinity for BSA is higher than trypsin,and the corresponding k_cat is higher than trypsin,indicating that its catalytic performance is also higher then trypsin.Hydrolysis of BSA for 6 times can still maintain more than 70%of the catalytic ability.（2）Cu₂O/MoS₂ was used as a catalyst to catalyze the hydrolysis of casein,and the factors affecting the catalytic hydrolysis were studied.The effect of reaction temperature and reaction time on the hydrolysis of casein was studied.The K_m and k_catvalues are analyzed from the perspective of steady-state dynamics.In the section discussing catalyst stability,it can be seen that it can still maintain more than 70%of the catalytic activity after repeated hydrolysis 5 times.（3）The anticancer activity of Cu₂O/MoS₂ was investigated by in vitro cell experiments.The cytotoxicity of Cu₂O/MoS₂ in five cell lines（human ovarian cancer cells（SKOV3）,human cervical cancer cells（He La）,human lung adenocarcinoma cells（A549）,human colorectal cancer cells（HT-29）,normal mouse embryonic ibroblasts cells（BABL-3T3））.The MTT method was used to explore the Cu₂O/MoS₂ inhibitory effect on cell proliferation.The drugs used to study the inhibitory effect of cell proliferation are paclitaxel（TAX）and carboplatin（DDP）.The AO/EB and Hoechst33342 staining found that the material has the ability to induce apoptosis.Further analysis by flow cytometry showed that the material has the characteristics of inducing apoptosis.Scratch experiments on cancer cells in vitro showed that the number of cell migration decreased significantly with the increase of Cu₂O/MoS₂ concentration and time.Western Blot found that the material has the characteristics of down-regulating matrix metalloproteinase MMP-2/MMP-9.