The Effect Of Dexmedetomidine On Proliferation And Migration Of Hepatocellular Carcinoma Cells

Background and Objective:Hepatocellular carcinoma(HCC)is one of the most common malignant tumors in the world.It is concentrated in Asia and Africa.The incidence of HCC is increasing sharply year by year worldwide.Hepatocellular carcinoma has a very poor prognosis,with a5-year survival rate much lower than that of other malignancies.Although with the development of medical technology and the improvement of the diagnosis and treatment of liver cancer,surgical resection is still the most effective treatment for liver cancer patients.Most of the patients died of metastasis and recurrence of tumor after hepatocellular carcinoma surgery.Therefore,it is of great significance to explore the influencing factors of liver cell metastasis and recurrence.A large number of studies have shown that the application of perioperative anesthetic drugs and strategies can directly affect the proliferation and immunity of cells,thus indirectly interfering with the metastasis and recurrence of tumor cells.The application of general anesthesia plays an important role in the long-term outcome of cancer patients.Therefore,it is of great clinical significance to explore the interaction between anesthesia and tumor and its molecular mechanism.Dexmedetomidine,a highly selective alpha2 adrenergic receptor agonist,has been widely used in clinical practice as an adjunct to anesthesia in recent years.It has been found that dexmedetomidine can promote the proliferation and invasion of lung cancer and glioma cells.However,the interaction between dexmedetomidine and liver cancer has rarely been reported.SIRT1 is a highly conserved nicotinamide adenine dinucleotide dependent histone deacetylase.It plays an important role in the regulation of cellular senescence,metabolism and immune response.However,the role of SIRT1 in tumor is still very controversial.Its ability to promote and inhibit tumor is specific for cells and organs.Studies have shown that dexmedetomidine can maintain cardiac homeostasis through SIRT1,and whether dexmedetomidine can maintain and promote the function of HCC cells through SIRT1 has not been reported yet.Apoptosis is an orderly and autonomous cell death regulated by genes.The Bcl-2 family plays a key role in apoptosis.It has been reported that SIRT1 can regulate the growth and development of cells through apoptosis signaling pathway.In this study,we studied the changes of proliferation and migration of HCC Huh-7 cells,the changes of intracellular SIRT1 expression and apoptosis expression under the effect of dexmedetomidine,and explored the effect of SIRT1-mediated apoptosis inhibition on the proliferation and migration of HCC Huh-7 cells.Methods:The experiment was divided into two parts: cells and animals.First,Huh-7 cells from HCC in logarithmic growth phase were selected for the following studies:1.The Huh-7 cells were treated with different concentrations of dexmedetomidine for24 h,and the proliferation activity of Huh-7 cells was detected by MTT assay.Transwell method was used to detect the migration ability of Huh-7 cells.Western Blot was used to detect the expression of SIRT1 and Bcl-2/Bax in Huh-7.2.Si-SIRT1 was transfected into Huh-7 cells,and the transfection efficiency was detected by QT-PCR 6 h later.After confirming that the transfection efficiency reaches70%,proceed to the next step.Dexmedetomidine was added for treatment for 24 h.MTT assay was used to detect the proliferation activity of Huh-7.Transwell method was used to detect the migration ability of Huh-7 cells.Western Blot was used to detect the expression of Bcl-2/Bax in Huh-7.Secondly,6 BALB/c-nu male SPF mice were randomly divided into control group and dexmedetomidine group.Four days after subcutaneous implantation of Huh-7 cells,the cells were injected with 0.5 mg/kg dexmedetomidine or corresponding volume of normal saline.The injection was given once every two days for 7 consecutive times.The nude mice were sacrificed 44 days after cell implantation,and the body weight,mass and volume of subcutaneous tumor of the nude mice were observed Results:1.Dexmedetomidine can enhance the proliferation and migration of Huh-7 cells.Dexmedetomidine increased the activity level of Huh-7 cells,and reached the maximum effect at the concentration of 0.1 n M,so 0.1 n M was selected as the optimal effective concentration.0.1 n M dexmedetomidine induced increased migration of HCC Huh-7cells and increased intracellular SIRT1 expression and the expression of Bcl-2/Bax.2.Downregulation of intracellular SIRT1 level can reduce the effect of dexmedetomidine on promoting proliferation and migration and the expression of Bcl-2/Bax.Dexmedetomidine showed no significant changes in cell proliferation and migration and the expression of Bcl-2/Bax after knockdown of Huh-7 intracellular level.3.In vivo experiments,the body weight,tumor mass and volume of nude mice treated with dexmedetomidine showed no significant changes.In the tumor formation experiment of nude mice,the animal weight and the body weight of the subcutaneous tumor of nude mice were not significantly changed by intraperitoneal injection of dexmedetomidine.Conclusion:Dexmedetomidine can promote the proliferation and migration of Huh-7 cells.Inhibition of apoptosis is a key factor in the effect of dexmedetomidine on the proliferation and migration of Huh-7 cells.The up-regulation of intracellular SIRT1 expression plays an important role in the promotion of proliferation and migration of Huh-7 cells by dexmedetomidine.