Effects of Anesthetic Methods on Hepatic Cancer Cell Function in Vitro

Effects of Serum From Hepatocellular Carcinoma Surgery Patients Under Different Anesthetic Methods on Hepatic Cancer Cell Function in Vitro

Numerous studies find that anesthetic methods may influence the recurrence of tumor and the overall survival of patients after primary cancer surgery. Radiofrequency (RF) ablation is now widely used in the clinic for treatment of hepatocellular carcinoma (HCC). Currently, diverse anesthetic methods, including general anesthesia (GA), epidural anesthesia and local anesthesia (LA), are used for RF ablation surgery. Using serum from HCC surgery patients randomized to receive either GA or LA during surgery, we will investigate the effects of anesthetic methods on cellular invasion, migration and proliferation of HepG2 hepatic cancer cells in vitro. The expression levels of inflammatory cytokines in the serum from patients of both groups will also be analyzed.

Patients diagnosed with hepatocellular carcinoma (HCC) undergoing elective radiofrequency (RF) ablation surgery will be randomly allocated to general anesthesia(GA) group or local anesthesia(LA) group. Patients from GA group will receive anesthesia induced with 0.05-0.1mg/kg intravenous midazolam, 3-6ug/kg fentanyl, 1.0-2.5mg/kg propofol and 0.1-0.2mg/kg atracurium. Laryngeal mask will be used to maintain ventilation. Anesthesia will be maintained with 4-8mg/kg/h propofol and 0.1-0.3ug/kg/min remifentanil, and additional non-depolarizing muscle relaxant when necessary. In patients from LA group, analgesia will be provided by a subcutaneous injection of 2% lidocaine by the surgeons before insertion of the needles. Serum will be collected from patients of both groups at before induction and 1h post-surgery. The HepG2 hepatic cancer cell line will be treated with patient serum from both groups. The effects of anesthetic methods on cellular invasion, migration, proliferation will be measured. The expression levels of inflammatory cytokines in the serum from patients of both groups will also be analyzed.

mean percentage change from post- to pre-operative values of invasion of HepG2 cells cultured in patients' serum

HepG2 cells will be cultured with serum from patients undergoing RF ablation under GA or LA for 24h. Cell invasion will be measured with Matrigel methods. Mean percentage change from post- to pre-operative values of invasion for each individual patient will be calculated and compared between the GA and LA groups.

mean percentage change from post- to pre-operative values of migration of HepG2 cells cultured in patients' serum

HepG2 cells will be cultured with serum from patients undergoing RF ablation under GA or LA for 24h. Cell migration will be measured with Transwell methods. Mean percentage change from post- to pre-operative values of migration for each individual patient will be calculated and compared between the GA and LA groups.

mean percentage change from post- to pre-operative values of proliferation of HepG2 cells cultured in patients' serum

HepG2 cells will be cultured with serum from patients undergoing RF ablation under GA or LA for 24h. Cell proliferation will be measured with EdU methods. Mean percentage change from post- to pre-operative values of proliferation for each individual patient will be calculated and compared between the GA and LA groups.

Inclusion Criteria: 18 to 65 years old ASA grade I-III Diagnosed with a single primary liver tumor of 3cm or smaller Is scheduled for elective radiofrequency ablation surgery With no macro-vascular invasion, no lymph node or extrahepatic metastases Child-Pugh Class A or B Exclusion Criteria: a history of liver surgery previously (including radiofrequency ablation) severe systemic disease (heart, lung, kidney, or immune system) INR>1.5 or platelet count <45,000 cells/mm3 a history of addiction to opioids; Disagree to participate the trial