Effects of Serum From Radiofrequency Ablation Patients Receiving General Anesthesia or Local Anesthesia on Hepatocellular Carcinoma Cancer Cell Malignancy: A Prospective Randomized Controlled Trial

Abstract

Background: Whether anesthesia methods affect malignant biological behavior of cancer remains unresolved. In this study, we aim to compare the effects of general anesthesia (GA) and local anesthesia (LA) on serum collected from primary hepatocellular carcinoma (HCC) patients presenting for radiofrequency ablation (RFA).

Methods: From August 2020 to December 2020, a prospective, randomized, and controlled study was conducted at Renji Hospital, which is affiliated with Shanghai Jiaotong University School of Medicine. 25 qualified patients from 18 to 65 years of age undergoing RFA were enrolled in the study and randomly assigned into two groups: the GA group (n = 14) and the LA group (n = 11). Venous blood was drawn from all patients preoperatively and 1 hour postoperatively. The serum collected was then used for the culturing of HepG2 cells. The malignant biological behaviors of HepG2 cells, including invasion, migration and proliferation, were observed after 24 hours of exposure to patients' serum. ELISA was used to compare expression levels of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α) and lymphokines (IFN-γ, IL-2) in patients' serum from both groups.

Results: HepG2 cells cultured with postoperative serum obtained from patients who received GA, but not LA, were associated with significantly increased cell invasion, migration and proliferation, compared to preoperative serum from the same patient group. Expression levels of pro-inflammatory cytokines were significantly higher, and lymphokines significantly lower in postoperative serum from GA patients compared to the corresponding preoperative serum.

Conclusion: GA affects the serum milieu of patients with HCC, promoting the malignant biological behavior of a human HCC cell line.

Keywords: cancer cell malignancy; general anesthesia; hepatocellular carcinoma; local anesthesia; serum milieu.

Figure 1

Clinical trial flow diagram. 28 patients were recruited and randomized to the LA group or GA group. 25 patients were included in the final analysis, n = 11, LA group, and n = 14, GA group.

Figure 2

Serum from RFA patients receiving GA facilitated cell invasion of HepG2 cells. (A) Representative image of pre-LA serum-treated HepG2 cells; (B) Representative image of pre-GA serum-treated HepG2 cells; (C) Representative image of post-LA serum-treated HepG2 cells; (D) Representative image of post-GA serum-treated HepG2 cells; Original magnification, 100X; (E) Graphical representation of cells that invaded the lower surface of the four groups; (F) Graphical representation of the mean percentage change from post- to preoperative values of invading cell numbers in the GA group vs the LA group. Values are expressed as mean ± SD. n = 11, LA group. n = 14, GA group. **P < 0.01. GA, General Anesthesia; LA, Local Anesthesia.

Figure 3

Serum from RFA patients receiving GA facilitated cell migration of HepG2 cells. (A) Representative image of HepG2 cells cultured with preoperative or 1h postoperative serum from the LA group; (B) Representative image of HepG2 cells cultured with preoperative or 1h postoperative serum from the GA group; (C) Graphical representation of recovery ratios of the four groups in (A, B, D) Graphical representation of the mean percentage change from post- to preoperative values of recovery ratios in the GA group vs the LA group. Values are expressed as mean ± SD. n =11, LA group. n = 14, GA group. **P < 0.01. GA, General Anesthesia; LA, Local Anesthesia.

Figure 4

Serum from RFA patients receiving GA facilitated cell proliferation of HepG2 cells after long-term, but not short-term, exposure. (A) CCK-8 assay OD values (culture time = 24h); (B) CCK-8 assay OD values (culture time = 48h); (C) mean percentage change from post- to preoperative OD value (culture time = 24h); (D) mean percentage change from post- to preoperative OD value (culture time = 48h). Values are expressed as mean ± SD. n = 11, LA group. n = 13~14, GA group. **P < 0.01. GA, General Anesthesia; LA, Local Anesthesia.

Figure 5

Serum from RFA patients receiving GA facilitated cell proliferation of HepG2 cells after long-term exposure, demonstrated with EdU assays. (A) HepG2 cells cultured with preoperative and 1h postoperative serum from the LA and GA groups, respectively; proliferative cells were stained with EdU (red), and nuclei were counterstained with Hoechst 33342 (blue); Original magnification, 100X; (B) Graphical representation of the proportions of EdU positive cells of the four groups; (C) Graphical representation of the mean percentage change from post- to preoperative values in EdU positive cells in the GA vs LA groups. Values are expressed as mean ± SD. n = 11, LA group. n = 14, GA group. **P < 0.01. GA, General Anesthesia; LA, Local Anesthesia.