The Inhibiting Effects Of Propofol On Colorectal Cancer Progression Through NF-κB/HIF-1α Signaling Pathway:A Patient Derived Xenograft Based Study

Background and Purpose: Colorectal cancer(CRC)is a neoplastic disease that gradually accumulates and develops through genetic variations and epigenetic changes.Surgical excision is the corner-stone method for CRC therapy.In clinical practice,there are two commonly used anesthesia techniques,namely total intravenous anesthesia or inhalation anesthesia,during surgery.There is accumulating evidence has shown that the two anesthesia methods have different effects on tumor recurrence,metastasis and prognosis after surgery in CRC patients,and furthermore total intravenous anesthesia seems confer beneficial effects,of which decreasing the possibility of the recurrence and metastasis of tumor.Propofol is one of the most frequently used intravenous anesthetics in practice.It remains unknown whether it can reduce the recurrence and metastasis after surgery in cancer patients.Hypoxia inducible factor-1α(HIF-1α)is an important transcription factor involved in the development and progression of various diseases,which is regulated by multiple signaling pathways including nuclear factor-κB(NF-κB)pathway.The overactivation of NF-κB pathway is a significant characteristic of CRC,and phosphorylation of NF-κB dimer enters into nucleus and promotes the transcription of HIF-1α.The present study,using the patient-derived tumor xenograft model(PDX),was to investigate whether propofol could inhibit NF-κB/HIF-1α signaling pathway in vivo and in vitro,and the potential mechanism involving this process,thereby delineating the role of propofol in inhibiting the occurrence and progression of CRC after surgery.Methods: A PDX mouse model incubating CRC cell was established and then sub-cultured to the F3 generation.HE staining and immunohistochemical staining were used to determine the morphological and genetic stability of the PDX model.PDX mouse model F3 was used for intraperitoneal injection of propofol to explore the antitumor effect of propofol in vivo.CRC cell lines(HCT116 and SW480)were partially selected in vitro,and different concentrations of propofol were added into cell culture medium.The proliferation effect of propofol on CRC cell lines was detected by CCK-8 assay.The effect of propofol on migration and invasion of CRC cells was evaluated by scratch healing experiment and Transwell experiment.The inhibitory effects of propofol on NF-κB and HIF-1α in CRC cell lines were determined by Western Blotting and immunofluorescence assay,to further clarify the regulatory effects of propofol on NF-κB and HIF-1α.Results:(1)Model part: The P-F3 generation tumor tissues of CRC PDX model are poorly differentiated intestinal cancer,and the cell morphology,karyotype and density have no obvious changes.Ki67,NF-κB and HIF-1α were all highly expressed(>80%),indicating that histomorphology and molecular phenotype were expressed stably in the early passage.(2)In vivo: Propofol did not significantly inhibit tumor growth in vivo(P=0.7198).However,compared with the blank control group,the expression of NF-κB and HIF-1α was decreased in the propofol treatment group,and there were no significant changes in cell morphology,karyotype and Ki67 expression.(3)In vitro: Propofol significantly inhibited the ability of proliferation,migration and invasion in CRC cell lines(HCT116 and SW480),and there was statistically significance when compared to the control group(P < 0.0001);The expression of NF-κB and HIF-1α decreased gradually with the increase of propofol concentration in both cell lines.After activation and inhibition of NF-κB,the expression of HIF-1αchanged.Further studies showed that propofol inhibited LPS-activated NF-κB expression of HIF-1α was similar to NF-κB inhibitor Bay117082(P < 0.0001).Conclusion: It was revealed that propofol showed no significant tumor suppressive effect in the context of intraperitoneal injection.In vitro,the inhibited effect of propofol on the ability of proliferation,migration and invasion of CRC cells(HCT116 and SW480)was in a dose-dependent manner,possibly by participating in the regulation of NF-κB/HIF-1α signaling pathway.