Precise determining lymphovascular infiltration and the role of tumor-associated macrophage in cervical cancer

Abstract Background: Accurate determination of lymphovascular infiltration(LVI) status can guide the accurate surgical scope of cervical cancer patients and improve the prognosis. But LVI reduces overall survival (OS) of cervical cancer patients and not easy detected by routine HE staining.The role of tumor-associated macrophages (TAMs) in this process is not well defined. Methods: Early stage cervical cancer patients received carbon nanoparticles(CNP) for sentinel lymph nodes(SLNs) mapping,laparotomy pelvic lymph node dissection and radical hysterectomy.The samples were detected for ultra staging,cocktail double immunohistochemical(IHC) staining, Flow cytometry and Western blot anlysis. Results: The combine of CNP mapping,ultra staging and double IHC staining enhance determining ratio for tumor LVI than that of HE staining alone(41.8% (41/98) vs. 20.4% (20/98), P=0.046)). When the number of vascular invasion foci>2.0,or the number of cancer emboli cells ≥5.3, there was negative correlation with the OS for patients (p < 0.05). More M2 macrophage emerged surrounding the tumor vasculature detected by double IHC staining, accompanied higher M2/M1 ratio detected by flow cytometry detection (P<0.05). M2 TAMs number was positive correlation to the number of tumor LVI(P=0.0024),indicating M2 TAMs increased in tumor LVI infiltration.TAMs related genes MMP2,SPARC and GNLY expessed higher level in tumor in Western blot analysis, while the OS of the patients decreased accordingly. Conclusions: The CNP mapping combined with double IHC staining could improve the precision surgical detection rate of cervical cancer LVI. M2 macrophages polarization and related markers increased cervical cancer tissure LVI.