The clinicopathological significance of ubiquitin-conjugating enzyme E2C, leucine-rich repeated-containing G protein-coupled receptor, WW domain-containing oxidoreductase, and vasculogenic mimicry in invasive breast carcinoma

Abstract

Ubiquitin-conjugating enzyme E2C (UBE2C), a crucial part of the ubiquitin-conjugating enzyme complex, is reported to promote progression of various cancers. Leucine-rich repeated-containing G protein-coupled receptor (LGR5), a biomarker of cancer stem cells, is reported to be responsible for the initiation and progression of cancers. WW domain-containing oxidoreductase (WWOX), a suppressor of tumor, is reported to inhibit initiation and progression of cancers. Vasculogenic mimicry (VM), a new blood supply pattern, is associated with progression of cancers. However, the clinicopathological significance of UBE2C, LGR5, WWOX, and VM in invasive breast carcinoma (IBC) remains elusive. The aim of this study is to investigate the positive rate of UBE2C, LGR5, WWOX, and VM in IBC and their clinical significance.Positive rates of UBE2C, LGR5, WWOX, and VM in 247 whole IBC samples were detected through immunohistochemistry. Patients data (including clinical, demography, follow-up) were collected.Levels of UBE2C, LGR5, VM, and microvessel density (MVD) were significantly higher, and level of WWOX was significantly lower in IBC specimens when compared with normal mammary gland tissues. Levels of UBE2C, LGR5, VM, and MVD were all positively associated with tumor stages, lymph node metastasis (LNM) stages, tumor grades, and tumor-node-metastasis (TNM) stages, and unfavorably with patients' overall survival (OS) and disease-free survival (DFS). Level of WWOX was negatively associated with tumor stages, LNM stages, grades, and TNM stages, and favorably with patients' OS and DFS. Multivariate analysis indicated that levels of UBE2C, LGR5, VM, MVD, and WWOX, as well as TNM stages were independently prognostic factors for OS and DFS in patients with IBC.UBE2C, LGR5, VM, MVD, and WWOX may be considered as promising indicator of IBC prognosis.

Figure 1

Immunostaining of UBE2C, or LGR5, or WWOX, or VM, or MVD in invasive breast carcinoma or the control tissue. A: Negative staining UBE2C in the control tissue (100 magnification, bar = 100 μm); B: Positive staining of UBE2C in the nuclei and cytoplasm of IBC cells (400 magnification, bar = 100 μm); C: Negative staining of LGR5 in the control tissues (100 magnification, bar = 100 μm); D: Positive staining of LGR5 in cytoplasm and membrane of the IBC cells (400 magnification, bar = 100 μm); E: Positive staining of WWOX in the cytoplasm of control tissues (100 magnification, bar = 100 μm); F: Negative staining of WWOX in the IBC cells (100 magnification, bar = 100 μm). G: Positive staining of VM in the IBC tissues (100 magnification; white arrow is VM structure, red arrow is microvessel, bar = 100 μm); H: Positive staining of MVD in the IBC cells (100 magnification, bar = 100 μm). IBC = invasive breast carcinoma, LGR5 = leucine-rich repeated-containing G protein-coupled receptor, MVD = microvessel density, UBE2C = ubiquitin-conjugating enzyme E2C, VM = vasculogenic mimicry.

Figure 2

Kaplan–Meier analysis of overall survival time of patients with invasive breast carcinoma. A: Overall survival of all patients in relation to UBE2C expression (log-rank = 56.737, P < .001); B: OS of all patients in relation to LGR5 (log-rank = 60.951, P < .001); C: OS of all patients in relation to WWOX expression (log-rank = 80.033, P < .001); D: OS of all patients in relation to VM (log-rank = 34.773, P < .001). E: OS of all patients in relation to MVD (log-rank = 22.534, P < .001); F: OS of all patients in relation to ER (log-rank = 18.999, P < .001); G: OS of all patients in relation to PR (log-rank = 11.569, P = .001); H: OS of all patients in relation to HER2 (log-rank = 37.689, P < .001). In A, B, C, D, E, F, G, and H analyses, the green line represents positive staining of factors (MVD score ≥21 is positive) and the blue line represents negative staining factors (MVD score <21 is negative). LGR5 = leucine-rich repeated-containing G protein-coupled receptor, MVD = microvessel density, OS = overall survival, PR = progesterone receptor, UBE2C = ubiquitin-conjugating enzyme E2C, VM = vasculogenic mimicry.

Figure 3

Kaplan–Meier analysis of disease-free survival time of patients with invasive breast carcinoma. A: Disease-free survival of all patients in relation to UBE2C expression (log-rank = 58.314, P < .0001); B: DFS of all patients in relation to LGR5 (log-rank = 59.612, P < .001); C: DFS of all patients in relation to WWOX expression (log-rank = 82.818, P < .001); D: DFS of all patients in relation to VM (log-rank = 36.745, P < .001). E: DFS of all patients in relation to MVD (log-rank = 21.976, P < .001); F: DFS of all patients in relation to ER (log-rank = 20.135, P < .001); G: DFS of all patients in relation to PR (log-rank = 13.735, P < .001); H: DFS of all patients in relation to HER2 (log-rank = 41.686, P < .001). In A, B, C, D, E, F, G, and H analyses, the green line represents positive staining of factors (MVD score ≥21 is positive) and the blue line represents negative staining factors (MVD score <21 is negative). DFS = disease-free survival, HER2 = human epithelial growth factor receptor 2, LGR5 = leucine-rich repeated-containing G protein-coupled receptor, MVD = microvessel density, PR = progesterone receptor, UBE2C = ubiquitin-conjugating enzyme E2C, VM = vasculogenic mimicry.