title: Semaphorin 4D expression is associated with a poor clinical outcome in cervical cancer patients.
abstract: Lymphangiogenesis is thought to be essential for cancer progression, making it an important target in cancer therapy. Lymphangiogenic factors (VEGF-C and VEGF-D) are upregulated in various tumors/cancers, and play an important role in lymphangiogenesis and lymph node metastasis. Similarly, semaphorin 4D (Sema4D) is a potent inducer of angiogenesis, and its overexpression is associated with tumor progression and poor prognosis in a variety of malignancies. However, little is known regarding the functional relationship between Sema4D and VEGF-C/VEGF-D and in the mediation of lymphangiogenesis and lymph node metastasis and clinical outcome. The current study aimed to evaluate the effect of Sema4D expression on outcome in patients with cervical cancer, and to explore the molecular mechanism of Sema4D in tumor progression. We evaluated Sema4D expression, density of lymphatic vessels, and invasion of lymphatic vessels with immunohistochemical methods in 232 human cervical cancers with long-term follow-up. Sema4D expression was correlated with patho-clinical parameters and patients' outcome. A cervical cancer cell line was used to investigate the contribution of sema4D to tumor progression by studying the role of Sema4D in VEGF-C/-D and cell migration using reverse transcription-polymerase chain reaction and Western blotting. We observed that Sema4D expression was higher in metastatic cervical cancer than in nonmetastatic cervical cancer (P<0.001). CD34-positive or D2-40-positive lymphatic vessel density was significantly increased in cases with lymph node metastasis compared with those without lymph node metastasis. The increased Sema4D expression was associated with VEGF-C/-D, the presence of lymphatic invasion, the occurrence of lymph node metastasis, and FIGO stage. We also observed a novel association between Sema4D upregulation and poor prognosis in cervical cancer. In vitro, the Sema4D inhibitory antibody and Sema4D-shRNA suppressed VEGF-C and VEGF-D in the human cervical carcinoma cell lines HeLa, Siha, and Caski cells. Invasiveness assay demonstrated that Sema4D could augment the invasive potential of the tumor cells in the cervical cancer lines and induction of cellular invasiveness by Sema4D stimulation could be inhibited by knockdown of plexinB1 by siRNA. Further mechanistic investigations of tumor cell invasiveness showed that Sema4D could induce activation of GTPase Ras homolog gene family, member A (RhoA), MAPK and AKT. In addition, plexinB1 knockdown by siRNA could suppress the Sema4D signal transmitted to MAPK and Akt. Taken together, these results suggest that Sema4D autocrine within tumor cells contributes to enhanced invasion and tumor progression through increased motility of cervical cancer and VEGF-C/-D-mediated lymphangiogenesis. Sema4D might be useful as a molecular marker of poor prognosis in cervical cancer.
authors.affiliation: Department of Anatomy, Harbin Medical University, 194 Xuefu Road, 150081 Harbin, China.
authors.forename: Huidong
authors.initials: H
authors.lastname: Liu
authors.affiliation: Cancer Institute, Harbin Medical University, 150 Harping Road, 150081 Harbin, China.
authors.forename: Yanmei
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Department of Anatomy, Harbin Medical University, 194 Xuefu Road, 150081 Harbin, China.
authors.forename: Jianbing
authors.initials: J
authors.lastname: Xiao
authors.affiliation: Department of Anatomy, Harbin Medical University, 194 Xuefu Road, 150081 Harbin, China.
authors.forename: Shucai
authors.initials: S
authors.lastname: Yang
authors.affiliation: Department of General Surgery, Mudanjiang First Hospital, 4 Jianwei Road, 157011 Mudanjiang, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Radiotherapy, The Third Affiliated Hospital of Harbin Medical University, 150 Harping Road, 150081 Harbin, China.
authors.forename: Wenhui
authors.initials: W
authors.lastname: Kang
authors.affiliation: Department of Anatomy, Harbin Medical University, 194 Xuefu Road, 150081 Harbin, China.
authors.forename: Xinlei
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Medical Microbiology, Heilongjiang Province Key Laboratory for Immunity and Infection, Harbin Medical University, 194 Xuefu Road, 150081 Harbin, China. Electronic address: fengminzhang2013@163.com.
authors.forename: Fengmin
authors.initials: F
authors.lastname: Zhang
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000378:metabolism* / Q000556:secondary; D000328:Adult; D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D018196:Carcinoma, Adenosquamous / Q000235:genetics / Q000378:metabolism* / Q000556:secondary; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism* / Q000556:secondary; D002465:Cell Movement; D005260:Female; D006367:HeLa Cells; D006801:Humans; D053208:Kaplan-Meier Estimate; D042583:Lymphangiogenesis; D008207:Lymphatic Metastasis; D042601:Lymphatic Vessels / Q000378:metabolism* / Q000473:pathology; D008875:Middle Aged; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D009361:Neoplasm Invasiveness; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D011379:Prognosis; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D034622:RNA Interference; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D039961:Semaphorins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection; D015854:Up-Regulation; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D042582:Vascular Endothelial Growth Factor C / Q000378:metabolism; D042643:Vascular Endothelial Growth Factor D / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D014408:Biomarkers, Tumor; C093224:CD100 antigen; D009419:Nerve Tissue Proteins; C492119:PLXNB1 protein, human; D011956:Receptors, Cell Surface; D039961:Semaphorins; C467486:VEGFC protein, human; C579459:VEGFD protein, human; D042582:Vascular Endothelial Growth Factor C; D042643:Vascular Endothelial Growth Factor D; C066737:RHOA protein, human; D051057:Proto-Oncogene Proteins c-akt; D020928:Mitogen-Activated Protein Kinases; D020742:rhoA GTP-Binding Protein
keywords: Cervical cancer; Invasiveness; Lymphangiogenesis; Outcome; Semaphorin 4D; VEGF-C; VEGF-D