The lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), the labeled lymphatic vessel density (LVD) and Flt-4-positive vessel density (FVD) were also measured and analyzed relative to the clinicopathological features of the tumors. Our study explored the roles of VEGF-C, VEGF-D, and FLt-4 in the lymphatic metastasis of early-stage cervical cancer. Materials and methods Patients and tissue
samples Patients with cervical carcinoma who were treated between September 2007 and February 2009 were enrolled in this study (n = 97). The tissue samples were obtained at the time of surgery from the Department of Gynecology, Qilu Hospital, Shandong University. Samples and clinical data were collected after informed consent was obtained. Tissues were fixed with
4% paraformaldehyde and paraffin-embedded BKM120 chemical structure for further analysis. The pathological examination verified that no radio- or chemotherapy was received before surgery. Our study was approved by the Ethics Committee of Shandong University. All patients with early-stage invasive cervical cancer were staged according to the 2000 International Federation of Gynecology and Obstetrics (FIGO) staging system. Sixteen of the patients had cervical cancer classified as FIGO stage Ia, 33 as FIGO stage Ib, and 48 as FIGO stage IIa. Based on the analysis of cellular differentiation, 21 cases were HG1, 31 were HG2, and 45 were HG3. Of all the cases, 81 were squamous cell carcinomas
and 16 were adeno-carcinomas. All the patients received pelvic or para-abdominal aortic lymphadenectomy Montelukast Sodium and in total 2376 lymph SN-38 molecular weight nodes were dissected (mean 24.5, median 24.0). A histological review confirmed that 30 cases (30.9%) showed lymph node metastasis and 75 lymph nodes were metastasis positive (mean 2.5, median 2). The age of the patients varied from 26 to 70, with a median value of 42. Of all the patients, 68 were premenopausal and 29 were postmenopausal. The standard for lymphatic vessel invasion was the detection of cancer cells in the cavity of the lymphatic vessel by light microscopy. By this standard, 39 cases showed lymphatic vessel invasion and 58 were negative. All tissue specimens and slides were examined by experienced pathologists. Reagents The reagents used in this study Akt inhibitor in vivo included: rabbit anti-human VEGF-C polyclonal antibody from Zhongshan Goldenbridge Biotech (Beijing, China; catalog no. ZA-0266, 1:50 dilution); rabbit anti-human VEGF-D polyclonal antibody from Boster Inc. (Wuhan, Hubei, China; catalog no. BA1461, 1:100 dilution); rabbit anti-human Flt-4 polyclonal antibody from Abcam (Cambridge, MA, USA; catalog no. ab27278, 1:200 dilution); rabbit anti-human LYVE-1 polyclonal antibody from Abcam (Cambridge, MA, USA; catalog no. ab36993, 1:80 dilution); and an immunohistochemistry SP kit from Jingmei Inc. (Shanghai, China; catalog no. LHK612).