and bone tissues are the most frequent sites for metastases. Up to 70% of patients with metastatic breast cancer develop bone metastases that induce increased osteoclast activity, resulting in local bone destruction and skeletal complications, including pain, hypercalcemia, and nerve compression (1-3). The development and outgrowth of these secondary lesions rely upon the Wei Yan et al. www.boneresearch.org | Bone Research 363intricate cellular and molecular interactions between mammary tumor cells in the bone microenvironment. Tumor cells secrete signaling proteins, such as parathyroid hormone-related peptide (PTHrP) (4), to promote the secretion of RANKL, the receptor activator of nuclear factor-κB ligand, in osteoblast cells that activates bone osteolysis induced by osteoclasts. The concomitant bone destruction releases various growth factors, including transforming growth factor-β (TGF-β) and insulin-like growth factor-1 (IGF-1), to further bind the receptors on the surface of tumor cells and enhance cell proliferation and PTHrP production through activation of the Smad/ MAPK signaling transduction pathways, thus establishing a vicious cycle of bone metastases (5). The most widely used treatments in skeletal complications of varied neoplastic diseases are bisphosphonates (BPs) that block osteoclast activity; this application has been effective in decreasing formation of bone lesions, although this strategy fails to induce the bone regeneration (6). Furthermore, a growing number of case reports have demonstrated that long-term BP therapy might lead to osteonecrosis of the jaw (ONJ) (7-10). Therefore, the search for treatment options that can effectively inhibit tumor growth and reduce bone destruction caused by tumor metastases with mitigated side effects is of pivotal significance.Plumbagin (5-hydroxy-2-methyl-1,4-naphthoquinone), derived from the roots of the medical plant plumbago zeylanica, is one of the most investigated compounds. Recent studies have reported the antibacterial effects (11), anti-inflammatory effects (12), and anticancer activities of plumbagin both in vitro (13) and in vivo (14)(15). Previous studies have shown that plumbagin can inhibit tumor cell proliferation in vitro by inducing apoptosis and autophagy of breast neoplasm cells (13) and the invasion of prostate cancer cells (14). Recent studies have also demonstrated that plumbagin can inhibits osteoclastogenesis and reduces human breast cancerinduced osteolytic bone metastasis (16-18), however, more in vivo evidence need to be supplemented and the relevant mechanisms still remain to be investigated. Therefore, the aim of this study is to investigate whether there are any suppressive effects of plumbagin on the invasion and migration of human breast cancer MDA-MB-231SArfp cells and to explore potential functional mechanisms. We thus established an animal model of breast cancer bone metastases via injection of breast cancer cells intracardially. A non-invasive small animal monitoring system in addition to X-ray imaging and histologica...

Galectin-3 (Gal-3) has been found to be involved in the tumor progression and chemoresistance of epithelial ovarian cancer (EOC). Some studies have shown that Gal-3 may interact with nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB). However, it is unclear whether the effects of Gal-3 on the metastasis and chemosensitivity of EOC are related to NF-κB. In this study, we aimed to explore whether Gal-3 promoted progression and carboplatin resistance in EOC via NF-κB pathway. Plasmid transfection and RNA interference were used to upregulate or downregulate the expression of Gal-3 in ovarian cancer cell lines. Then, the expression of Gal-3 and the protein expressions of phosphorylation NF-κB pathway molecules were further detected by Western blot. Transwell migration assay was employed to detect the effects of Gal-3 on the migration and invasion of ovarian cancer cell lines. After treatment with carboplatin, flow cytometry (FCM) was employed to detect the effects of Gal-3 on carboplatin-induced apoptosis. Immunofluorescence technique was used to examine the translocation of phosphorylated P65 into the nucleus in ovarian cancer cells after the upregulation of Gal-3. After the knockdown of Gal-3 by small interfering RNA (siRNA), the migration and the invasion of cancer cells were significantly inhibited while the apoptosis and the sensitivities to carboplatin increased. Western blot showed reduction in the phosphorylation components of the NF-κB pathway: inhibitor of kappa B (IκB), IκB kinase (IKK), and P65. However, after the Gal-3 upregulation by plasmid transfection, the capabilities of migration and invasion of cancer cells were significantly promoted while the apoptosis and the sensitivities to carboplatin decreased. Immunofluorescence showed increased nuclear translocation of P65. Inhibitors of the NF-κB pathway did not affect the Gal-3 expression level in ovarian cancer cells. Gal-3 may affect the migratory and invasive capabilities of cancer cells as well as the chemosensitiviy to carboplatin in EOC by acting through the NF-κB pathway.

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Purpose The role of HHLA2, a new immune checkpoint ligand, is gradually being elucidated in various solid tumours. However, its role in ovarian cancer remains unclear; thus, its expression profile and clinical significance in ovarian cancer must be examined. Methods We performed immunohistochemistry to examine HHLA2 expression in 64 ovarian cancer tissues and 16 normal ovarian tissues. The relationships between HHLA2 expression and clinicopathological features, prognosis, and CD8+ tumour-infiltrating lymphocytes (TILs) in patients were analysed. Additionally, the Cancer Cell Line Encyclopedia database was used to analyse the correlation between HHLA2 expression and PD-L1 or B7x expression. Furthermore, the biological function of HHLA2 in ovarian cancer cells was initially explored. Results Only 17.2% of ovarian cancer patients showed HHLA2 expression, which was significantly associated with the differentiation of ovarian cancer cells (p = 0.027), and well-differentiated tumours expressed higher levels of HHLA2. The density of CD8+ TIL was associated with increased HHLA2 expression (p = 0.017), and the CD8+ TIL count was higher in the HHLA2-positive group than that in the HHLA2-negative group (p = 0.023). Moreover, multivariate analysis identified HHLA2 expression as an independent prognostic factor that predicted improved survival (p = 0.049; HR = 0.156; 95% CI = 0.025–0.992). Additionally, we also found that overexpressing HHLA2 inhibited the proliferation of ovarian cancer cells. Conclusion HHLA2 is associated with tumour differentiation and high CD8+ TIL levels; and predicts improved survival in ovarian cancer. Along with previously reported findings that HHLA2 behaves as a co-stimulatory ligand, our study suggests that the loss of HHLA2 may contribute to the immunosuppressive microenvironment and progression of ovarian cancer.

Aberrant activation of the canonical Wnt pathway plays a significant role in cervical cancer (CC). However, limited data show the correlation between the cancer clinicopathological characteristics and the key molecules such as β-catenin and Wnt inhibitory factor 1 (WIF1). In this study, β-catenin and WIF1 expression were analyzed by immunohistochemistry for 196 patients with CC, 39 with cervical intraepithelial neoplasia (CIN), and 41 with normal cervical epithelium (NCE). Significant overexpression of β-catenin was detected in CC (67.9%) when compared to CIN (43.6%) or NCE (34.1%), p < 0.01, while low WIF1 expression was detected in CC (24.0%) when compared to CIN (59.0%) or NCE (58.5%), p < 0.001. Negative correlation was shown between β-catenin and WIF1 expression (r = −0.637, p < 0.001). In addition, multivariate analysis revealed that both lymph node metastasis and β-catenin expression were the independent prognostic factors not only for disease-free survival (HR = 5.029, p < 0.001; HR = 2.588, p = 0.035, resp.), but also for overall survival (HR = 5.058, p < 0.001; HR = 2.873, p = 0.031, resp.). Our findings indicate that, besides lymph node metastasis, β-catenin expression may also be a poor prognostic factor for CC while WIF1 could be a potential drug target for treatment of advanced CC.

PurposeThis study was designed to explore the expression levels of Galectin-3 (Gal-3) and β-catenin in serous epithelial ovarian cancer (SEOC), the linkage between their expressions, and the clinicopathological features of SEOC patients.Patients and methodsSeventy-four SEOC patients’ specimens were detected for Gal-3 and β-Catenin expressions using immunohistochemistry, and the association between β-catenin or Gal-3 protein expressions and clinicopathological features, treatment effects, and prognosis were analyzed using SPSS 19.0. Western blot was used to analyze protein expressions of Wnt/β-catenin pathway in ovarian cancer cell lines.ResultsThere was a statistically significant positive correlation between Gal-3 and β-catenin expressions in SEOC (r=0.304 and P=0.001). Gal-3 expression was related to the grade (P=0.037), clinical stage (P=0.034), platinum resistance (P=0.030), and recurrence (P=0.001) in SEOC. There was a significant correlation between β-catenin with recurrence in SEOC (P=0.035). Platinum resistance (P=0.003) and Gal-3 expression (P<0.001) were independent risk factors for poorer overall survival (OS). OS of the strongly positive Gal-3 group was significantly lower than that of the negative and weakly positive groups (log-rank test, P=0.001). OS of the positive β-catenin group was lower than that of the negative β-catenin group (log-rank test, P=0.034). Downregulating Gal-3 expression attenuated the protein expressions of Wnt/β-catenin pathway in ovarian cancer cell lines.ConclusionGal-3 might activate Wnt/β-catenin signaling pathway in SEOC. Hence, Gal-3 may serve as a prognostic factor for SEOC. Targeting Gal-3 may be a promising new treatment approach for SEOC.

Objective Evidence on uterine serous cancer (USC) prognosis has been limited and inconclusive. We aim to explore the survival benefits of comprehensive lymphadenectomy in USC patients after surgery and develop a prognostic nomogram to predict survival. Materials and Methods USC patients who had undergone hysterectomy between 2010 and 2015 were identified from Surveillance, Epidemiology and End Results (SEER) database. The relationship between the extent of lymphadenectomy and survival, including overall survival (OS) and cancerspecific survival (CSS), was estimated with Kaplan-Meier (K-M) analysis. Univariate and multivariate Cox regression analyses were utilized to determine the independent prognostic factors. A nomogram was then developed, calibrated and internally validated. Results A total of 2853 patients were identified. K-M survival analysis revealed that patients with ≥12 pelvic lymph nodes (PLNs) removed had significantly better OS and CSS than those without (both P <0.001). However, patients with ≥6 para-aortic lymph nodes removed was not associated with similar survival benefits than patients without (P >0.1). Multivariate analyses for OS and CSS revealed that age, T stage, N stage, tumor size, adjuvant therapy and ≥12 PLNs removed were independent prognostic factors (all P<0.05) and were subsequently incorporated into the nomogram. The Harrell's C-index of the nomogram was significantly higher than that of the International Federation of Gynecology and Obstetrics staging system (OS: 0.739 vs 0.671, P <0.001; CSS: 0.752 vs 0.695, P <0.001). Furthermore, the nomogram was well calibrated with satisfactory consistency. Conclusions Comprehensive pelvic lymphadenectomy should be recommended to USC patients for its survival benefits. And a nomogram has been developed to predict the survivals of USC patients after surgery.

BackgroundOccult invasive cervical cancer discovered after simple hysterectomy is not common, radical parametrectomy (RP) is a preferred option for young women. However, the morbidity of RP was high. The aim of our study is to assess the incidence of parametrial involvement in patients who underwent radical parametrectomy for occult cervical cancer or radical hysterectomy for early-stage cervical cancer and to suggest an algorithm for the triage of patients with occult cervical cancer to avoid RP.MethodsA total of 13 patients with occult cervical cancer who had undergone RP with an upper vaginectomy and pelvic lymphadenectomy were included in this retrospective study. Data on the clinicopathologic characteristics of the cases were collected. The published literature was also reviewed, and low risk factors for parametrial involvement in early-stage cervical cancer were analyzed.ResultsOf the 13 patients, 9 had a stage IB1 lesion, and 4 had a stage IA2 lesion. There were four patients with grade 1 disease, seven with grade 2 disease, and two with grade 3 disease. The median age of the entire patients was 41 years. The most common indication for extrafascial hysterectomy was cervical intraepithelial neoplasia 3. Three patients had visible lesions measuring 10–30 mm, in diameter and ten patients had cervical stromal invasions with depths ranging from 4 to 9 mm; only one patient had more than 50% stromal invasion, and four patients had lymph-vascular space invasion (LVSI). Perioperative complications included intraoperative bowel injury, blood transfusion, vesico-vaginal fistula, and ileus (1 case for each). Postoperative pathologic examination results did not show residual disease or parametrial involvement. One patient with positive lymph nodes received concurrent radiation therapy. Only one patient experienced recurrence.ConclusionsPerioperative complications following RP were common, whereas the incidence of parametrial involvement was very low among selected early-stage cervical cancer patients. Based on these results, we thought that patients with very low-risk parametrial involvement(tumor size ≤2 cm, no LVSI, less than 50% stromal invasion, negative lymph nodes) may benefit from omitting RP. Further prospective data are warranted.