S100A6 promotes cell proliferation in human nasopharyngeal carcinoma via the p38/MAPK signaling pathway.

An elevated level of S100A6 is associated with poor outcomes of many tumor types, but, how S100A6 contributes to nasopharyngeal carcinoma (NPC) progression remains unknown. Here, we investigated the expression and prognostic significance of S100A6 in NPC and explored the molecular mechanisms under-lying the role of S100A6 in NPC development. The results showed that S100A6 was markedly up-regulated in NPC tissues and cell lines compared to paired peritumoral normal tissues and a normal nasopharyngeal epithelial cell line, respectively. In tissues from 92 NPC patients, high S100A6 expression was associated with advanced N stage, locoregional failure and disease progression and was predictive of poor locoregional recurrence-free survival (LRRFS, P = 0.001) and progression-free survival (PFS, P = 0.001). Multivariate analysis showed that S100A6 is an independent prognostic factor for LRRFS and PFS. Silencing S100A6 using siRNA or shRNA significantly suppressed NPC cell proliferation, colony formation and p38/mitogen-activated protein kinase (MAPK) activity in vitro and inhibited tumor growth in a xenograft mouse model of NPC. In contrast, overexpressing S100A6 via plasmid transfection resulted in increased NPC cell proliferation and p38/MAPK activation. S100A6-induced proliferation was abolished by a p38 inhibitor. In summary, S100A6 may be a new prognostic marker of NPC and may promote NPC development via the activation of p38/MAPK signaling pathways. These findings suggest S100A6/p38/MAPK signaling as a potential therapeutic target for NPC. © 2016 Wiley Periodicals, Inc.

Risk factors and prediction-score model for distant metastasis in nasopharyngeal carcinoma treated with intensity-modulated radiotherapy.

The objective of this study is to identify the risk factors and construct a prediction-score model for distant metastasis (DM) in nasopharyngeal carcinoma (NPC) patients treated with intensity-modulated radiotherapy (IMRT). A total of 520 nonmetastatic NPC patients were analysed retrospectively. The independent risk factors for DM were tested by multivariate Cox regression analysis. The prediction-score model was established according to the regression coefficient. The median follow-up was 88.4 months. The 5-year DM rate was 15.1%. N2-3, primary tumour volume of nasopharynx (GTVnx) >24.56 cm(3), haemoglobin change after treatment (ΔHGB) >25.8 g/L, albumin-globulin ratio (AGR) ≤1.34, pretreatment neutrophil-lymphocyte ratio (NLR) >2.81 and pretreatment serum lactate dehydrogenase (LDH) >245 U/L were significantly adverse independent predictive factors for DM. Three subgroups were defined based on the prediction-score model: low risk (0-2), intermediate risk (3-4) and high risk (5-8). The 5-year DM rates were 4.6, 21.8 and 50.8%, respectively (P < 0.001). The areas under the curve for DM in the prediction-score model and the UICC/AJCC staging system seventh edition were 0.748 and 0.627, respectively (P < 0.001). The scoring model is useful in evaluating the risk of DM in IMRT-treated NPC patients and guiding future therapeutic trials. Further prospective study is needed.

MicroRNA-1269 promotes proliferation in human hepatocellular carcinoma via downregulation of FOXO1.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignancies and a major cause of cancer-related mortality in the world. MicroRNAs (miRNAs) are small, noncoding RNAs that play essential roles in various stages during cancer progression. The aim of the current study was to elucidate the role of miR-1269 in the pathogenesis of HCC. METHODS: The expression of miR-1269 in HCC cells and tissues were determined by Real-time PCR analysis. Cell viability, colony formation and anchorage-independent growth ability assays were performed to examine cell proliferative capacity and tumorigenicity. Flow cytometry analysis was conducted to determine cell cycle progression. The expression of p21, CyclinD1, phosphorylated Rb, Rb and FOXO1 were examined by Western blotting analysis. Luciferase assay was used to determine whether FOXO1 is the direct target of miR-1269. RESULTS: miR-1269 was upregulated in HCC cells and tissues. Ectopic miR-1269 expression promoted, but inhibition of miR-1269 reduced, proliferation, tumorigenicity and cell cycle progression of HCC cells. Furthermore, we demonstrated that FOXO1 was a direct target of miR-1269. Suppression of FOXO1 by miR-1269 was associated with dysregulation of p21, cyclin D1, phosphorylated Rb and Ki67 expression, thereby playing an essential role in the growth of HCC cells. CONCLUSIONS: Our study indicated that overexpression of miR-1269 promotes cell proliferation in HCC through directly suppressing FOXO1, and functions as an oncomiR in HCC.

Improved accuracy of auto-segmentation of organs at risk in radiotherapy planning for nasopharyngeal carcinoma based on fully convolutional neural network deep learning.

OBJECTIVE: We examined a modified encoder-decoder architecture-based fully convolutional neural network, OrganNet, for simultaneous auto-segmentation of 24 organs at risk (OARs) in the head and neck, followed by validation tests and evaluation of clinical application. MATERIALS AND METHODS: Computed tomography (CT) images from 310 radiotherapy plans were used as the experimental data set, of which 260 and 50 were used as the training and test sets, respectively. An improved U-Net architecture was established by introducing a batch normalization layer, residual squeeze-and-excitation layer, and unique organ-specific loss function for deep learning training. The performance of the trained network model was evaluated by comparing the manual-delineation and the STAPLE contour of 10 physicians from different centers. RESULTS: Our model achieved good segmentation in all 24 OARs in nasopharyngeal cancer radiotherapy plan CT images, with an average Dice similarity coefficient of 83.75%. Specifically, the mean Dice coefficients in large-volume organs (brainstem, spinal cord, left/right parotid glands, left/right temporal lobes, and left/right mandibles) were 84.97% - 95.00%, and in small-volume organs (pituitary, lens, optic nerve, and optic chiasma) were 55.46% - 91.56%. respectively. Using the STAPLE contours as standard contour, the OrganNet achieved comparable or better DICE in organ segmentation then that of the manual-delineation as well. CONCLUSION: The established OrganNet enables simultaneous automatic segmentation of multiple targets on CT images of the head and neck radiotherapy plans, effectively improves the accuracy of U-Net based segmentation for OARs, especially for small-volume organs.

Integrating pretreatment MRI-detected nodal features and Epstein-Barr virus DNA to identify optimal candidates for intensity-modulated radiotherapy alone in patients with stage II nasopharyngeal carcinoma.

OBJECTIVES: To develop and validate a prognostic nomogram based on MRI-detected features of retropharyngeal and cervical lymph nodes and Epstein-Barr virus (EBV) DNA in patients with stage II nasopharyngeal carcinoma (NPC) to distinguish low-risk patients for whom intensity-modulated radiotherapy (IMRT) alone is sufficient. METHODS: This retrospective study enrolled 894 patients with stage II NPC (596 and 298 in the training and validation cohorts, respectively) with pretreatment MRI between August 2010 and May 2019. All patients received IMRT with or without additional chemotherapy. We identified independent risk factors using univariate and multivariate Cox regression analyses. Survival was compared using Kaplan-Meier curves with the log-rank test. RESULTS: Independent factors derived from the multivariate analysis include cervical nodal necrosis (CNN), the extracapsular spread (ECS) of cervical and retropharyngeal lymph nodes, and gamma-glutamyl transferase (γ-GGT). Nomograms A, B, and C were established based on the clinical [tumor-node-metastasis (TNM) stage + Epstein-Barr virus (EBV) DNA], the clinical-radiological [all independent predictors] and the combined models [the clinical-radiological model + EBV DNA], respectively. Nomogram C (C-index 0.769 [0.718-0.820]) demonstrated better risk discrimination than nomogram B (0.762 [0.715-0.809]), nomogram A (0.619 [0.564-0.674]), and the TNM stage (0.560 [0.509-0.611]). In the low-risk group divided by nomogram C, no significant survival differences were observed between patients treated with radiotherapy (RT) alone and other regimens including additional chemotherapy. CONCLUSIONS: The nomogram combining MRI-detected retropharyngeal and cervical lymph node features with pretreatment EBV-DNA improved the prognostic risk stratification for stage II NPC.

Multivariate NTCP Model of Hypothyroidism After Intensity-Modulated Radiotherapy for Nasopharyngeal Carcinoma.

OBJECTIVE: To evaluate the incidence of hypothyroidism in patients with nasopharyngeal carcinoma after intensity-modulated radiotherapy (IMRT), analyze its correlation with multiple influencing factors such as thyroid exposure dose, thyroid volume, and gender, and construct a multivariate-based normal tissue complication probability (NTCP) model for the occurrence of hypothyroidism after IMRT. MATERIALS AND METHODS: The thyroid hormone levels of patients at different points in time before and after radiotherapy were tested, and statistics on the incidence of hypothyroidism after treatment were obtained. The dose-volume data of patients' thyroids were converted into EQD2 equivalent dose values. The correlation between hypothyroidism after radiotherapy and thyroid exposure dose, thyroid volume, gender, and other factors was analyzed, and an NTCP model was constructed. RESULTS: A total of 69 patients with nasopharyngeal carcinoma were enrolled in this study. Twelve months after radiotherapy, a total of 24 patients (34.8%) developed hypothyroidism. Univariate analysis and multivariate analysis revealed that the average thyroid dose and thyroid volume are the most important factors affecting hypothyroidism after radiotherapy. The NTCP model constructed based on the average dose and thyroid volume has a good degree of fit. CONCLUSION: The volume and average dose of the thyroid gland are the key factors affecting the occurrence of hypothyroidism in patients with nasopharyngeal carcinoma after radiotherapy. The NTCP model constructed based on multivariate construction suggests that reducing the average dose of the thyroid to the greatest extent is an effective way to protect thyroid functions.

Case Report: Cetuximab in Combination With Chemotherapy for the Treatment of Multifocal Hepatic Metastases From Colorectal Cancer Guided by Genetic Tests.

Hepatic metastases were reported in up to 70% of colorectal cancer patients, among which multifocal hepatic metastasis represents one of the complications that lead to poor prognosis. The majority of the patients carrying multifocal hepatic metastases required pharmaceutical treatments to reduce the tumor size prior to surgical resection. However, the clinical responses to pharmaceutical agents were difficult to predict due to the heterogeneous nature of the multifocal tumors. Here, we report a case with multifocal hepatic metastases from colorectal cancer that was resistant to the primary chemotherapy and Bevacizumab plus chemotherapy, but responded to the combined therapy of Cetuximab and FOLFOX. Genetic tests had revealed that the tumor was highly metastatic due to the mutations of the WNT signaling pathway, and the metastatic tumors might be sensitive to Cetuximab. Consistent with the molecular characterizations, the metastatic tumors continue to emerge after chemotherapy, and rapidly relapsed in great numbers after liver resection. However, the combined therapy of Cetuximab and FOLFOX guided by the genetic tests significantly reduced the size and number of metastatic tumors. To conclude, deciphering the mutation profiles of multifocal metastatic tumors may guide the determination of treatment tactics, which may benefit the patients with non-resectable advanced carcinoma.

Computed Tomography-Based Evaluation of Volume and Position Changes of the Target Region and Organs at Risk During Radiotherapy for Esophageal Cancer: A Pilot Study.

OBJECTIVE: To analyze changes in volume and position of target regions and organs at risk (OARs) during radiotherapy for esophageal cancer patients. METHODS: Overall, 16 esophageal cancer patients who underwent radiotherapy, including 10 cases of intensity-modulated radiation therapy (IMRT) and six of three-dimensional conformal radiotherapy (3D-CRT), were enrolled. The prescription doses for the planning target volumes (PTVs) were as follows: PTV1, 64 Gy/32 fractions; and PTV2, 46 Gy/23 fractions. Repeat computed tomography (CT) was performed for patients after the 5th, 10th, 15th, 20th, and 25th fractions. Delineation of the gross tumor volume (GTV) and OAR volume was determined using five repeat CTs performed by the same physician. The target and OAR volumes and centroid positions were recorded and used to analyze volume change ratio (VCR), center displacement (ΔD), and changes in the distance from the OAR centroid positions to the planned radiotherapy isocenter (distance to isocenter, DTI) during treatment. RESULTS: No patient showed significant changes in target volume (TV) after the first week of radiotherapy (five fractions). However, TV gradually decreased over the following weeks, with the rate slowing after the fourth week (40 Gy). The comparison of TV from baseline to 40 Gy (20 fractions) showed that average GTVs decreased from 130.7 ± 63.1 cc to 92.1 ± 47.2 cc, with a VCR of -29.21 ± 13.96% (p<0.01), while the clinical target volume (CTV1) decreased from 276.7 ± 98.2 cc to 246.7 ± 87.2 cc, with a VCR of -10.34 ± 7.58% (p<0.01). As TVs decreased, ΔD increased and DTI decreased. After the fourth week of radiotherapy (40 Gy), centroids of GTV, CTV1, and prophylactic CTV (CTV2) showed average deviations in ΔD of 7.6 ± 4.0, 6.9 ± 3.4, and 6.0 ± 3.0 mm, respectively. The average DTI of the heart decreased by 4.53 mm (from 15.61 ± 2.96 cm to 15.16 ± 2.27 cm). CONCLUSION: During radiotherapy for esophageal cancer, Targets and OARs change significantly in volume and position during the 2nd-4th weeks. Image-guidance and evaluation of dosimetric changes are recommended for these fractions of treatment to appropriate adjust treatment plans.

Interobserver variations in the delineation of target volumes and organs at risk and their impact on dose distribution in intensity-modulated radiation therapy for nasopharyngeal carcinoma.

OBJECTIVE: This study aimed to (a) assess the differences in the delineation of target volumes and organs-at-risk (OARs) by different physicians designing an intensity-modulated radiation therapy (IMRT) for nasopharyngeal carcinoma (NPC) and (b) analyze the impact of these differences on the treatment plan optimization. MATERIALS AND METHODS: The planning target volumes (PTVs) and OARs for radiotherapy were manually delineated from computed tomography images of a patient with NPC, and a standard delineation was determined using the STAPLE algorithm of ABAS software. IMRT was designed using one standard plan and 10 individual plans based on the same constraints and field conditions. The maximum/minimum ratio (MMR) of the PTV and OAR volumes and the coefficient of variation (CV) for the different groups were evaluated and compared to the volume of the standard contour. RESULTS: Significant differences were seen in the PTVs of the nasopharynx (PTVnx), neck lymph node (PTVnd) and the OARs manually delineated by different physicians. Compared to the standard plan, the mean dose-related parameters of various OARs in different individual plans were not significantly different, while that of most organs in different individual plans were reduced. However, a significant difference in the dose at each organ was noted in different individual plans. CONCLUSION: Significant differences were noted in the PTV and OAR delineations by different physicians in radiotherapy of NPC, and their dosimetric parameters were significantly different from the standard planned parameters. Therefore, multicenter trials should pay attention to the impact of these differences on the clinical evaluation.

Advantage of PET/CT in Target Delineation of MRI-negative Cervical Lymph Nodes In Intensity-Modulated Radiation Therapy Planning for Nasopharyngeal Carcinoma.

Introduction: In intensity-modulated radiation therapy (IMRT) planning for nasopharyngeal carcinoma (NPC), cervical lymph nodes (CLNs) that appear negative on magnetic resonance imaging (MRI) scans can be difficult to target. The purpose of this study was to assess the advantage of 18F-fluorodeoxyglucose positron emission tomography with computed tomography (18F-FDG PET/CT) for distinguishing MRI-negative CLNs and the effect of 18F-FDG PET/CT on diagnosis, target delineation, and dose prescription in IMRT planning for NPC. Methods: Thirty-five NPC patients with 37 MRI-negative CLNs underwent 18F-FDG PET/CT imaging before treatment. Ultrasonography-guided fine-needle aspiration cytology (USgFNAC) was performed to examine the pathology of CLNs. The 18F-FDG PET/CT and cytopathological results were compared, and the diagnostic accuracy of 18F-FDG PET/CT was calculated. The cytopathologically confirmed CLNs were delineated and treated as the gross tumor volume of lymph nodes (denoted as GTVnd). Results: Nineteen of the 37 MRI-negative CLNs were positive on 18F-FDG PET/CT, and metastasis was confirmed by USgFNAC in 16 CLNs. Of the remaining 18 18F-FDG PET/CT-negative lymph nodes, metastasis was confirmed in one. The diagnostic accuracy, sensitivity, and specificity of 18F-FDG PET/CT were 89.2%, 94.1%, and 85.0%, respectively. The positive and negative predictive values were 84.2% and 94.4%, respectively. With a median follow-up of 48.3 months, no relapse was observed among the 18F-FDG PET/CT-positive CLNs with metastasis confirmed by USgFNAC and treated as GTVnd. Conclusion:18F-FDG PET/CT had high accuracy, sensitivity, and specificity for distinguishing MRI-negative CLNs. 18F-FDG PET/CT-positive CLNs could reasonably be categorized as high-risk clinical tumor volume in IMRT planning for NPC.

Development and validation of a nomogram for predicting the survival of patients with non-metastatic nasopharyngeal carcinoma after curative treatment.

BACKGROUND: The TNM staging system is far from perfect in predicting the survival of individual cancer patients because only the gross anatomy is considered. The survival rates of the patients who have the same TNM stage disease vary across a wide spectrum. This study aimed to develop a nomogram that incorporates other clinicopathologic factors for predicting the overall survival (OS) of non-metastatic nasopharyngeal carcinoma (NPC) patients after curative treatments. METHODS: We retrospectively collected the clinical data of 1520 NPC patients who were diagnosed histologically between November 2000 and September 2003. The clinical data of a separate cohort of 464 patients who received intensity-modulated radiation therapy (IMRT) between 2001 and 2010 were also retrieved to examine the extensibility of the model. Cox regression analysis was used to identify the prognostic factors for building the nomogram. The predictive accuracy and discriminative ability were measured using the concordance index (c-index). RESULTS: We identified and incorporated 12 independent clinical factors into the nomogram. The calibration curves showed that the prediction of OS was in good agreement with the actual observation in the internal validation set and IMRT cohort. The c-index of the nomogram was statistically higher than that of the 7th edition TNM staging system for predicting the survival in both the primary cohort (0.69 vs. 0.62) and the IMRT cohort (0.67 vs. 0.63). CONCLUSION: We developed and validated a novel nomogram that outperformed the TNM staging system in predicting the OS of non-metastatic NPC patients who underwent curative therapy.

Positron emission tomography-computed tomography before treatment is highly prognostic of distant metastasis in nasopharyngeal carcinoma patients after intensity-modulated radiotherapy treatment: a prospective study with long-term follow-up.

OBJECTIVES: The utility of [18F]fluorodeoxyglucose (FDG) positron emission tomography (PET) in the prediction of distant metastasis after definitive treatment for non-disseminated NPC patients is not clear. We prospectively investigated the prognostic significance of (18)F-FDG PET-CT performed before treatment for the distant metastasis-free survival (DMFS) of NPC patients treated with intensity-modulated radiotherapy (IMRT) ± chemotherapy. MATERIALS AND METHODS: The results of PET-CT scans performed at initial diagnosis were recorded and analyzed prospectively. Patients then received standard treatment per protocol independent of the PET-CT result. IMRT was administered to the nasopharynx and neck. Early stage patients received IMRT alone, whereas loco-regionally advanced patients received concurrent chemoradiation. Patient outcomes were evaluated. RESULTS: One hundred and seventy-nine patients were analyzable. DM occurred in 33 patients and the 3-year DMFS rate of the whole cohort was 84.1%. The cut-off of the SUVmax at the primary site (SUVmax-P) for DMFS was 10.22 by ROC curve. The 3-year DMFS rates of the higher and lower SUVmax-P groups (SUVmax-P⩽or>10.22) were 93.2% and 75.1%, respectively, with an 18.1% difference (P=0.003). Multivariate analysis indicated that SUVmax-P was a risk factor independently associated with DMFS (HR, 2.672; 95%CI, 1.236-5.776; P=0.012) and OS (HR, 2.417; 95%CI, 1.313-4.448; P=0.005). Subgroup analysis indicated that the 5-year DMFS and OS in loco-regionally advanced patients with SUVmax-P⩽10.22 were similar to those of early stage patients, whereas those of loco-regionally advanced patients with SUVmax-P>10.22 patients predicted worse outcome. CONCLUSION: SUVmax-P is a useful biomarker to predict distant metastasis of NPC patients treated with IMRT. Combining SUVmax-P with tumor overall stage, a more precise picture could be obtained to predict treatment outcome.

Cathepsin L is involved in proliferation and invasion of ovarian cancer cells.

Cathepsin L (CTSL) is a lysosomal cysteine protease that has been found to be overexpressed in ovarian cancer (OC). The aim of the present study was to investigate the possible involvement of CTSL in the development of OC. In this study, RNA interference with a CTSL small hairpin RNA (CTSL-shRNA), and a plasmid carrying CTSL were used to identify the effects of this enzyme on the regulation of the malignant behavior of OC cells. OV-90 and SKOV3 human ovarian cancer cell lines were selected as cell models in vitro and in vivo. The results showed that downregulation of CTSL significantly inhibits the proliferative and invasive capability of SKOV3 cells, and that upregulation of CTSL in OV-90 cells leads to opposite effects. Compared with parental OC cells, cells in which CTSL was silenced exhibited a reduced capacity to develop into tumors in nude mice, while the growth of tumor xenografts derived from these cells was markedly constrained. In conclusion, the results suggested that CTSL contributes to the proliferation and metastasis of OC, and that CTSL may be a novel molecular target for OC treatment.

Distant metastasis risk and patterns of nasopharyngeal carcinoma in the era of IMRT: long-term results and benefits of chemotherapy.

PURPOSE: To report the distant metastasis (DM) risk and patterns for nasopharyngeal carcinoma (NPC) treated with intensity-modulated radiotherapy (IMRT) and to analyze the benefits of chemotherapy based on DM risk. MATERIALS AND METHODS: 576 NPC patients were analyzed. The DM rates were calculated using the Kaplan-Meier method, and the log-rank test was used to compare differences. The patients were divided into different risk subclassifications according to DM hazard ratios. RESULTS: 91 patients developed DM after treatment, with bone as the most common metastatic sites. 82.4% of DMs occurred within 3 years of treatment. Patients were classified as low-risk, intermediate-risk and high-risk, and the corresponding 5-year DM rates were 5.1%, 13.1% and 32.4%, respectively (P < 0.001). Chemotherapy failed to decrease the DM rate in the low-risk subclassification, but decreased the DM risk in the intermediate-risk subclassification (P = 0.025). In the high-risk subclassification, the DM rate was 31.9% though chemotherapy was used, which was significantly higher than that of other two subclassifications. CONCLUSIONS: DM is the dominant treatment failure in NPC treated by IMRT, with similar occurrence times and distributions to those that occurred in the era of conventional radiotherapy. Further studies on treatment optimization are needed in high-risk patients.