TMEM189 promotes breast cancer through inhibition of autophagy-regulated ferroptosis.

Breast cancer is a leading cause of tumor-related death among women around the world, but its pathogenesis is still unclear. Transmembrane protein 189 (TMEM189) is widely expressed in many types of tissues and plays a critical role in tumorigenesis partly through mediating cell death. However, its regulatory function on breast cancer progression and particularly the underlying mechanisms have not been fully understood. In the present study, we found that TMEM189 knockdown significantly reduced the proliferation of breast cancer cells, while its over-expression facilitated the proliferative capacity of tumor cells. The effects of TMEM189 to promote breast cancer were validated in the constructed xenograft mouse models. RNA-sequencing studies subsequently showed that TMEM189 deletion was closely associated with ferroptosis signaling pathway, accompanied with elevated lipid reactive oxygen species (ROS) accumulation, cellular ROS production, malondialdehyde (MDA) and the intracellular iron releases. However, GSH levels in breast cancer cells were highly impeded upon TMEM189 inhibition. Intriguingly, we found that TMEM189 knockdown-induced ferroptotic cell death was considerably abolished after autophagy inhibitor 3-MA co-treatment, as evidenced by the markedly decreased ROS generation and intracellular iron accumulation. Moreover, TMEM189 ablation strongly up-regulated LC3BII and transferrin receptor 1 (TfR1) expression levels in breast cancer cells, whereas down-regulated p62 and GPX4. Importantly, the expression changes of these molecules related to autophagy and ferroptosis were almost diminished in response to 3-MA exposure, along with restored cell proliferation. These findings suggested that TMEM189 could inhibit autophagy to mediate ferroptosis in breast cancer cells. Collectively, all our findings revealed the therapeutic potential of TMEM189 in the treatment of breast cancer.

Mutational landscape of nasopharyngeal carcinoma based on targeted next-generation sequencing: implications for predicting clinical outcomes.

BACKGROUND: The aim of this study was to draw a comprehensive mutational landscape of nasopharyngeal carcinoma (NPC) tumors and identify the prognostic factors for distant metastasis-free survival (DMFS). METHODS: A total of forty primary nonkeratinizing NPC patients underwent targeted next-generation sequencing of 450 cancer-relevant genes. Analysis of these sequencing and clinical data was performed comprehensively. Univariate Cox regression analysis and multivariate Lasso-Cox regression analyses were performed to identify factors that predict distant metastasis and construct a risk score model, and seventy percent of patients were randomly selected from among the samples as a validation cohort. A receiver operating characteristic (ROC) curve and Harrell's concordance index (C-index) were used to investigate whether the risk score was superior to the TNM stage in predicting the survival of patients. The survival of patients was determined by Kaplan-Meier curves and log-rank tests. RESULTS: The twenty most frequently mutated genes were identified, such as KMT2D, CYLD, and TP53 et al. Their mutation frequencies of them were compared with those of the COSMIC database and cBioPortal database. N stage, tumor mutational burden (TMB), PIK3CA, and SF3B1 were identified as predictors to build the risk score model. The risk score model showed a higher AUC and C-index than the TNM stage model, regardless of the training cohort or validation cohort. Moreover, this study found that patients with tumors harboring PI3K/AKT or RAS pathway mutations have worse DMFS than their wild-type counterparts. CONCLUSIONS: In this study, we drew a mutational landscape of NPC tumors and established a novel four predictor-based prognostic model, which had much better predictive capacity than TNM stage.

18F-fluorodeoxyglucose positron emission computed tomography for monitoring tumor response in esophageal carcinoma treated with concurrent chemoradiotherapy.

The aim of the present study was to explore the value of 18F-fluorodeoxyglucose positron emission tomography (18F-FDG PET) in monitoring the early tumor response of esophageal squamous cell carcinoma (ESCC) treated with concurrent chemoradiotherapy (CRT). A total of 48 patients with pathologically proven ESCC were retrospectively analyzed. All patients underwent two serial 18F-FDG PET scans at baseline (pre-CRT) and 40 Gy/4 weeks of starting radiation therapy (inter-CRT). All patients received intensity-modulated radiotherapy (with a total radiation dose of 59.6 Gy) concurrently with cisplatin-based chemotherapy. The maximum standardized uptake value (SUVmax) and metabolic tumor volume (MTV) were measured using 18F-FDG PET. The percentage changes (Δ) in SUVmax and MTV between two serial scans were calculated and were revealed to be associated with the objective tumor response (oTR), according to the Response Evaluation Criteria in Solid Tumors 1.1. Among the 48 patients, 20.8% achieved a complete response, 68.8% exhibited a partial response and the oTR rate was 89.6%. On the pre-CRT PET scans, the mean SUVmax and MTV were 14.1±5.8 and 58.2±25.4 cm3, respectively. Following 40 Gy irradiation over 4 weeks, the mean SUVmax and MTV significantly decreased to 4.3±3.5 and 19.0±12.1 cm3, respectively (P<0.001). A significantly higher ΔSUVmax and ΔMTV was observed in the responders compared with that in the non-responders [0.71±0.16 vs. 0.51±0.26 (P=0.015); and 0.64±0.13 vs. 0.42±0.09 (P=0.001), respectively]. Univariate analysis revealed that ΔSUVmax and ΔMTV were significantly associated with oTR (P=0.010 and P=0.001, respectively). ΔMTV was used as a predictor and a cut-off value of 54% discriminated responders from non-responders with a sensitivity of 69.8% and a specificity of 100% (P=0.001). The area under the receiver operating characteristic curve was 0.837 (95% confidence interval, 0.702-0.928). The results of the present study indicated that interim 18F-FDG PET scans may provide early prognostic value for determining oTR in patients with ESCC undergoing treatment with CRT.

Which is the optimal threshold for defining functional lung in single-photon emission computed tomography lung perfusion imaging of lung cancer patients?

PURPOSE: The aim of this study was to investigate the optimal threshold for the functional lung (FL) definition of single-photon emission computed tomography (SPECT) lung perfusion imaging. PATIENTS AND METHODS: Forty consecutive stage III non-small-cell lung cancer patients underwent SPECT lung perfusion scans and PET/CT scans for treatment planning, and the images were coregistered. Total lung and perfusion lung volumes corresponding to 10, 20, …, 60% of the maximum SPECT count were segmented automatically. The SPECT-weighted mean lung dose (SWMDx%) and the percentage of FL volume receiving more than 20 Gy (Fx%V20) of different thresholds were investigated using SPECT-weighted dose-volume histograms. Receiver-operator characteristic curves were used to identify SWMD and FV20 of different thresholds in predicting the incidence of radiation pneumonitis (RP). RESULTS: Eleven (27.5%) patients developed RP (grades 1, 2, 3, and 4 were 10.0, 7.5, 7.5, and 2.5%, respectively) after treatment. The largest area under the receiver-operator characteristic curve was 0.881 for the ability of SWMD to predict RP with 20% as the threshold and 0.928 for the ability of FV20 with 20% as the threshold. CONCLUSION: The SWMD20% and FV20 of FL using 20% of the maximum SPECT count as the threshold may be better predictors for the risk of RP.

Clinical significance of serum dynamics of HSP90a level in esophageal squamous cell carcinoma patients treated with definitive chemoradiotherapy.

PURPOSE: We evaluate whether the change of heat shock protein 90a (HSP90a) level before and after definitive chemoradiotherapy (CRT) in esophageal squamous cell carcinoma (ESCC) affects tumor response and overall survival (OS). This study aimed to investigate the role of HSP90a reduction ratio after CRT. METHODS: Correlations between pre-CRT HSP90a levels and the tumor response to CRT were analysed. Patients were divided into three groups (Group 1: Serum HSP90a levels pretreatment CRT less than 124 ng/mL; Group 2: pre-CRT HSP90a of 124 ng/mL or more with HSP90a reduction ratio of 65% or more; Group 3: pre-CRT HSP90a of 124 ng/mL or more with HSP90a reduction ratio less than 65%), and their oncologic outcomes were compared. RESULTS: The rates of good response in HSP90a low (pre-CRT HSP90a ≤ 124 ng/mL) and high groups (pre-CRT HSP90a ≤ 124 ng/mL) were 67.3% (68/101) and 37.78% (20/79), respectively (P= 0.004). The rates of good response were significantly higher in Group 1 than in Groups 2 and 3 (58.5% vs. 46.0% and 27.8%, respectively; P= 0.013). The results from statistical analysis indicated that the tumor response was significantly associated with the serum levels of pre-CRT HSP90a and HSP90a Group (P< 0.05). The OS rate was not different between Groups 1 and 2 but was significantly lower in Group 3. HSP90a Group were independent prognostic factors for OS. CONCLUSIONS: HSP90a levels could be of clinical value as a predictor of response to CRT HSP90a reduction ratio might be an independent prognostic factor for in ESCC patients treated with definitive CRT.

Management of salivary gland carcinomas - a review.

Salivary gland carcinomas are a heterogeneous group of tumors with many histological subtypes which occur in both major and minor salivary glands. However, they have a relatively low of incidence. Their rarity limits study size and the ability to perform phase III trials. Therefore, to date, the entire management is usually varied. Certain published studies have paid more attention to the systemic therapy in the management of metastatic or locally recurrent salivary gland cancer, while little effort has been made to study the entire management for this lesions. Although results of treatment for patients with salivary gland carcinoma have improved in recent years, the treatment of salivary gland cancers is still not standardized. And some patients who haven't received optimal treatment strategies had a reduced survival. In this review, the topics covered include surgery and radiotherapy, selective neck dissection, chemotherapy, and targeted therapy, which aimed to summarize the optimal management approaches and to develop recommendations for managing this lesions. For these rare cancers, there is also a need for a determined, coordinated effort to conduct high-quality clinical trials.

Anatomic distribution of supraclavicular lymph node in patients with esophageal cancer.

PURPOSE: Definitive chemoradiation therapy remains the standard of care for patients with localized esophageal carcinoma who choose nonsurgical management. However, there is no consensus regarding delineation of the nodal clinical target volume (CTVn), especially for lower cervical lymph nodes. This study aimed to map the location of metastatic supraclavicular lymph nodes in thoracic esophageal carcinoma patients with supraclavicular node involvement and generate an atlas to delineate the CTVn for elective nodal radiation of esophageal squamous cell carcinoma. PATIENTS AND METHODS: In this study, the supraclavicular regional lymph node was further divided into four subgroups. The locations of the involved supraclavicular nodes for all patients were then transferred onto a template computed tomography (CT) image. A volume probability map was then generated with nodal volumes, and was displayed on the template CT to provide a visual impression of nodal frequencies and anatomic distribution. RESULTS: We identified 154 supraclavicular nodal metastases based on CT image in 96 patients. Of these, 29.2% were located in group I region, 59.7% in group II region, 10.4% in group III region, and 0.7% in group IV region. CONCLUSION: On the basis of our study, we suggest that the appropriate radiation field of CTVn should include the group I and II regions and the CTVn exterior margin along the lateral side of the internal jugular vein may be suitable.

Recurrence pattern of squamous cell carcinoma in the midthoracic esophagus: implications for the clinical target volume design of postoperative radiotherapy.

BACKGROUND: Postoperative radiotherapy has shown positive efficacy in lowering the recurrence rate and improving the survival rate for patients with esophageal squamous cell carcinoma (ESCC). However, controversies still exist about the postoperative prophylactic radiation target volume. This study was designed to analyze the patterns of recurrence and to provide a reference for determination of the postoperative radiotherapy target volume for patients with midthoracic ESCC. PATIENTS AND METHODS: A total of 338 patients with recurrent or metastatic midthoracic ESCC after radical surgery were retrospectively examined. The patterns of recurrence including locoregional and distant metastasis were analyzed for these patients. RESULTS: The rates of lymph node (LN) metastasis were 28.4% supraclavicular, 77.2% upper mediastinal, 32.0% middle mediastinal, 50.0% lower mediastinal, and 19.5% abdominal LNs. In subgroup analyses, the rate of abdominal LN metastasis was significantly higher in patients with histological node-positive than that in patients with histological node-negative (P=0.033). Further analysis in patients with histological node-positive demonstrated that patients with three or more positive nodes are more prone to abdominal LN metastasis, compared with patients with one or two positive nodes (χ2=4.367, P=0.037). The length of tumor and histological differentiation were also the high-risk factors for abdominal LN metastasis. CONCLUSION: For midthoracic ESCC with histological node-negative, or one or two positive nodes, the supraclavicular and stations 2, 4, 5, and 7 LNs should be delineated as clinical target volume of postoperative prophylactic irradiation, and upper abdominal LNs should be excluded. While for midthoracic ESCC with three or more positive nodes, upper abdominal LNs should also be included. The length of tumor and histological differentiation should be considered comprehensively to design the clinical target volume for radiotherapy.

Genetic variants in inducible nitric oxide synthase gene are associated with the risk of radiation-induced lung injury in lung cancer patients receiving definitive thoracic radiation.

BACKGROUND AND PURPOSE: Nitric oxide (NO), mainly synthesized by inducible nitric oxide synthase (NOS2) in pathological conditions, plays an important role in cytotoxicity, inflammation and fibrosis. Elevations in exhaled NO after thoracic radiation have been reported to predict radiation-induced lung injury (RILI). This study examined whether genetic variations in NOS2 gene is associated with the risk of RILI. MATERIAL AND METHODS: A cohort of 301 patients between 2009 and 2011 were genotyped for 21 single nucleotide polymorphisms (SNPs) in the NOS2 gene by the Sequenom MassArray system. Kaplan-Meier cumulative probability was used to assess RILI risk and Cox proportional hazards analyses were performed to evaluate the effect of NOS2 genotypes on RILI. RESULTS: Multivariate analysis found that three SNPs (rs2297518, rs1137933 and rs16949) in NOS2 were significantly associated with risk of RILI⩾2 (P value=0.001, 0.000092, 0.001, respectively) after adjusting for other covariates. Their associations were independent of radiation dose and mean lung dose. Further haplotype analysis indicated that the ATC haplotype of three SNPs is associated with reducing the risk of developing RILI. CONCLUSION: Our results demonstrate that genetic variants of NOS2 may serve as a reliable predictor of RILI in lung cancer patients treated with thoracic radiation.

Long-term outcome of sensorineural hearing loss in nasopharyngeal carcinoma patients: comparison between treatment with radiotherapy alone and chemoradiotherapy.

The purpose of this study is to assess the long-term effect of sensorineural hearing loss (SNHL) resulted from radiotherapy (RT) alone versus chemoradiotherapy in nasopharyngeal carcinoma patients (NPC). Seventy-two patients initially diagnosed with NPC were enrolled from Shandong Tumor Hospital between March 2003 and May 2007. They were assigned into two groups: RT alone and chemoradiotherapy according to the different treatment regimens. Intensity-modulated radiation therapy was applied for both groups, concurrent and adjuvant cisplatin were administered for chemoradiotherapy group additionally. Hearing threshold test was performed at various time periods after completion of RT. Mean radiation dose to the cochlea in each ear was calculated to determine the correlation between cochlear dose and SNHL. We found that the hearing loss is more severe in the chemoradiotherapy group compared with RT group, from completion of RT up to the 5 years of follow-up period. This is especially obvious in the high frequency range. Hearing level is seriously damaged when cochlea dose exceeds 46 GY. We concluded that concurrent/adjuvant chemotherapy plus RT aggravates SNHL in NPC patients than RT alone and thus inner ear tissue tolerance should be redefined in those patients.

Elective lymph node irradiation late course accelerated hyper-fractionated radiotherapy plus concurrent cisplatin-based chemotherapy for esophageal squamous cell carcinoma: a phase II study.

BACKGROUND: In this phase II study, we evaluated the efficacy, toxicity, and patterns of failure of elective lymph node irradiation (ENI) late course accelerated hyper-fractionated radiotherapy (LCAHRT) concurrently with cisplatin-based chemotherapy (CHT) for esophageal squamous cell carcinoma (ESCC). METHODS: Patients with clinical stage II-IVa (T1-4N0-1M0 or M1a) ESCC were enrolled between 2004 and 2011. Radiation therapy (RT) comprised two courses: The first course of radiation covered the primary and metastatic regional tumors and high risk lymph nodal regions, given at 2 Gy per fraction for a dose of 40 Gy. In the second course, LCAHRT was delivered to the boost volume twice a day for an additional 19.6 Gy in 7 treatment days, using 1.4 Gy per fraction. Two cycles of CHT were given at the beginning of RT. RESULTS: The median age and Karnofsky performance status were 63 years and 80, respectively. The American Joint Committee on Cancer stage was II in 14 (20.6%) patients, III in 32 (47.1%), and IVa in 22 (32.3%). With a median follow-up of 18.5 months, the overall survival at 1-, 3-, 5-year were 75.5%, 46.5%, 22.7% for whole group patients, versus 78.6%, 49.4%, 39.9% for patients with stage II-III. The patterns of first failure from local recurrence, regional failure, and distant metastasis were seen in 20.6%, 17.6%, and 19.1%, respectively. The most frequent acute high-grade (≥ 3) toxicities were esophagitis and leucopenia, occurred in 26.4% and 32.4%. CONCLUSIONS: ENI LCAHRT concurrently with CHT was appeared to be an effective regimen for ESCC patient with a favorable and tolerated profile. Further observation with longer time and randomized phase III trial is currently underway. TRIAL REGISTRATION: ChiCTR-TRC-09000568.

Spect-guidance to reduce radioactive dose to functioning lung for stage III non-small cell lung cancer.

OBJECTIVE: To investigate the treatment effect of additional information obtained by single photon emission computed tomography (SPECT) lung perfusion imaging (LPI) in the radiotherapy planning process for patients with stage III non-small cell lung cancer (NSCLC). METHODS: 39 patients with stage III NSCLC were enrolled. Gross tumor volume (GTV) was outlined by SPECT/CT images, SPECT-LPIs being used to define functional lung (FL) and non-functional lung (NFL) regions. Two sets of IMRT plans were designed to deliver 64Gy to PTV. One was a regular IMRT plan using CT images only (Plan 1), and the other was a corresponding IMRT plan using co-registered images (Plan 2). FLVx (the % volume of functional lung receiving ≥x Gy) and WLVx (% volume of whole lung to receive ≥x Gy) were compared by paired Student's t test. Kendalls correlation was used to analyze the factor (s) related with the FLV20 decrease. RESULTS: Compared with plan 1, both WLVx and FLVx were decreased in plan 2. WLV10, WLV15, WLV20, WLV25, WLV30 and WLV35 decreased 9.7%, 13.8%, 17.2%, 12.9%, 9.8% and 9.8%, and FLV10, FLV15, FLV20, FLV25, FLV30 and FLV35 decreased 10.8%, 14.6%, 17.3%, 14.5%, 14.5% and 10.5%. FLVx decreased significantly compared with WLVx. There were significant differences in WLV10, WLV15, WLV20, WLV25, WLV3 and FLV10, FLV15, FLV20, FLV25, FLV30 between plan 1 and plan 2 (P=0.002, 0.000, 0.000, 0.005, 0.027 and 0.002, 0.000, 0.000, 0.006, 0.010). According to Kendall correlation analysis, NFL had a negative relation with the percentage FLV20 decrease (r=-0.559, P<0.01), while the distance of PTV and NFL center had a significantly positive relation with the percentage of FLV20 decrease (r=0.768, P<0.01). CONCLUSION: Routine use of SPECT-LPI for patients undergoing radiotherapy planning for stage III NSCLC appears warranted.

Significant associations between GSTM1/GSTT1 polymorphisms and nasopharyngeal cancer risk.

Glutathione S-transferases play a critical role in the detoxification and elimination of electrophilic carcinogens by conjugating them to glutathione. Homozygous deletions of GSTM1 and GSTT1 have been suggested as risk factors for some cancers, including colorectal, pancreatic, and esophageal cancers. Results of previous individual studies published to estimate the associations between GSTM1/GSTT1 polymorphisms and nasopharyngeal cancer (NPC) risk remained controversial. Thus, we carried out a meta-analysis by pooling the odds ratios (ORs) with corresponding 95 % confidence intervals (95 % CIs) of all currently available case-control studies to shed some light on the contradictory finding. A comprehensive search of the PubMed, Embase, Web of Science, and China National Knowledge Infrastructure databases up to October 20, 2012 was performed to identify eligible studies. A total of 15 separate publications involving 2,226 NPC cases and 3,339 controls were finally included into this meta-analysis. The meta-analysis of total studies showed that the null genotypes of GSTM1 and GSTT1 were both significantly associated with increased risk of NPC (for GSTM1: OR = 1.54, 95 % CI 1.28-1.86, P OR < 0.001; for GSTT1: OR = 2.25, 95 % CI 1.50-3.36, P OR < 0.001). Subgroup analysis by ethnicity suggested that carriers of both GSTM1 and GSTT1 null genotypes in Asians were more susceptible to NPC. Additionally, in the subgroup analysis based on the sample size, significant associations of the GSTM1 and GSTT1 polymorphisms with NPC susceptibility were identified among studies both with larger case sample size (number of cases ≥ 100) and smaller case sample size (number of cases <100). Sensitivity analysis confirmed the stability of our results. These results indicate that the GSTM1 and GSTT1 polymorphisms may play crucial roles in the development of NPC, especially in Asians.

Relationship between (18)F-fluorodeoxyglucose uptake in primary lesions and clinicopathological characteristics of esophageal squamous cell carcinoma patients.

The aim of this study, was to investigate the relationship between (18)F-fluorodeoxyglucose ((18)F-FDG) uptake in primary tumors and the clinicopathological characteristics of esophageal squamous cell carcinoma (ESCC) patients. Patients with histopathologically diagnosed ESCC who had received a pre-therapeutic (18)F-FDG positron emission tomography-computed tomography (PET-CT) scan were enrolled in the study. The maximum standardized uptake value (SUVmax) and the length of the primary tumor were measured by PET-CT. The clinical tumor-node-metastasis (TNM) stage was determined mainly by PET-CT images according to the American Joint Committee on Cancer (AJCC) staging system, 2002. A significant difference was observed in SUVmax between the length and T stage of the primary tumor (P=0.000 and P=0.017, respectively), but not in the grade of tumor differentiation (P=0.383), clinical stage (P=0.583), N staging (P=0.387), M staging (P=0.886), patient age (P= 0.752) or gender (P=0.233). There was a significant positive correlation between the SUVmax and the length of the tumor (r=0.456, P=0.000) and the depth of invasion of the primary tumor (r=0.257, P=0.006). After controlling for length, no statistically significant correlation was found between T stage and SUVmax (r=0.074, P=0.537). In conclusion, these findings suggest that tumor length influences FDG uptake in ESCC tumors, and that the T stage of the primary tumor is not significantly correlated with the SUVmax after controlling for length. However, we did not find a significant correlation between the SUVmax and primary tumor differentiation and clinical stage. These data provide important information for the management of ESCC.

Serum superoxide dismutase, a potential predictor for radiation pneumonitis following chemoradiotherapy in non-small cell lung cancer patients.

PURPOSE: To explore serum superoxide dismutase (SOD) for predicting radiation pneumonitis (RP) in non-small cell lung cancer patients following chemoradiotherapy. METHODS: Serum levels for SOD were measured by enzyme-linked immunosorbent assays prior to radiation therapy (Pre-RT) and post 40 Gy/4 weeks during the treatment (Pos-RT). RESULTS: SOD concentrations after delivery of 40 Gy/4 weeks was associated with the development of RP. The best predictive ability of SOD was observed for a cut-off value of 56 unit/ml, with a sensitivity of 0.80 (95% CI 0.28-0.99), and a specificity of 0.67 (95% CI 0.43-0.65) (p = 0.040). CONCLUSION: Serum SOD may be a potential predictor for RP, which need to be further verified.