Effect of benzo[a]pyrene on proliferation and metastasis of oral squamous cell carcinoma cells: A transcriptome analysis based on RNA-seq.

Benzo[a]pyrene (BaP), a representative polycyclic aromatic hydrocarbon compound, is a carcinogen that causes head and neck cancers. Despite intensive research, the molecular mechanism of BaP in the development of oral squamous cell carcinoma (OSCC) remains largely unknown. In the present study, the SCC-9 human OSCC cell line was cultured in vitro, separated into treatment groups, and treated with dimethyl sulfoxide or BaP at various concentrations. The malignant behavior ascribed to the BaP treatment was investigated by cell proliferation, clony formation assay, and Transwell assays. Furthermore, transcriptome sequencing was performed to detect the differentially expressed genes, followed by quantitative real-time PCR to measure the expression levels of nine of these genes. Moreover, the Gene Ontology (GO) term and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed the biological processes and signaling pathways in which the target genes were involved. Significant effects on SCC-9 cell proliferation, tumorigenicity, cell migration, and invasion were observed after exposure to 8 µM BaP. Additional results revealed that BaP inhibited apoptosis in a dose-dependent manner. The transcriptome sequencing results showed 137 upregulated genes and 135 downregulated genes induced by BaP, associated with tumor-related biological processes and signaling pathways, mainly including transcriptional dysregulation in cancer, the tumor necrosis factor signaling pathway, metabolism of xenobiotics by cytochrome P450, mitogen-activated protein kinase signaling pathway, and so forth. Our study demonstrates that BaP may regulate the expression of certain genes involved in tumor-associated signaling pathways, thereby promoting the proliferative, tumorigenic, and metastatic behaviors of OSCC cells while suppressing their apoptosis.

Synergistic effects of autophagy inhibitors combined with cisplatin against cisplatin-resistant nasopharyngeal cancer cells.

This study explored the synergistic effects of autophagy inhibitors combined with cisplatin against cisplatin-resistant nasopharyngeal cancer cells by treating HNE-1 and cisplatin (diamminedichloroplatinum; DDP)-resistant HNE1/DDP nasopharyngeal cancer cell lines with DDP, autophagy inhibitors, or a combination of autophagy inhibitors and DDP. Cell viability was determined via MTT (colorimetric) and colony-forming assays, and the rate of apoptosis was determined using propidium iodide (PI) and annexin V double-staining. The expressions of proteins were determined by Western blotting. For our in-vivo studies, a murine xenograft model was established to evaluate the anti-tumor effects of the combination of autophagy inhibitor and DDP. The results showed that treatment with DDP increased the expressions of ATP-binding cassette sub-family B member 1 (ABCB1), ATP Binding Cassette Subfamily C Member 1 (ABCC1), and P-glycoprotein 1 (P-gp) in the HNE1/DDP cell lines. Treatment with chloroquine decreased the expression levels of ABCB1, ABCC1, and P-gp, and increased the formation of LC3-II and the expression levels of p62 in the HNE1/DDP cells. Additionally, the combination of autophagy inhibitors and DDP produced a synergistic effect on DDP-induced cell death and apoptosis. Furthermore, the combination of the autophagy inhibitor and DDP showed significant anti-tumor effects in the xenograft mouse model. In summary, autophagy inhibitors show synergistic anti-tumor effects with DDP in vitro against DDP-resistant nasopharyngeal cancer cells and in vivo in our xenograft murine model.

The influence of anatomic location on outcomes in patients with localized primary soft tissue sarcoma.

BACKGROUND: We hypothesized that survival varied significantly between retroperitoneal soft tissue sarcoma (STS) and extremity/trunk STS. This study explored the reasons for the different outcomes and identified patient characteristics for survival. METHODS: This retrospective study identified 213 consecutive patients with localized primary STS from January 2002 to July 2013, including 47 retroperitoneal STS (22.1%) and 166 extremity/trunk STS (77.9%). Local failure-free survival (LFFS), distant metastasis-free survival (DMFS) and overall survival (OS) were constructed by the Kaplan-Meier method. Univariate Cox proportional hazards regression models were fit to assess the ability of patient characteristics to predict survival. RESULTS: At presentation, patients with retroperitoneal STS had larger tumor size (median size 18 cm vs. 6 cm; P < 0.001) and positive margin (21.3% vs. 8.4%; P = 0.014), and less often received radiotherapy (2.1% vs. 45.8%; P < 0.001). The median follow-up time for the entire population was 68 months (range, 5-127 months). Local recurrence was more frequent in patients with retroperitoneal STS compared with patients with extremity/trunk STS (53.2% vs. 28.3%; P = 0.001). LFFS and OS were lower in patients with retroperitoneal STS than extremity/trunk STS, with 5-year LFFS (50% vs. 74.3%; P < 0.001) and 5-year OS (65.4% vs. 77.5%; P = 0.017), respectively. CONCLUSION: Retroperitoneal STS was associated with significantly worse survival compared with extremity/trunk STS. Larger tumor size, more patients with positive margin and fewer patients received radiotherapy in retroperitoneal group may result in worse survival compared with extremity/trunk disease.

Size-dependent and environment-mediated shifts in leaf traits of a deciduous tree species in a subtropical forest.

AIMS: Understanding the joint effects of plant development and environment on shifts of intraspecific leaf traits will advance the understandings of the causes of intraspecific trait variation. We address this question by focusing on a widespread species Clausena dunniana in a subtropical broad-leaved forest. METHODS: We sampled 262 individuals of C. dunniana at two major topographic habitat types, the slope and hilltop, within the karst forests in Maolan Nature Reserve in southwestern China. We measured individual plant level leaf traits (i.e., specific leaf area (SLA), leaf area, leaf dry-matter content (LDMC), and leaf thickness) that are associated with plant resource-use strategies. We adopted a linear mixed-effects model in which the plant size (i.e., the first principal component of plant basal diameter and plant height) and environmental factors (i.e., topographic habitat, canopy height, and rock-bareness) were used as independent variables, to estimate their influences on the shifts of leaf traits. KEY RESULTS: We found that (1) plant size and the environmental factors independently drove the intraspecific leaf trait shifts of C. dunniana, of which plant size explained less variances than environmental factors. (2) With increasing plant size, C. dunniana individuals had increasingly smaller SLA but larger sized leaves. (3) The most influential environmental factor was topographic habitat; it drove the shifts of all the four traits examined. Clausena dunniana individuals on hilltops had leaf traits representing more conservative resource-use strategies (e.g., smaller SLA, higher LDMC) than individuals on slopes. On top of that, local-scale environmental factors further modified leaf trait shifts. CONCLUSIONS: Plant size and environment independently shaped the variations in intraspecific leaf traits of C. dunniana in the subtropical karst forest of Maolan. Compared with plant size, the environment played a more critical role in shaping intraspecific leaf trait variations, and potentially also the underlying individual-level plant resource-use strategies.

AZD3759 enhances radiation effects in non-small-cell lung cancer by a synergistic blockade of epidermal growth factor receptor and Janus kinase-1.

AZD3759 is a novel epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI) on the basis of gefitinib and has been proven to enter the central nervous system. Although the promising antitumor effects of AZD3759 on non-small cell lung cancer (NSCLC) have been demonstrated in clinical trials, the regulatory effects of this inhibitor on the antitumor efficacy of radiation (RA) are unclear. The present study aimed to compare the effects of AZD3759 and osimertinib on RA efficacy in NSCLC and explore the potential mechanism of action of AZD3759. We found that the survival in RA-treated NSCLC cells was significantly decreased by treatment with 500 nM AZD3759 and osimertinib at the RA dosage of 8 Gy. The apoptotic rate, cell cycle arrest, and DNA damage in RA-treated NSCLC cells and brain metastasis in RA-treated xenograft nude mice were significantly enhanced by the co-administration of AZD3759 and osimertinib, respectively. In addition, AZD3759 showed a significantly stronger efficacy than osimertinib did. Mechanistically, the receptor tyrosine kinase signaling antibody array revealed that Janus kinase-1 (JAK1) was specifically inhibited by AZD3759, but not by osimertinib. The effects of AZD3759 on RA efficacy in PC-9 cells and in a brain metastasis animal model were significantly abolished by the overexpression of JAK1. Collectively, our results suggested that AZD3759 promoted RA antitumor effects in NSCLC by synergistic blockade of EGFR and JAK1.

AZD3759 inhibits glioma through the blockade of the epidermal growth factor receptor and Janus kinase pathways.

Glioma is an intracranial malignant tumor with high morbidity in China. Limited efficacy has been achieved in the treatment of glioma through the application of epidermal growth factor receptor (EGFR) inhibitors, which is reported to be related to the poor permeability of the brain-blood barrier (BBB) to EGFR inhibitors. AZD3759 and osimertinib are both BBB-penetrating EGFR inhibitors. The present study aimed to investigate the inhibitory effects of AZD3759 and osimertinib on glioma and compare their efficacy and the underlying mechanisms. C6 and U87 cells were incubated with different concentrations of AZD3759 (1, 2, and 4 µM) and 4 µM osimertinib, respectively. C6-LUC xenograft animals were administered different doses of AZD3759 (15, 30, and 60 mg/kg) and 60 mg/kg osimertinib. We found that proliferation was significantly suppressed and that apoptosis and cell cycle arrest were dramatically induced in both C6 and U87 cells by AZD3759 in a dose-dependent manner. Compared to AZD3759, osimertinib had inferior effects on proliferation, apoptosis, and cell cycle. In vivo experiments verified that the anti-tumor efficacy of AZD3759 against C6 xenograft tumors was dose dependent and superior to that of osimertinib. The inhibitory effects of AZD3759 on the Janus kinase (JAK)/STAT pathway were observed in both glioma cells and tumor tissues, which were more significant than those of osimertinib. In conclusion, AZD3759 may inhibit the progression of glioma via a synergistic blockade of the EGFR and JAK/STAT signaling pathways.

The survival benefit of radiotherapy in localized primary adult rhabdomyosarcoma.

AIM: To evaluate the role of radiotherapy (RT) in the treatment of localized primary adult rhabdomyosarcoma (RMS). METHODS: This retrospective study identified 62 consecutive adult patients with localized primary RMS from January 2000 and July 2016. Local failure-free survival (LFFS), distant metastasis-free survival (DMFS) and overall survival (OS) were analyzed by the Kaplan-Meier method. Multivariate Cox proportional hazards regression models were fit to assess the ability of patient characteristics to predict survival. RESULTS: With a median follow-up of 33 months (range, 6-195 months), the 5-year LFFS, DMFS and OS of all patients were 64.0%, 50.0% and 45.0%, respectively. RT was administered to 28 patients (45.2%). Patients who received RT had a higher 5-year LFFS (81.7% vs 47.2%), 5-year DMFS (59.4% vs 43.1%) and 5-year OS (57.1% vs 34.8%) compared with patients who did not received RT. In mulitvariate analysis, RT retained significance as an independent predictor of improved LFFS [hazard ratio (HR) = 0.282; 95% confidence interval (CI), 0.095-0.838; P = 0.023], DMFS (HR = 0.289; 95% CI, 0.125-0.991; P = 0.004) and OS (HR = 0.334; 95% CI, 0.153-0.727; P = 0.006). CONCLUSIONS: RT significantly reduced local recurrence, distant metastasis and tumor mortality compared with no radiotherapy for localized primary adult RMS.

MicroRNA-449b-5p suppresses cell proliferation, migration and invasion by targeting TPD52 in nasopharyngeal carcinoma.

Nasopharyngeal carcinoma (NPC) is an important type of head and neck malignant cancer with geographical distribution. MicroRNA-449b-5p (miR-449b-5p) is related to the development of various cancers, while its function in NPC remains unknown. The present study aimed to investigate the role and target gene of miR-449b-5p in NPC. Expressions of miR-449b-5p in NPC cell lines and clinical tissues were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Cell proliferation was determined by MTT and colony formation assays. Migration and invasion abilities after different treatment were evaluated by wound healing and Transwell assays, respectively. Dual-luciferase reporter assay was performed to explore the relationship between miR-449b-5p and tumour protein D52 (TPD52). TPD52 expression was determined by qRT-PCR and western blot assay. miR-449b-5p was significantly downregulated in NPC cell lines and clinical tissues than the matched control. Overexpression of miR-449b-5p inhibited proliferation, migration and invasion of NPC cells. Dual-luciferase reporter assay indicated that miR-449b-5p directly targeted TPD52. Furthermore, shRNA-mediated downregulation of TPD52 rectified the promotion of cell migration and invasion by miR-449b-5p inhibition. In conclusion, the present study suggests that miR-449b-5p, as a novel tumour-suppressive miRNA against NPC, inhibits proliferation, migration and invasion of NPC cells via inhibiting TPD52 expression.

MiR-194 regulates nasopharyngeal carcinoma progression by modulating MAP3K3 expression.

Despite the recent development of treatment strategies for nasopharyngeal carcinoma, the effective management of this disease remains a challenging clinical problem. A better understanding of the regulatory roles of miR-194 and mitogen-activated protein kinase kinase kinase 3 (MAP3K3) in the nasopharyngeal-carcinoma-related gene network is required to address this issue. Here, we measured relative expression of miR-194 in human nasopharyngeal carcinoma tissues and normal epithelial tissues by quantitative real time PCR. We transfected cultured CNE-1 and C666-1 cells with miR-194 mimics, and then examined the effects on cell proliferation, cell migration and invasion. Luciferase reporter assay was used to validate the putative binding between miR-194 and MAP3K3. We then examined the effect of knockdown and overexpression of MAP3K3 on cell tumorigenesis. Expression of miR-194 is significantly down-regulated in nasopharyngeal carcinoma specimens and tumor cell lines when compared with normal controls. In addition, miR-194 suppressed tumor cell proliferation and viability, as well as migration and invasion of carcinoma cells. We found that miR-194 binds the 3' untranslated region of MAP3K3, and knockdown of miR-194 inhibited nasopharyngeal carcinoma cell proliferation, migration and invasion. In accordance, overexpression of MAP3K3 reversed the inhibitory effects of miR-194 in carcinoma cells. This study suggests that expression of miR-194 is down-regulated in nasopharyngeal carcinoma, and that miR-194 can directly target MAP3K3 to regulate tumor progression. Given the pivotal involvement of MAP3K3 in nasopharyngeal carcinoma development, targeting miR-194 may be a novel strategy for the treatment of nasopharyngeal carcinoma.

ALK and ROS1 concurrent with EGFR mutation in patients with lung adenocarcinoma.

PURPOSE: The purpose of this study was to explore the frequencies of ALK and ROS1 fusion genes in EGFR-mutant lung adenocarcinoma patients and examine the therapeutic efficacies of EGFR-tyrosine kinase inhibitors (TKIs). MATERIALS AND METHODS: A total of 421 EGFR-mutated patients taking EGFR-TKIs were examined for ALK and ROS1 fusion genes based on reverse transcription-polymerase chain reaction (RT-PCR). Progression-free survival (PFS) and overall survival (OS) were evaluated by the Kaplan-Meier method and compared by the log-rank test. RESULTS: The mutations of ALK rearrangement (n=10) and ROS1 rearrangement (n=3) were detected. All the patients received EGFR-TKIs, and eight took subsequent ALK/ROS1 inhibitor. PFS was longer in single EGFR mutants (n=408) than in EGFR/ALK or EGFR/ROS1 counterparts (n=13; 10.7 vs 6.6 months, P=0.004). No difference in OS existed between single EGFR and EGFR/ALK or EGFR/ROS1 mutants (21.0 vs 23.0 months, P=0.196). The median PFS of eight patients treated with ALK/ROS1 inhibitor was 6.0 months. CONCLUSION: Concomitant ALK/ROS1 fusion genes occurred in 3.1% EGFR-mutated lung adenocarcinoma patients. Concomitant ALK/ROS1-EGFR mutations may influence the therapeutic efficacy of EGFR-TKIs.

MiR-205 promotes proliferation, migration and invasion of nasopharyngeal carcinoma cells by activation of AKT signalling.

OBJECTIVE: To examine the role of microRNA (miR)-205 in proliferation, migration and invasion of nasopharyngeal carcinoma (NPC). METHODS: The human NPC cell line CNE2 was transfected with miR-205 mimic, anti-miR-205 inhibitor or scrambled oligonucleotide (control). Cell proliferation was assessed via MTT assay. Cell migration and invasion were evaluated by transwell migration and Matrigel® invasion assay, respectively. Radiation induced apoptosis was quantified via Caspase-Glo3/7 assay. Apoptotic proteins and epithelial-mesenchymal transition (EMT) proteins were semiquantified by Western blot analysis. RESULTS: Overexpression of miR-205 increased the proliferation, migration and invasion of CNE2 cells, and decreased radiation-induced apoptosis compared with control cells. MiR-205 overexpression downregulated E-cadherin and upregulated Snail expression via downregulation of PTEN and upregulation of AKT. CONCLUSION: MiR-205 plays vital roles in tumourigenesis and tumour progression in NPC, and may be a potential treatment target.

Treatment and survival analyses of completely resected thymic carcinoma patients.

PURPOSE: To investigate the impact of chemotherapy and/or radiotherapy on disease-free survival (DFS) and overall survival (OS) rates of patients with thymic carcinoma after complete resection. METHODS: Between 2001 and 2013, 54 patients with complete resection of thymic carcinoma in Hangzhou Cancer Hospital were retrospectively reviewed. The Kaplan-Meier method was used to evaluate the survival rates. The Cox proportional hazard model was used for multivariate analysis. RESULTS: Among the 54 patients, Masaoka stage I was observed in seven patients, II in 22 patients, and III in 25 patients. Sixteen patients received adjuvant chemotherapy (six with chemotherapy alone and ten with radiotherapy and chemotherapy), 25 patients received adjuvant radiotherapy, and 13 patients did not receive radiotherapy and/or chemotherapy. The 5-year DFS and OS rates for all patients were 63.0% and 73.4%, respectively. Univariate analysis revealed that radiotherapy was significantly associated with DFS and OS (P=0.014 and P=0.029, respectively), while adjuvant chemotherapy was not (P=0.122 and P=0.373, respectively). Multivariate analysis showed that adjuvant radiotherapy increased DFS (P=0.041), but not OS (P=0.051). CONCLUSION: Complete resection followed by adjuvant radiotherapy increased disease-free rates of thymic carcinoma patients.

[Clinical study of acute low-tone sensorineural hearing loss].

OBJECTIVE: To study the etiology, diagnosis and therapeutic strategies of acute low-tone sensorineural hearing loss (ALHL). METHOD: Fifty-five patients (58 ears) with ALHL were selected for this study. With detailed history collection, otological examination and pure tone audiometry,acoustic immittance and auditory brainstem response (ABR), all cases were divided into two groups, the prednisone group and the control group, randomly with six months follow-up. RESULT: ALHL mainly affected young people. The symptoms included the sensation of ear fullness, low-tone tinnitus, and hearing impairment. Otological examinations showed normal results. Mild to moderate sensorineural hearing loss at low frequencies and type "A" tympanograms were found in all cases. Acoustic stapedial reflexes were elicited in 49 out of 58 affected ears and 40 of them had positive results on the Metz test. ABR responses were normal in all 45 tested ears. with comparison of the two groups, the effect of prednisone group was superior to that of the control group. CONCLUSION: ALHL patients are clinically characterized by low-tone tinnitus, aural fullness and hearing loss, which mainly involve unilateral ear. Audiological findings indicate cochlear impairment, which only invades low frequency region. This study suggest that the corticosteroid therapy probably have some effect.