A mutational signature associated with alcohol consumption and prognostically significantly mutated driver genes in esophageal squamous cell carcinoma.

Background: Esophageal squamous cell carcinoma (ESCC) is often diagnosed at an advanced and incurable stage. Information on driver genes and prognosticators in ESCC remains incomplete. The objective was to elucidate significantly mutated genes (SMGs), mutational signatures, and prognosticators in ESCC. Patients and methods: Three MutSig algorithms (i.e. MutSigCV, MutSigCL and MutSigFN) and '20/20+' ratio-metric were employed to identify SMGs. Nonnegative matrix factorization was used to decipher mutational signatures. Kaplan-Meier survival analysis, multivariate Cox and logistic regression models were applied to analyze association between mutational features and clinical parameters. Results: We identified 26 SMGs, including 8 novel (NAV3, TENM3, PTCH1, TGFBR2, RIPK4, PBRM1, USP8 and BAP1) and 18 that have been previously reported. Three mutational signatures were identified to be prevalent in ESCC including clocklike C>T at CpG, APOBEC overactive C>T at TpCp[A/T], and a signature featured by T>C substitution. The T>C mutational signature was significantly correlated with alcohol consumption (OR: 3.59; 95% CI: 2.30-5.67; P < 0.001). This alcohol consumption signature was also observed in liver cancer and head and neck squamous cell carcinoma, and its mutational activity was substantially higher in samples with mutations in TP53. Survival analysis revealed that TENM3 mutations (HR: 5.54; CI: 2.68-11.45; P < 0.001) and TP53 hotspot mutation p.R213* (HR: 3.37; CI: 1.73-8.06; P < 0.001) were significantly associated with shortened survival outcome. The association remained statistically significant after controlling for age, gender, TNM stage and tumor grade. Conclusions: We have uncovered several new SMGs in ESCC and defined an alcohol consumption related mutational signature. TENM3 mutations and the TP53 hotspot mutation p.R213* are independent prognosticators for poor survival in ESCC.

[A preliminary study on the relationship between laryngopharyngeal reflux and chronic rhinosinusitis].

Objective:To study the relationship between laryngopharyngeal reflux and chronic rhinosinusitis. Method:A total of 46 patients were enrolled in this study including 25 cases with chronic rhinosinusitis with nasal polyps, 10 cases with chronic rhinosinusitis without nasal polyps and 11 cases underwent surgery due to abnormal nasal anatomy such as nasal septum deviation, bubble in the turbinate, etc. as control group. The expression of pepsin was detected using immunohistochemistry in three groups. The intensity of pepsin expression and CT score of sinus, blood eosinophils percentage, blood neutrophils percentage, blood basohils percentage, blood mononuclear percentage, blood lymphocytes percentage were analyzed. Result:There were 8 strong positive cases (32%, 8/25), positive in 8 cases (32%, 8/25), 2 weakly positive cases (8%,2/25), 7 negative cases (28%, 7/25) in chronic rhinosinusitis with nasal polyps group. In the chronic rhinosinusitis without nasal polyps group, the expression of pepsin was strong positive in 4 cases (40%, 4/10), positive in 3 cases (30%, 3/10), weakly positive in 1 cases (10%, 1/10), negative in 2 cases (20%, 2/10). There were no strong positive expression in the control group, positive in 2 cases (18.2%, 2/11), weakly positive in 3 cases (27.3%, 3/11), negative in 6 cases (54.5%, 6/11), chronic rhinosinusitis with nasal polyps group and chronic rhinosinusitis without nasal polyps group higher than the control group (P<0.05). There was no significant difference in pepsin expression between chronic rhinosinusitis with nasal polyps group and without group (P=0.617). Spearman correlation analysis indicated that the intensity of pepsin was positively correlated with the score of Lund-Markay (r=0.349,P=0.017), there was no correlation with the percentage of various inflammatory cells. Conclusion:The positive expression intensity of pepsin in chronic rhinosinusitis without nasal polyps and chronic rhinosinusitis with nasal polyps is significantly higher than that in normal control group, suggested that there is a correlation between laryngopharyngeal reflux and chronic rhinosinusitis. Laryngopharyngeal reflux is positively correlated with the severity of nasal polyps. Chronic nasal inflammation caused by laryngopharyngeal reflux is not mediated by a certain kind of inflammatory cells.

Up-regulation of Twist induces angiogenesis and correlates with metastasis in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a highly invasive tumor characterized by vigorous neovascularization. The purpose of this study is to examine the expression of Twist, a highly conserved bHLH transcription factor that is known to promote EMT, and evaluate its effect on tumor angiogenesis and metastasis of HCC. The mRNA expression of Twist, VEGF, E-cadherin, and N-cadherin was determined by Real-Time RT-PCR in 30 pairs of hepatocellular carcinomas and matched non-cancerous tissues. Immunohistochemistry was carried out to analyze the protein expression of Twist, VEGF, E-cadherin, and N-cadherin in 40 hepatocellular carcinoma cases. The staining of endothelial cells for CD34 was used to evaluate the MVD. We found that Twist mRNA and protein were both increased in HCC as compared to non-cancerous tissues. The HCC specimens showing positive Twist expression had a higher microvessel density than those without Twist expression. And up-regulated Twist protein was significantly associated with intrahepatic and extrahepatic metastasis (p=0.048 and P=0.039 respectively). In addition, patients with Twist expression had poor prognosis. We also found that the expression of Twist positively correlated with up-regulation of VEGF and N-cadherin (P=0.002 and p=0.016 respectively), but not with downregulation of E-cadherin in HCC. Our results demonstrate that Twist may play an important role in the angiogenesis and metastasis of HCC. Twist expression may become a potential novel prognostic factor for the disease survival of HCC.

Reversing adriamycin resistance of human breast cancer cells by hyperthermia combined with Interferon alpha and Verapamil.

One of the major obstacles related to chemotherapy is resistance against anticancer drugs, including Adriamycin (ADM). The purpose of the present work is to investigate the reversal effects on ADM resistance by hyperthermia (42.5 degrees C) combined with two reversal agents (Interferon alpha and Verapamil) in MCF-7/ADR (ADM-resistant MCF-7 breast cancer cell line), and its relevant molecular mechanism of action. The cell survival rate and ADM IC50 of different experiment groups were measured by MTT test. The quantitative expression of MDR1 gene in cells was detected by Real-time PCR, and the expression of P-glycoprotein (P-gp) on the cells surface and the intracellular ADM accumulation was detected by flow cytometry (FCM). The ADM IC50 of the MCF-7/ADR cells decreased 830-fold after combined with Interferon alpha (IFN-alpha) and Verapamil (VRP). Although there was no distinction in the mRNA expression of MDR1, the P-gp on the MCF-7/ADR cell membrane was significantly reduced and the cellular ADM uptake increased markedly as compared to pretreatment. Our results suggeste that hyperthermia induces a considerably reversal activity against ADM resistance synergizing other reversal agents (IFN-alpha and VRP). The reversal mechanism needs further study. However, these features of hyperthermia may be exploited in clinical cancer chemotherapy.