Identification of a 4-lncRNA signature predicting prognosis of patients with non-small cell lung cancer: a multicenter study in China.

BACKGROUND: Previous findings have indicated that the tumor, nodes, and metastases (TNM) staging system is not sufficient to accurately predict survival outcomes in patients with non-small lung carcinoma (NSCLC). Thus, this study aims to identify a long non-coding RNA (lncRNA) signature for predicting survival in patients with NSCLC and to provide additional prognostic information to TNM staging system. METHODS: Patients with NSCLC were recruited from a hospital and divided into a discovery cohort (n = 194) and validation cohort (n = 172), and detected using a custom lncRNA microarray. Another 73 NSCLC cases obtained from a different hospital (an independent validation cohort) were examined with qRT-PCR. Differentially expressed lncRNAs were determined with the Significance Analysis of Microarrays program, from which lncRNAs associated with survival were identified using Cox regression in the discovery cohort. These prognostic lncRNAs were employed to construct a prognostic signature with a risk-score method. Then, the utility of the prognostic signature was confirmed using the validation cohort and the independent cohort. RESULTS: In the discovery cohort, we identified 305 lncRNAs that were differentially expressed between the NSCLC tissues and matched, adjacent normal lung tissues, of which 15 are associated with survival; a 4-lncRNA prognostic signature was identified from the 15 survival lncRNAs, which was significantly correlated with survivals of NSCLC patients. This signature was further validated in the validation cohort and independent validation cohort. Moreover, multivariate Cox analysis demonstrates that the 4-lncRNA signature is an independent survival predictor. Then we established a new risk-score model by combining 4-lncRNA signature and TNM staging stage. The receiver operating characteristics (ROC) curve indicates that the prognostic value of the combined model is significantly higher than that of the TNM stage alone, in all the cohorts. CONCLUSIONS: In this study, we identified a 4-lncRNA signature that may be a powerful prognosis biomarker and can provide additional survival information to the TNM staging system.

Do male esophageal cancer patients have impaired sexual function after esophagectomy? A self-reported outcomes study.

PURPOSE: Sexual function is a significant part of patients' quality of life, which is another important aspect of cancer. This study assessed and compared the sexual function of male esophageal cancer patients to that of age-matched normal controls through postoperative follow-up surveys. METHODS: The study included 105 male esophageal cancer patients aged 38-81 years who underwent a curative-intent esophagectomy between April 2012 and May 2014. This observational study included sociodemographic and clinicopathological characteristics and responses to sexual function questionnaires International Index of Erectile Function (IIEF) at 6 and 12 months after surgery. An age-matched normal control group was recruited. Non-parametric tests were used when appropriate. RESULTS: The median patient age was 59 years. The factors significantly associated with sexual dysfunction on the 6-month survey included older age, and postoperative complications. At 12 months after surgery, older age was significantly associated with poorer sexual function. The sexual function scores significantly increased from 6 to 12 months after surgery (P < 0.05); there was no difference in the patients' 12-month sexual function scores and those of the normal controls (P > 0.05). Notably, compared to older patients (age ≥60 years), the younger (age <60 years) patients reported a significantly better sexual function scores (P < 0.05). CONCLUSIONS: Age, and postoperative complications were the factors significantly associated with sexual function. Impaired sexual function after primary treatment can be recovered in male esophageal cancer patients; younger patients may regain sexual function better than their older counterparts.

Cancerous inhibitor of PP2A is targeted by natural compound celastrol for degradation in non-small-cell lung cancer.

Celastrol binds CIP2A and enhances CIP2A-CHIP interaction, leading to ubiquitination/degradation of CIP2A and inhibition of lung cancer cells in vitro and in vivo. Celastrol potentiates cisplatin's efficacy by suppressing the CIP2A-Akt pathway, and therefore CIP2A inhibitors may represent novel therapeutics for cancer.

The anticoagulant effect of PGI2S and tPA in transgenic umbilical vein endothelial cells is linked to up-regulation of PKA and PKC.

The selection of vascular grafts for coronary artery bypass surgery is crucial for a positive outcome. This study aimed to establish a novel line of vascular endothelial cells with a potent anticoagulant effect. A lentiviral vector was used to stably transfect human umbilical vein endothelial cells (HUVECs) with PGI2S alone (HUVEC-PGI2S) or both PGI2S and tPA (HUVEC-PGI2S-tPA). Both HUVEC-PGI2S and HUVEC-PGI2S-tPA cells over-expressing PGI2S and tPA were compared to mock-transfected cells. The enzyme-linked immuno sorbent assay (ELISAs) demonstrated that the anticoagulation components, ATIII and PLG, were up-regulated and coagulation factor FVIII was down-regulated in both cell lines. QRT-PCR and western blotting demonstrated the vasodilation and platelet disaggregation proteins PKA, PKC, and PTGIR were up-regulated in both cell lines, but MAPK expression was not altered in either cell line. However, cell viability and colony formation assays and cell cycle analysis demonstrated that both cell lines had a lower rate of cell growth and induced G1 phase arrest. HUVEC-PGI2S and HUVEC-PGI2S-tPA cells have a potent anticoagulant effect and their use in vascular heterografts may decrease the risk of thrombosis.

Overexpression of cystatin SN positively affects survival of patients with surgically resected esophageal squamous cell carcinoma.

BACKGROUND: Cystatin SN is a secreted protein and a cysteine proteinase inhibitor. It has been considered to be a tumor marker for gastrointestinal tract cancer in several functional researches. However, the clinicopathological and prognostic significance of Cystatin SN expression in esophageal squamous cell carcinoma (ESCC) has not been elucidated. METHODS: In our study, the expression of Cystatin SN was detected in 209 surgically resected ESCC tissues and 170 peritumoral normal esophageal mucosae by immunohistochemistry. The prognostic significance of Cystatin SN expression was analysed with Kaplan-Meier plots and the Cox proportional hazards regression models. RESULTS: The results showed that the immunostaining of Cystatin SN in ESCC tissues was less intense than that in the normal control tissue (P < 0.001). Compared with patients with low tumoral Cystatin SN expression, ESCC patients with tumors high-expression Cystatin SN exhibited increased disease-free survival (DFS) and overall survival (OS) (P < 0.001 and P < 0.001, respectively). Furthermore, the expression level of Cystatin SN could further stratify the ESCC patients by survival (DFS and OS) in the stage II subgroup (P < 0.001 and P < 0.001, respectively). Multivariate analyses showed that Cystatin SN expression, N status and differentiation were independent and significant predictors of survival. CONCLUSIONS: We concluded that ESCC patients whose tumors express high levels of Cystatin SN have favourable survival compared with those patients with low Cystatin SN expression. Tumoral Cystatin SN expression may be an independent predictor of survival for patients with resectable ESCCs.

Tobacco carcinogen induces tryptophan metabolism and immune suppression via induction of indoleamine 2,3-dioxygenase 1.

Indoleamine 2,3-dioxygenase 1 (IDO1), the enzyme that catabolizes tryptophan (Trp) metabolism to promote regulatory T cells (Tregs) and suppress CD8+ T cells, is regulated by several intrinsic signaling pathways. Here, we found that tobacco smoke, a major public health concern that kills 8 million people each year worldwide, induced IDO1 in normal and malignant lung epithelial cells in vitro and in vivo. The carcinogen nicotine-derived nitrosaminoketone (NNK) was the tobacco compound that upregulated IDO1 via activation of the transcription factor c-Jun, which has a binding site for the IDO1 promoter. The NNK receptor α7 nicotinic acetylcholine receptor (α7nAChR) was required for NNK-induced c-Jun activation and IDO1 upregulation. In A/J mice, NNK reduced CD8+ T cells and increased Tregs. Clinically, smoker patients with non-small-cell lung cancer (NSCLC) exhibited high IDO1 levels and low Trp/kynurenine (Kyn) ratios. In NSCLC patients, smokers with lower IDO1 responded better to anti-PD1 antibody treatment than those with higher IDO1. These data indicate that tobacco smoke induces IDO1 to catabolize Trp metabolism and immune suppression to promote carcinogenesis, and lower IDO1 might be a potential biomarker for anti-PD1 antibodies in smoker patients, whereas IDO1-high smoker patients might benefit from IDO1 inhibitors in combination with anti-PD1 antibodies.

High expression of Y-box-binding protein-1 is associated with poor survival in resectable esophageal squamous cell carcinoma.

BACKGROUND: Y-box-binding protein-1 (YB-1) is a multifunctional protein that regulates gene expression through both transcriptional and translational mechanism. Its expression has been associated with tumor progression and poor prognosis of many cancers. However, its role and clinical significance in resectable esophageal squamous cell carcinoma (ESCC) is still scanty. The purpose of this study was to investigate the prognostic significance of YB-1 expression by immunohistochemistry in a group of patients with ESCC treated with surgical resection. METHODS: Tissue microarray that included 233 surgically resected ESCC specimens and 49 cases of adjacent normal tissues was successfully generated for immunohistochemical evaluation. The clinical/prognostic significance of YB-1 expression was statistically analyzed. Kaplan-Meier analysis was used to compare the postoperative survival between groups. RESULTS: The results showed the immunostaining of YB-1 was distributed predominantly in cytoplasm in tumor cells, which occurred in all of the 233 patients. A higher recurrence (disease-free survival) and lower survival (overall survival) of ESCC was found in patients whose tissues had increased YB-1 expression (P<.001/P=.001). Furthermore, YB-1 expression could stratify the patient survival (disease-free survival/overall survival) in stage II (P=.012/.016). The Cox proportionate hazard regression model also established that high YB-1 expression was significantly correlated with increased risk (RR=1.752) of recurrence compared with lowYB-1 expression (P=.004). CONCLUSIONS: High expression of YB-1 is associated with poor survival in resectable ESCC patients.

Factors determining the survival of nasopharyngeal carcinoma with lung metastasis alone: does combined modality treatment benefit?

BACKGROUND: Nasopharyngeal carcinoma (NPC) with lung metastasis alone has been reported as a relatively favorable prognostic group, and combined modality treatment might be indicated for selected cases. However, the prognostic factors determining survival of this group and the indication of combined therapy have not been thoroughly studied. METHODS: We retrospectively reviewed 246 patients of NPC with lung metastasis(es) alone presented at diagnosis or as the first failure after primary treatment from 1993 to 2008 in an academic tertiary hospital. Univariate and multivariate survival analyses of post-metastasis survival (PMS) and overall survival (OS) were carried out to determine the prognostic factors. RESULTS: The 3-year, 5-year, and 10-year of PMS and OS for the whole cohort were 34.3%, 17.0%, 8.6% and 67.8%, 45.4%, 18.5%, respectively. The median PMS (45.6 months vs. 23.7 months) and OS (73.7 months vs. 46.2 months) of patients treated with combined therapy was significantly longer than that of those treated with chemotherapy alone (P < 0.001). Age, disease-free interval (DFI) and treatment modality were evaluated as independent prognostic factors of OS, while only age and treatment modality retain their independent significance in PMS analysis. In stratified survival analysis, compared to chemotherapy alone, combined therapy could benefit the patients with DFI > 1 year, but not those with DFI ≤ 1 year. CONCLUSIONS: Age ≤ 45 years, DFI > 1 year, and the combined therapy were good prognostic factors for NPC patients with lung metastasis(es) alone. The combination of local therapy and the basic chemotherapy should be considered for these patients with DFI > 1 year.

Lnc-GAN1 expression is associated with good survival and suppresses tumor progression by sponging mir-26a-5p to activate PTEN signaling in non-small cell lung cancer.

BACKGROUND: Long non-coding RNAs (lncRNAs) play vital roles in the development and progression of non-small-cell lung cancer (NSCLC); however, the role of most lncRNAs in NSCLC remains unknown. This study explored the clinical significance, biological function and underlying mechanism of lnc-GAN1 in NSCLC. METHODS: With a custom lncRNA microarray we found that lnc-GAN1 is markedly downregulated in NSCLC tissues. Then lnc-GAN1 expression level was measured using qRT-PCR in NSCLC tissues and cell lines. Survival was assessed using the Kaplan-Meier method. The biological functions of lnc-GAN1 in lung cancer cells were evaluated in vitro and in vivo. RNA fluorescence in situ hybridization and subcellular localization assays revealed the subcellular distribution of lnc-GAN1 in cells. Bioinformatic analysis was adopted to predict miRNAs and signaling pathways regulated by lnc-GAN1. RNA immunoprecipitation and Dual-luciferase reporter assays were used to assess the interaction between lnc-GAN1 and miR-26a-5p in lung cancer cells. RESULTS: lnc-GAN1 is downregulated in HCC tissues and associated with larger tumor size and poor overall survival and disease-free survival; its ectopic expression suppresses cell proliferation, colony formation, and cell cycle progression and induces apoptosis in NSCLC cells; it also inhibits tumor growth in the NSCLC xenograft model. We further proved that lnc-GAN1 is localized in cytoplasm and transcribed independently from its parental gene GAN. Mechanistically, lnc-GAN1 acts as a sponge for miR-26a-5p by two seed sequences, and the two non-coding RNAs have a negative relationship in NSCLC tissues; we further prove that PTEN is a direct target of miR-26a-5p and lnc-GAN1 inhibits cell cycle signaling pathway by activating PTEN, whose expression level correlated negatively with miR-26a-5p level but positively with lnc-GAN1 level in NSCLC samples. CONCLUSIONS: Lnc-GAN1 is downregulated and associated with poor survival of NSCLC patients, and mechanistically acts as a tumor suppressor via sponging and inhibiting miR-26a-5p to upregulate PTEN. This study provides a potential prognostic biomarker and treatment target for NSCLC.

The results and prognosis of different treatment modalities for solitary metastatic lung tumor from nasopharyngeal carcinoma: a retrospective study of 105 cases.

BACKGROUND AND OBJECTIVE: Nasopharyngeal carcinoma (NPC) is known for its propensity for distant metastases. Lung metastasis is one of the most important causes of death for patients with NPC. Solitary metastatic lung tumor from NPC is a distinctive group associated with a better survival. This study was to find a more effective treatment modality and prognostic factors for the group. METHODS: Clinical data of 105 cases of solitary metastatic lung tumor from NPC were retrospectively analyzed. Survival rate was calculated by the Kaplan-Meier method. The difference of survival between the patients treated by different modalities was evaluated by the log-rank test. The Cox univariate and multivariate analyses of gender, age, pathologic type, stage, adjuvant chemotherapy, evaluation of treatment for NPC, disease-free interval, size of metastatic tumor, pulmonary hilar and/or mediastinal lymph node metastasis, treatment modalities, recurrent distant metastases and/or relapse of NPC were conducted. RESULTS: The local control rate was 53.8% in chemotherapy group, 88.0% in radiotherapy ± chemotherapy group, and 96.4% in operation ± chemotherapy group (P < 0.01). The most promising progression-free survival (PFS) and overall survival (OS) were obtained with operation ± chemotherapy and followed by radiotherapy ± chemotherapy. Both of them showed much better efficacy than chemotherapy (P < 0.001). The Cox multivariate analysis showed that recurrent distant metastases and/or relapse of NPC affected the survival (OR = 2.087, 95% CI = 1.277-3.410, P = 0.003). The T stage of NPC, size of metastatic tumor, hilar and/or mediastinal lymph node metastasis, and the treatment modality were independent prognostic factors. CONCLUSIONS: Operation ± chemotherapy and radiotherapy ± chemotherapy are better treatment of solitary metastatic lung tumor from NPC, which could improve the local control and prolong the PFS and OS. Chemotherapy is recommended for patients with higher T stage of NPC, size of metastatic tumor ≥ 3 cm, pulmonary hilar and/or mediastinal lymph node metastasis.

Survival analysis of 220 patients with completely resected stage-II non-small cell lung cancer.

BACKGROUND AND OBJECTIVE: Surgery is the main therapy for patients with stage II non small cell lung cancer (NSCLC), but patients still have an unsatisfactory prognosis even though complete resection is usually possible. Adjuvant chemotherapy provides low rates of clinical benefit as well. We retrospectively analyzed prognostic factors of patients with completely resected stage II NSCLC to find patients with unfavorable factors for proper management. METHODS: Clinical data of 220 patients with complete resections of stage II NSCLC at the Sun Yat sen University Cancer Center between January 1998 and December 2004 were retrospectively analyzed. Cumulative survival was analyzed by the Kaplan Meier method and compared by log rank test. Prognosis was analyzed by the Cox proportional hazards model. RESULTS: The overall 3 and 5 year survival rates were 58.8% and 47.9%, respectively. The 3 and 5 year disease free survival rates were 45.8% and 37.0%, respectively. Of the 220 patients, 86 (39.1%) had recurrence or metastasis. A univariate analysis demonstrated that age (> 55 years), blood type, the presence of symptoms, chest pain, tumor volume (> 20 cm3), total number of removed lymph nodes (> or = 10), number of involved N1 lymph nodes (> or =3 ), total number of removed N2 lymph nodes (> 6), and the ratio of involved N1 lymph nodes (> or = 35%) were significant prognostic factors for 5 year survival. In the multivariate analysis, age (> 55 years), chest pain, tumor volume (> 20 cm3), total number of removed lymph nodes (> or = 10), and number of involved N1 lymph nodes (> or = 3) were independent prognostic factors for 5 year survival. CONCLUSIONS: For patients with completely resectable stage II NSCLC, having > 55 years, presenting chest pain, tumor volumes > 20 cm3, and > or = 3 involved N1 lymph nodes were adverse prognostic factors, and > or = 10 removed lymph nodes was a favorable one. Patients with poor prognoses might be treated by individual adjuvant therapy for better survival.

Systematic identification of CDC34 that functions to stabilize EGFR and promote lung carcinogenesis.

BACKGROUND: How the oncoprotein epidermal growth factor receptor (EGFR) evades proteolytic degradation and accumulates in non-small cell lung cancer (NSCLC) remains unclear, and ubiquitin pathway genes (UPGs) that are critical to NSCLC needs to be systematically identified. METHODS: A total of 696 UPGs (including E1, E2, E3, and deubiquitinases) were silenced by small interfering RNA (siRNA) library in NSCLC cells, the candidates were verified, and their significance was evaluated in patients with NSCLC. The effects of a candidate gene on EGFR were investigated in vitro and in vivo. FINDINGS: We report 31 candidates that are required for cell proliferation, with the E2 ubiquitin conjugase CDC34 as the most significant one. CDC34 is elevated in tumor tissues in 76 of 114 (66.7%) NSCLCs and inversely associated with prognosis, is higher in smoker patients than nonsmoker patients, and is induced by tobacco carcinogens in normal human lung epithelial cells. Forced expression of CDC34 promotes, whereas knockdown of CDC34 inhibits, NSCLC cell proliferation in vitro and in vivo. CDC34 competes with c-Cbl to bind Y1045 to inhibit polyubiquitination and degradation of EGFR. In EGFR-L858R and EGFR-T790M/Del (exon 19)-driven lung tumor growth in mouse models, knockdown of CDC34 significantly inhibits tumor formation. INTERPRETATION: These results demonstrate that an E2 enzyme is capable of competing with E3 ligase to stabilize substrates, and CDC34 represents an attractive therapeutic target for NSCLCs. FUNDING: National Key Research and Development Program of China, National Natural Science Foundation of China, and the CAMS Innovation Fund for Medical Sciences.

[Short and long-term outcomes of neoadjuvant chemotherapy for locally advanced esophageal carcinoma].

OBJECTIVE: This study was to compare the 5-year survival rate, the surgical resection rate, the post-operative complications and mortality of patients who underwent surgical resection for carcinoma of esophagus with or without neoadjuvant chemotherapy. To evaluate neoadjuvant chemotherapy in the treatment of esophageal carcinoma. METHODS: Forty-two patients with locally advanced esophageal carcinoma undergoing neoadjuvant chemotherapy and surgical resection (CS group), and 75 patients with the same phase undergoing surgical resection alone (S group) from August 2003 to March 2009 in Sun Yat-sen University Cancer Center were reviewed. The 5-year survival rate, the surgical resection rate, the post-operative complications and mortality between the two groups were analyzed. RESULTS: Forty-two patients after neoadjuvant chemotherapy, the complete response rate was 11.9%, the partial response was 47.6%, the total clinical response rate was 59.5%. The surgical resection rate of CS group and S group were 100% and 89.5% (P = 0.029). There was no statistically difference in the post-operative complications and mortality between two groups. The overall 5-year survival for CS group and S group were 31.7% and 26.4%, respectively (P = 0.266). In the subgroup analysis, the 5-year survival of patients with clinical response was significant higher than S group (P = 0.010). CONCLUSIONS: The neoadjuvant chemotherapy can improve surgical resection rate and long-term survival of esophageal carcinoma patients with clinical response without increasing the post-operative complications and mortality.

Combined prognostic value of the cancer stem cell markers CD47 and CD133 in esophageal squamous cell carcinoma.

BACKGROUND: Treatments based on the inhibition of pivotal signals of cancer stem cells (CSCs) are on a promising track. Recent studies have shown that targeting CSCs with broader immune-based therapeutic methods, for example, the anti-CD47 treatment, may serve as a more potent strategy for eliminating these intractable cells. We aimed to explore the prognostic effects of CD47/CD133 and the potential therapeutic significance of CD47 in esophageal squamous cell carcinoma (ESCC). METHODS: Immunohistochemistry was employed to identify the characteristics of CD47 and CD133 in 26 pairs of tumor tissues and adjacent non-tumor tissues and 136 ESCC tissues. Kaplan-Meier analysis and Cox proportional hazards models were built for estimating the prognostic values of CD47 and CD133 expression and their combined stemness index. Sphere formation assays were undertaken to explore the effects of CD47 inhibition on primary human ESCC CSCs. RESULTS: Results conclude that CD47 and CD133 expression is increased in tumor tissues as compared to adjacent non-tumor tissues. A positive correlation between CD47/CD133 expression and differentiation was found in 136 ESCC patients. Survival analysis indicated that patients with high CD47 or CD133 expression exhibited poor overall survival and progression-free survival (PFS). The combination of high CD47 and CD133 expression was a reliable independent prognostic factor for both OS (HR = 1.940, 95% CI = 1.399-2.690, P < 0.0001) and progression-free survival (HR = 1.883, 95% CI = 1.384-2.562, P < 0.0001). Notably, CD47+ CD133+ ESCC cells were observed to possess the characteristics of CSCs, and anti-CD47 treatment veritably eliminated the CSCs pool. CONCLUSIONS: The stemness index determined by the expression of CD47 and CD133 is a promising prognostic predictor, and CD47 is a potential therapeutic target for CSCs in ESCC patients.

Somatic Mutations and Splicing Variants of Focal Adhesion Kinase in Non-Small Cell Lung Cancer.

Background: Overexpression of focal adhesion kinase (FAK) has been reported in lung cancer, but the somatic mutations and alternative splicing variants of this nonreceptor tyrosine kinase remain to be investigated. Methods: FAK in 91 lung cancer patients was sequenced using genomic DNA and cDNA samples of tumor and paired normal lung tissues as templates, and the RNA-seq data of The Cancer Genome Atlas (TCGA) data set were assessed. The biological functions of abnormal FAK transcripts and their response to FAK inhibitors were analyzed in eight cell lines using tyrosine kinase activity assay, trypan blue exclusion assay, MTT (3-(4, 5)-dimethylthiahiazo (-z-y1)-3, 5-di-phenytetrazoliumromide) assay, and transwell assay. Results: We identified an internal tandem duplication (ITD), an A1004S point mutation, an exon 5-27 deletion (ΔE5-27) truncation variant, and four FAK6,7 splicing variants (containing exons for Boxes 6 and 7) in seven (7.7%) patients. Smokers had more FAK abnormalities than nonsmokers. In FAK-ITD, the sequence encoding the C-terminal of the FERM domain and kinase domain was duplicated in-frame and produced a protein product with elevated autophosphorylation and sensitivity to FAK inhibitors. FAK6,7 was detected in the tumor but not counterpart normal lung tissues of four (4.4%) patients. In TCGA RNA-seq data, Box 6 and/or Box 7 (Box 6/7)-containing FAK variants were positive in 42 (8.3%) of 508 lung adenocarcinomas (LUADs) and 37 (7.4%) of 501 lung squamous cell carcinomas, and smokers had higher expression of Box 6/7 (+) FAK than reformed or nonsmokers with LUAD. FAK6,7 promoted cell proliferation and migration, exhibited increased autophosphorylation, and was more sensitive to FAK inhibitor compared with wild-type FAK. Conclusions: Somatic mutations and splicing variants of FAK may have a role in lung carcinogenesis and represent potential biomarkers for FAK-targeted therapies.

[Prognostic effect of mediastinal lymph node dissection in patients with stage I non-small cell lung cancer].

OBJECTIVE: To investigate the prognostic effect of mediastinal lymph node dissection in patients with stage I non-small cell lung cancer (NSCLC). METHODS: The clinical data of 330 patients with stage I NSCLC who were treated with curative resection from January 1994 to December 2003 were reviewed retrospectively. According to the extent of mediastinal lymph node dissection and the pathology report, the patients were assigned to lung resection combined with mediastinal lymph node dissection (LND) group or with lymph node sampling (LNS) group. The Kaplan-Meier method was used for survival analysis. COX proportional hazards model was used for multivariate analysis. RESULTS: There were 233 (70.6%) male patients and 97 (29.4%) female patients. The median age was 60 years old. Ninety-eight patients were in stage IA and 233 in stage IB. One hundred and forty patents were in group LND and 190 in group LNS. The mean number of removed lymph nodes in group LND and group LNS were (13.3 +/- 4.7) and (5.2 +/- 3.0) (P < 0.01), respectively. The mean of mediastinal lymph node station sampled in group LND and group LNS were (3.7 +/- 0.9) and (1.3 +/- 1.1) (P < 0.01), respectively. The 5-year and 10-year survival rates of patients in group LND were 72.0% and 66.1%, while in group LNS were 65.9% and 43.0% (P < 0.05), respectively. Other prognostic factors included symptom, staging, visceral pleura invasion and tumor size. LND was disclosed as a favourable prognostic factor at COX multivariate analysis, together with absence of symptom at diagnosis. CONCLUSION: As compared with LNS, LND can improve survival in stage I NSCLC.

The Clinical Effect of Metformin on the Survival of Lung Cancer Patients with Diabetes: A Comprehensive Systematic Review and Meta-analysis of Retrospective Studies.

Preclinical investigations have revealed an anti-cancer effect of metformin. Several studies of metformin treatment have demonstrated the improved clinical outcomes of lung cancer patients with diabetes; however, the results have been inconsistent among studies. Our systematic review and meta-analysis aimed to summarize the up-to-date effects of metformin on diabetic lung cancer patients. A systematic search was performed for studies published. Then, these studies were evaluated for inclusion, and relevant data was extracted. The summary risk estimates for the associations of metformin treatment with overall survival (OS) and progression-free survival (PFS) were analyzed using random/fixed-effects models. Analyses stratified by histological type were also conducted. Based on the 10 studies included in our analysis, metformin treatment was found to significantly improve survival, corresponding to reductions of 23% and 47% in OS [hazard ratio (HR)=0.77, 95% confidence interval (95%CI)=0.66-0.9, p=0.001] and PFS (HR=0.53, 95%CI=0.41-0.68, p<0.001), respectively. In addition, significant improvements in the OS for non-small cell lung cancer (NSCLC) (HR=0.77, 95%CI=0.71-0.84, p=0.002) and small cell lung cancer (SCLC) (HR=0.52, 95%CI=0.29-0.91, p=0.022) were observed in association with metformin treatment in analysis stratified by histological type. This stratified analysis also revealed a significant improvement in PFS for both NSCLC (HR=0.53, 95%CI=0.39-0.71, p<0.001) and SCLC (HR=0.54, 95%CI=0.34-0.84, p=0.007). We found that metformin treatment significantly improved the OS and PFS of diabetic lung cancer patients, and our findings suggest that metformin might be an effective treatment option for diabetic patients with lung cancer.

GOLPH3 promotes cell proliferation and tumorigenicity in esophageal squamous cell carcinoma via mTOR and Wnt/ß­catenin signal activation.

The authors' previous study demonstrated that Golgi phosphoprotein 3 (GOLPH3) was significantly overexpressed in esophageal squamous cell carcinoma (ESCC), correlating with poor patient survival. In the present study, GOLPH3 stable overexpression and knockdown KYSE­140 cell lines were constructed. Cell proliferation, colony formation, cell cycle progression and tumorigenesis assays were performed. The results revealed that GOLPH3 promoted ESCC cell growth and proliferation. The effects of GOLPH3 on the mechanistic target of rapamycin (mTOR) and Wnt/ß­catenin signaling pathways were investigated using western blot analyis and dual­luciferase reporter assays, and were observed to be activated in cells with GOLPH3 overexpression. Furthermore, overexpression of GOLPH3 resulted in the downregulation of p21 protein, upregulation of cyclin D1 and increased retinoblastoma­associated protein phosphorylation, consequently leading to accelerated cell cycle progression. In addition, GOLPH3 knockdown resulted in reversed effects. The results of the current study suggest that GOLPH3 serves an important role in promoting tumorigenicity of ESCC via mTOR and Wnt/ß­catenin signaling pathway activation.

The Prognostic Value of Peripheral Benzodiazepine Receptor in Patients with Esophageal Squamous Cell Carcinoma.

Background: The peripheral benzodiazepine receptor (PBR) has previously been reported as an oncogene in prostate, breast and colorectal cancers, but its prognostic value, biological behavior and function in esophageal squamous cell carcinoma (ESCC) has not been investigated. Methods: qRT-PCR, western blotting and immunohistochemistry (IHC) were used to detect PBR expression in ESCC and matched non-cancerous tissues. Based on all of the significantly independent factors, a nomogram was established to predict the prognosis of ESCC patients. In addition, we performed comprehensive in vitro experiments to study the functions of PBR in cell growth, colony formation, and migration ability, as well as its relationship with epithelial-mesenchymal transition (EMT) related proteins in ESCC cells. Results: The mRNA and protein expression levels of PBR in ESCC were higher than those in adjacent non-tumor esophageal epithelial tissues. The IHC results demonstrated that PBR expression was an independent prognostic factor in ESCC survival, patients with higher PBR expression had a poorer survival than those with low expression, and PBR expression was significantly associated with lymphoid nodal status. Furthermore, a nomogram was established to reliably predict the probability of death in ESCC patients, with a Harrell's c-index of 0.696. In the vitro experiments, knocking down the expression of PBR inhibited proliferation, colony formation and migration of ESCC cells, and regulated EMT-associated proteins (up-regulation of E-cadherin, ZO-1 and ß-catenin and concomitant with down-regulation of Fibronectin and N-cadherin). Conclusions: PBR is an independent prognostic factor in ESCC, and it promotes ESCC progression and metastasis. Basing on PBR expression level, a nomogram is established and performs a well in predicting survival of ESCC patients.

Expression and prognostic significance of MYL9 in esophageal squamous cell carcinoma.

OBJECTIVE: Myosin light chain 9 (MYL9) is necessary for cytoskeletal dynamics and experimental metastasis, but its expression in esophageal squamous cell carcinoma (ESCC) has not been addressed. We investigated the expression pattern and clinical significance of MYL9 in patients with ESCC. METHODS: We examined MYL9 expression using quantitative real-time PCR and western blotting in NE1 immortalized esophageal epithelial cells, ESCC cell lines, and paired ESCC tissues. MYL9 protein in 136 primary ESCC tissues and other types of solid tumor was detected using immunohistochemistry. The association between MYL9 expression and clinical parameters and survival was evaluated by statistical analysis. RESULTS: MYL9 was significantly upregulated in the ESCC cell lines as compared with NE1 cells. In the paired ESCC samples, MYL9 mRNA and protein expression was not significantly different between lesion tissues and the matched adjacent noncancerous tissues. In ESCC tissue, both intratumoral and peritumoral stroma were positive for MYL9. In the 136 ESCC samples, high MYL9 expression in the tumor cells significantly correlated with histological differentiation (p = 0.028), recurrence (p = 0.01), and vital status (p < 0.01). Patients with high MYL9 expression in the tumor cells had poorer overall survival (OS) and recurrence-free survival. Multivariate analysis revealed that high MYL9 expression in tumor cells was an independent and significant risk factor affecting OS after curative treatment (hazard ratio = 2.254, 95% confidence interval = 1.347-3.771, p = 0.002). CONCLUSIONS: MYL9 expression might be a promising prognostic marker and therapeutic target in ESCC.