The level of serum carcinoembryonic antigen is a surrogate marker for the efficacy of EGFR-TKIs but is not an indication of acquired resistance to EGFR-TKIs in NSCLC patients with EGFR mutationsm.

The aim of the present study was to define the relationship between carcinoembryonic antigen (CEA) and survival in non-small cell lung cancer (NSCLC) patients receiving epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) and to investigate whether the level of serum CEA is related to the mechanism for acquisition of resistance to EGFR-TKIs. A total of 100 patients with advanced NSCLC (stage IIIB or stage IV) and harboring EGFR mutations were included. All patients received erlotinib or gefitinib treatment. The correlation between CEA serum level and clinical benefit from erlotinib or gefitinib treatment was analyzed. Patients were appraised by a review of data from a prospective re-biopsy protocol for lung cancer patients with an EGFR-mutated phenotype with acquired resistance to EGFR-TKI therapy. Of 100 patients, 49 and 21 patients carried high and low level of CEA, respectively; 30 carried normal CEA. Median progression-free survival was 6.4 and 4.5 months in patients with high and low level of CEA, respectively (P=0.027). Median PFS of patients in low-CEA group longer than that of those with normal level of tumors (3.0 months; P=0.002). The difference between groups L and N was not significant regarding objective response rate and overall survival. No significant difference was found in three groups of acquired resistance to EGFR-TKIs. The relative CEA level could predict benefit of EGFR-TKI therapy in advanced NSCLC, but could not predict acquired resistance to EGFR-TKIs.

Fast-track surgery versus conventional perioperative management of lung cancer-associated pneumonectomy: a randomized controlled clinical trial.

BACKGROUND: The aim of this study is to investigate the effects of fast-track surgery (FTS) on postoperative recovery, hospital stay, total medical costs, and the complications of pneumonectomy in patients with non-small cell lung cancer (NSCLC). METHODS: Studies were performed between June 2012 and March 2014 in 17 patients received FTS and 18 patients given conventional management (control) after pneumonectomy in the Department of Thoracic Surgery, the Fourth Affiliated Hospital of Harbin Medical University. Patients were evaluated based on their days to achieve the first postoperative flatus, C-reactive protein (CRP) at postoperative day (POD) 1-7, the length of hospital stay, the medical costs, and postoperative complications. RESULTS: The results showed that in the FTS group, latency to the first postoperative flatus (1.5 ± 0.6 versus 3.1 ± 0.8 s in controls, P < 0.0001), CRP (71.36 ± 5.48 versus 80.71 ± 8.32 mg/L in at POD 7, P < 0.0001), the length of hospital stay (18.1 ± 1.4 versus 27.4 ± 6.6 days, P < 0.0001), and the medical costs (29.9 ± 2.7 versus 37.2 ± 3.6 thousand Chinese Yuan, P < 0.0001) were significantly reduced compared to the group receiving conventional management. FTS group also had a relatively lower postoperative complication rate (23.5% of 17 versus 33.3% of 18 in control group) although it was statistically insignificant (P = 0.711). CONCLUSIONS: These results indicate that application of the FTS in NSCLC pneumonectomy efficiently accelerates postoperative recovery, shortens hospital stay, reduces the total medical costs of the patients and thus is more acceptable than conventional management.

Upregulation of miR-24 promotes cell proliferation by targeting NAIF1 in non-small cell lung cancer.

Recent studies have implied that aberration of miR-24 is linked to various human cancers. However, its role in non-small cell lung cancer (NSCLC) remains obscure. Here, we found that miR-24 was significantly upregulated in NSCLC tissues and patients' serum. High expression of miR-24 in patients' serum was independently correlated with a shorter overall survival of NSCLC patients. Depletion of miR-24 inhibited cell proliferation and anchorage-independent survival ability in lung cancer cell lines and reduced tumor formation ability in nude mice. Nuclear apoptosis-inducing factor 1 (NAIF1) was identified to be a functional target of miR-24 in the human lung. Next, we observed that the NAIF1 mRNA expression level in NSCLC tissues was suppressed in comparison to that in adjacent normal tissues. Restoration of NAIF1 in lung cancer cell inhibited cell proliferation and anchorage-independent survival ability, which were found to be similar with those from transfecting a miR-24 inhibitor into lung cancer cells. In conclusion, our study demonstrated that miR-24 was upregulated in NSCLC, and suppressing the expression of miR-24 inhibited tumor characteristics. MiR-24 acted as an oncomir, at least partially through regulation of its functional target NAIF1 in NSCLC. MiR-24 may serve as a novel potential biomarker for NSCLC diagnosis and prognosis.

Activation of STAT5 contributes to proliferation in U87 human glioblastoma multiforme cells.

Rapid increases in the tyrosine phosphorylation of signal transducers and activators of transcription 5 (STAT5) proteins have been extensively documented in cells stimulated with cytokines and growth factors. However, the mechanisms by which STAT5 translocates to the nucleus and regulates proliferation in human glioblastoma multiforme cells have not been studied in detail. To the best of our knowledge, the present study demonstrated for first time that stimulation of a glioblastoma multiforme (GBM) cell line (U87-MG) with hepatocyte growth factor (HGF) resulted in the phosphorylation of STAT5 at Tyr-694/699 and nuclear translocation of STAT5. In addition, HGF promoted nuclear translocation of STAT5 in a time-dependent manner and increased the proliferation of U87-MG cells. In order to determine the role of STAT5 directly, RNA interference was used to knockdown STAT5 expression in the U87-MG cell line. It was illustrated that small interfering RNA (siRNA) against STAT5 successfully inhibited the protein expression of STAT5 in the U87-MG cell line, leading to a potent suppression of tumor cell proliferation with or without HGF treatment. In order to broaden the investigation and to determine the role of STAT5 in vivo, immunohistochemistry (IHC) was applied to evaluate STAT5 expression in 100 newly diagnosed glioma and 10 non-neoplastic brain tissues. p-STAT5 expression increased according to the histopathological grade of the glioma. However, no p-STAT5 staining was observed in non-neoplastic brain tissues. These findings suggested that inhibition of the STAT5 pathway may be an effective therapeutic strategy for the clinical management of GBM.

[Effect of early enteral nutrition on immune response and clinical outcomes after esophageal cancer surgery].

OBJECTIVE: To evaluate the effect of early enteral nutrition(EEN) on immune response and clinical outcomes after esophageal cancer operation. METHODS: Sixty patients with esophageal cancer undergoing radical operation between March 2010 and July 2011 were randomly divided into two groups using envelope method: EEN group(n=30, administration of water and enteral nutrition early after operation) and TPN group(n=30, administration of total parenteral nutrition). Two groups both received 7-day nutrition support. Immune indexes(CD3(+), CD4(+), CD8(+), CD4(+)/CD8(+)) and serum nutritional indexes(albumin, pre-albumin, transferrin) were measured before operation and 1-, 3-, and 7-day after operation. The time to first flatus, length of postoperative hospital stay, total hospitalization cost, and postoperative complication were recorded. RESULTS: As compared to TPN group, the time to first flatus was significantly shorter in EEN group[(66.5±7.3) h vs. (75.1±6.8) h, P<0.01], as was hospital stay[(7.8±1.1) d vs. (9.3±1.3) d, P<0.01]. Total hospitalization cost[(36 210±3810) yuan vs. (39 731±4013) yuan, P<0.01] was lower in EEN group as compared to TPN group. There was no significant difference in postoperative complication rate between the two groups[13.3%(3/30) vs. 20.0%(6/30), P>0.05]. The levels of CD3(+), CD4(+), CD4(+)/CD8(+), albumin, prealbumin and transferrin were significantly higher in EEN group as compared to TPN group on postoperative day 3 and 7(all P<0.05), while CD8(+) was significantly lower in EEN group(P<0.05). CONCLUSION: EEN can promote early recovery of gastrointestinal function, improve nutritional and immune function, and therefore lead to fast postoperative recovery in esophageal cancer patients.

Fast-track surgery improves postoperative clinical recovery and reduces postoperative insulin resistance after esophagectomy for esophageal cancer.

PURPOSE: This study aims to investigate the influence of fast-track surgery (FTS) on insulin resistance indicators in a prospective randomized, controlled clinical trial in esophageal cancer patients. METHODS: Between November 2009 and March 2011, 34 patients underwent the FTS pathway, and 34 patients underwent the conventional pathway after esophagectomy in our department. The times to postoperative flatus and defecation, duration of postoperative hospital stay, hospitalization expenditures, and postoperative complications were recorded. Insulin resistance indicators were measured before operation as well as on the 1st, 3rd, and 7th postoperative days (PODs), including serum levels of fasting blood glucose (FBG), fasting insulin (FINS), interleukin-6 (IL-6), and C-reactive protein (CRP) in patients of both groups. The insulin resistance index (homeostasis model assessment of insulin resistance (HOMA-IR)) was calculated at each time point. RESULTS: We found a significantly shorter postoperative hospital stay and faster return of gastrointestinal function in patients who underwent FTS (P < 0.01). In addition, the total hospitalization expenditure was significantly lower in the FTS group (P < 0.01). The preoperative insulin resistance indicators showed no significant differences between the two groups. On PODs 1 and 3, the levels of log-HOMA-IR, FINS, IL-6, and CRP in the FTS group were significantly lower than those in the control group (all P < 0.05). On POD 7, the CRP level in the FTS group was significantly lower than that in the control group (P < 0.05). CONCLUSIONS: FTS promotes early recovery of gastrointestinal function and reduces stress reaction and postoperative insulin resistance after esophagectomy for esophageal cancer.

[Mechanisms of EGF regulation of COX-2 through the STAT5 signaling pathway in human lung adenocarcinoma A549 cells].

BACKGROUND: It has been proved that cyclooxygenase-2 (COX-2) is a key factor in lung cancer oncogenesis. COX-2 can be induced by a number of cytokines and growth factors and can be regulated by the JAK/STAT signaling pathway. Inhibiting the expression of COX-2 can prevent the development of lung cancer. The aim fo this study is to investigate whether the epidermal growth factor (EGF) can stimulate the signal transducers and activators of transcription 5 (STAT5) as well as to discover the effects of the STAT5 signaling pathway on the COX-2 in human lung adenocarcinoma A549 cells. METHODS: The phenomenon of STAT5 activation stimulated by the EGF was assayed through immunofluorescence and Western blot. The adenovirus containing the wild-type (WT)-STAT5 (AdWT-STAT5) plasmid, dominant-negative (DN)-STAT5 (Ad-CMV5Stat5aΔ740) plasmid, and STAT5 siRNA were transfected into A549 cells. The latter two groups were stimulated using EGF. Reverse transcriptase polymerase chain reaction was used to detect the mRNA expression of COX-2. RESULTS: STAT5 was not activated in A549 cells in vitro. EGF stimulation significantly increased the level of the p-STAT5 protein and induces the shuttling of p-STAT5 from the cytoplasm into the nucleus. STAT5 activation was crucial for the COX-2 expression induced by the EGF. STAT5 was required for COX-2 expression, but can mediated the effects of the COX-2 expression through pathways that were independent of transcriptional activation. CONCLUSIONS: COX-2 expression is dependent on STAT5 phosphorylation. A second pathway does not require STAT5 phosphorylation.

Fast-track rehabilitation program and conventional care after esophagectomy: a retrospective controlled cohort study.

PURPOSE: The purpose of this article is to evaluate fast-track rehabilitation program and conventional care after esophagectomy using a retrospective controlled cohort study in esophageal cancer patients. METHODS: Fifty-five patients underwent fast-track rehabilitation program and 57 patients underwent conventional care after esophagectomy. Fast-track rehabilitation program was performed to patients who have early movement, epidural analgesia control, fluid infusion volume control and enteral nutrition for early discharge. The other 57 patients underwent conventional care after esophagectomy. The average of hospital stay and complications were calculated in the patients between the two groups. RESULTS: The median length of hospital stay in the patients was significantly shorter after fast-track rehabilitation program than after conventional care (7.7 vs 14.8 day, P < 0.01). The percentage of patients who developed complications was significantly lower 30 day after fast-track rehabilitation program than after conventional care (29.1 vs 47.4%, P < 0.05). 87.3% in patients of the fast-track rehabilitation program group and 54.4% in those of the conventional care group reported excellent to very good satisfaction with their pain control (P = 0.000). CONCLUSIONS: The fast-track rehabilitation program results in fewer complications, less postoperative pain, a reduction in the hospital length of stay, and quicker return to work and normal activities after esophagectomy.

Fluorescence in situ hybridization as adjunct to cytology improves the diagnosis and directs estimation of prognosis of malignant pleural effusions.

BACKGROUND: The identification of malignant cells in effusions by conventional cytology is hampered by its limited sensitivity and specificity. The aim of this study was to investigate the value of fluorescence in situ hybridization (FISH) as adjuncts to conventional cytologic examination in patients with malignant pleural effusions. METHODS: We conducted a retrospective cohort study of 93 inpatients with pleural effusions (72 malignant pleural effusions metastatic from 11 different organs and 21 benign) over 23 months. All the patients came from Chinese northeast areas. Aspirated pleural fluid underwent cytologic examination and fluorescence in situ hybridization (FISH) for aneuploidy. We used FISH in single-colour or if appropriate in dual-colour evaluation to detect chromosomal aberrations (chromosomes 7, 11, and 17) in effusion cells as markers of malignancy, to raise the diagnostic yield and identified the efficiency by diagnostic biopsy. Predominant cytogenetic anomalies and patterns of intratumor cytogenetic heterogeneity were brought in relation to overall survival rate. RESULTS: Cytology alone confirmed malignant pleural effusions in 45 of 72 patients (sensitivity 63%), whereas FISH alone positively identified 48 of 72 patients (sensitivity 67%). Both tests had high specificity in predicting benign effusions. If cytology and FISH were considered together, they exhibited 88% sensitivity and 94.5% specificity in discriminating benign and malignant effusions. Combined, the two assays were more sensitive than either test alone. Although the positive predictive value of each test was 94.5%, the negative predictive value of cytology and FISH combined was 78%, better than 47% and 44% for FISH and cytology alone, respectively. There was a significantly prolonged survival rate for patients with aneuploidy for chromosome 17. CONCLUSIONS: FISH in combination with conventional cytology is a highly sensitive and specific diagnostic tool for detecting malignant cells in pleural effusions . The high sensitivity and specificity may be associated with geographic area and race. Simple numeric FISH anomalies may be prognostic.

Calcineurin/NFATc1 pathway contributes to cell proliferation in hepatocellular carcinoma.

BACKGROUND AND AIMS: The nuclear factor of the activated T cell (NFAT) family was primarily recognized for its central role in T lymphocyte activation. Recent evidence showed that NFAT isoforms participate in the regulation of genes related to cell proliferation and differentiation in epithelial malignancies. Here, we investigated the expression and activation of the calcineurin/NFAT transcription pathway and its role in hepatocellular carcinoma (HCC) proliferation. METHODS: Expression of NFATc1 and calcineurin proteins was examined by immunohistochemical analyses in 76 human HCC samples. The cellular NFAT activation and distribution in HepG2 cells were analyzed by immunofluorescence and western blot analyses. After NFATc1 expression was knocked down by NFATc1-specific siRNA, we analyzed its implications in cell cycle progression and growth by MTT and flow cytometry. The impact of calcineurin/NFAT signaling on protein expression of c-myc and cox-2 were performed by western blot analyses. RESULTS: NFATc1 is significantly overexpressed in HCC. The regulation of calcineurin activity by ionomycin or cyclosporin A caused rapid nuclear import or export of NFATc1 in HepG2 cells. NFATc1 knock-down led to a significant reduction in proliferation rates and cell cycle arrest at G1 phase. The expression of c-myc and cox-2 was decreased in the NFATc1 knock-down HepG2 cells. Ionomycin increased c-myc and cox-2 expression in HepG2 cells, but not in siNFATc1 HepG2 cells. CONCLUSION: The calcineurin/NFATc1 signal is overexpressed and active in HCC. It may enhance the proliferative potential of HepG2 cells through transcriptional activation of the c-myc and cox-2 oncogenes.

EGF stimulates cyclooxygenase-2 expression through the STAT5 signaling pathway in human lung adenocarcinoma A549 cells.

The epidermal growth factor receptor (EGFR) can be activated by several growth factors within the tumor microenvironment, and it can activate several signaling pathways. For tumor development, these EGFR-related signaling pathways may converge on several common nuclear transcription factors, one such transcription factor being STAT5. STAT5 plays an important role in the oncogenic signal transduction pathway in non-small cell lung cancer. In this study, we examined whether the epidermal growth factor (EGF) can stimulate cyclooxygenase-2 (COX-2) expression in human lung adeno-carcinoma A549 cells transfected with or without STAT5 siRNA or dominant-negative (DN)-STAT5, and identified the pathways involved in this response. We found that STAT5 siRNA significantly reduced EGF-induced COX-2 expression, and STAT5 phosphorylation. STAT5 phosphorylation predominantly mediates EGF-induced COX-2 promoter activity. STAT5 siRNA was found to inhibit COX-2 expression in resting A549 cells despite the absence of detectable activated phosphorylated STAT5. Using an adenoviral system, we expressed DN-STAT5 in human lung adenocarcinoma A549 cells in order to broaden the investigation and to determine the role of STAT5 in EGF-mediated COX-2 gene expression. The overexpression of DN-STAT5 significantly inhibited EGF-induced COX-2 expression, and we found that EGF induced the tyrosine phosphorylation of STAT5 and up- regulated COX-2 expression. DN-STAT5 also blocked COX-2 promoter activity. Our results demonstrate that EGF stimulates COX-2 expression in human lung adenocarcinoma A549 cells via the activation of the STAT5 pathway and that COX-2 expression may be independent of phosphorylated STAT5 in A549 cells in vitro.

STAT5 regulates glioma cell invasion by pathways dependent and independent of STAT5 DNA binding.

STAT5 activation in primary glioma was analyzed using antibody directed against phosphorylated STAT5 (pSTAT5), in all samples robust pSTAT5 immunostaining was detected, predominantly in the nucleus. We then used immunofluorescence, transcription factor binding assays and western blotting to study EGF-modulated STAT5 activation in the human U87 glioma cell line. EGF was found to upregulate pSTAT5 levels and enhances STAT5 DNA-binding activity. To address the role of STAT5 in glioma cell invasion, resting and EGF-treated U87 cells were treated with siRNA directed against STAT5 and the extent of glioma cell migration into a Matrigel matrix was monitored. EGF treatment markedly enhanced matrix invasion, and knockdown of STAT5 expression with siRNA significantly downregulated matrix invasion by both EGF-stimulated and resting glioma cells. However, transfection with a decoy oligonucleotide abolished transcriptional activation of a STAT5 target gene but failed to inhibit matrix invasion in resting U87 cells. By contrast, the decoy nucleotide abolished EGF enhancement of matrix invasion. STAT5 therefore promotes glioma cell invasion via at least two different pathways: a pathway dependent of STAT5 DNA binding, and a second pathway independent of STAT5 DNA binding.

Induction of cytotoxicity by photoexcitation of TiO2 can prolong survival in glioma-bearing mice.

We have investigated the possibility that photoexcited titanium dioxide (TiO2) could inhibit the growth of malignant cells. We studied the anti-glioma effects of nano-TiO2 excited with ultraviolet A (UVA) irradiation both in vitro and in vivo. Transmission electron microscopy demonstrated that glioma cells take up TiO2 by phagocytosis, and vital staining revealed that TiO2 alone has no effect on glioma cell proliferation. However, if TiO2 was combined with UVA irradiation the proliferation rate was decreased significantly compared to controls (P<0.05). RT-PCR suggested that TiO2 induction of glioma cell apoptosis is associated with changes in the expression of genes encoding Bcl-2 family members. We then investigated the in vivo antitumor effects of combined TiO2 plus UVA treatment of established glioma tumors. TiO2 plus UVA led to pronounced areas of necrosis, elevated indices of apoptosis, delayed tumor growth, and increased survival compared with the TiO2-alone control group (P<0.001). Log-rank survival analysis showed that median survival duration was prolonged (P<0.001). These findings suggest that nano-TiO2 based photodynamic therapy has potential in the treatment of glioma.

TLR9 expression in glioma tissues correlated to glioma progression and the prognosis of GBM patients.

BACKGROUND: Our study aims to evaluate the expression of TLR9 in glioma tissues, examine the association between TLR9 expression, clinicopathological variables, and glioma patient outcome, we further characterized the direct effects of TLR9 agonist CpG ODN upon the proliferation and invasion of glioma cells in vitro. METHODS: RT-PCR and immunofluorescence were used to determine the expression of TLR9 in glioma cell lines and clinical glioma samples. Tissue microarry and immunohistochemistry were applied to evaluated TLR9 expression in 292 newly diagnosed glioma and 13 non-neoplastic brain tissues. We further investigated the effect of CpG ODN on the proliferation and invasion of glioma cells in vitro with MTT assays and matrigel transwell assay respectively. RESULTS: RT-PCR showed that TLR9 expressed in all the glioma samples and glioma cell lines we examined. The tissue array analysis indicated that TLR9 expression is correlated with malignancy of glioma (p < 0.01). Multivariate Cox regression analysis revealed that TLR9 expression is an independent prognostic factor for PFS of GBM patients (P = 0.026). TLR9 agonist CpG ODN has no significant effect on glioma proliferation, but matrigel transwell analysis showed that TLR9 agonist CpG ODN can significantly enhance glioma invasion in vitro. CONCLUSIONS: Our data indicated that TLR9 expression increases according to the histopathological grade of glioma, and the TLR9 expression level is related to the PFS of GBM patients. In addition, our findings warrant caution in the directly injection of TLR9 agonist CpG ODN into glioma tissues for the glioma immunotherapy.