PTK7 expression is associated with lymph node metastasis, ALK and EGFR mutations in lung adenocarcinomas.

Non-small cell lung cancer (NSCLC) is one of the leading causes of cancer death worldwide. Lung adenocarcinoma is the main tumor type of NSCLC. Recent advances in the molecular characterization and personalized therapies have improved NSCLC patient prognosis. Previous studies showed that protein tyrosine kinase 7 (PTK7) plays an important role in human cancers. However, the role of PTK7 has not been investigated. PTK7 expression was assessed by immunohistochemistry in 95 patients with lung adenocarcinoma. Correlations of PTK7 expression levels with clinicopathological parameters, EGFR mutation and EML4-ALK fusion were examined. Positive PTK7 expression was detected in 47.4% of lung adenocarcinoma. PTK7 expression was associated with gender (P=0.024), lymph node metastasis (P<0.001), ALK mutation (P=0.050), and EGFR mutations (P=0.014). No significant association was found between PTK7 expression and age (P=0.831), differentiation (P=0.494), adenocarcinoma subtype (P=0.098) and Ki67 (P=0.473). Our data suggest that PTK7 plays an oncogenic role in lung adenocarcinoma and may be a molecular marker for lymph node metastasis.

Poly (ADP-ribose) polymerase-1 Binding Protein Facilitates Lung Adenocarcinoma Cell Proliferation and Correlates with Poor Prognosis.

OBJECTIVE: This study makes a preliminary inquiry on the biological functions of PARPBP in lung adenocarcinoma progression. METHODS: mRNA expression profiles and clinical data for lung adenocarcinoma and normal lung samples were downloaded from the TCGA database. The expression of PARPBP was analyzed by a student's t test. The survival curve of lung adenocarcinoma patients was drawn using Kaplan-Meier method. A Chi-square test was applied for analyzing the correlation between PARPBP expression and clinical factors. The prognosis value of PARPBP was determined using a cox proportional hazards model. A549 cells with silenced PARPBP were constructed using RNA interference. CCK8 assay and scratch tests were conducted to test the influence of PARPBP on A549 cell proliferation and migration. A Western blot was implemented to examine the changes of p-MEK/MEK and p-ERK/ERK ratios after depleting PARPBP. RESULTS: PARPBP expression is enhanced in lung adenocarcinoma tissues (p=3.51E-35) and correlates with poor prognosis in lung adenocarcinoma patients (p=0.003). High PARPBP expression is closely correlated with pathologic-stages (p=0.033) and can be utilized as an independent predictor in lung adenocarcinoma patients. Moreover, the knockdown of PARPBP significantly suppressed A549 cell viability and migration (p<0.01). In addition, the ratios of p-MEK/MEK and p-ERK/ERK declined after silencing PARPBP (p<0.01). CONCLUSIONS: PARPBP expression is enhanced in lung adenocarcinoma tissues and is a potential factor in the progression of lung adenocarcinoma.

Aberrant Cyclin E and Hepatocyte Growth Factor Expression, Microvascular Density, and Micro-Lymphatic Vessel Density in Esophageal Squamous Cell Carcinoma.

Cyclin E and hepatocyte growth factor (HGF) have been observed as a multifaceted factor in many cancers, and the assessment of microvascular density (MVD) and micro-lymphatic vessel density (MLVD) has been used to quantify tumor angiogenesis and lymphangiogenesis. The aim of this study was to explore the association between expression of cyclin E, HGF, MVD, and MLVD, and clinicopathologic parameters in esophageal squamous cell carcinoma (ESCC). The expression of cyclin E, HGF, MVD, and MLVD were detected using immunohistochemically anticyclin E, HGF, CD34, and lymphatic vessel endothelial hyaluronan receptor 1 in 168 surgically resected ESCC cases and 30 normal esophageal mucosal samples. The expression levels of cyclin E, HGF, MVD, and MLVD were higher compared to controls. High cyclin E and HGF expression was found more frequently in the tumors larger than 5 cm (P < .001), with poorer differentiation (P = .034) and higher tumor node metastasis (TNM) staging (P = .009) compared to their counterparts. Both MVD and MLVD values were found to be higher in the tumors larger than 5 cm (P < .001), with poorer differentiation (P < .001) and higher TNM staging (P < .001) compared to their counterparts. Furthermore, the expression of MVD and MLVD in both the high cyclin E and high HGF expression groups was significantly higher compared to the low cyclin E and HGF expression groups (P < .001). This study demonstrated that high cyclin E and HGF expression is closely correlated with tumor size, tumor differentiation degree, and TNM stage in patients with ESCC. These findings proposed that cyclin E and HGF could serve as novel molecular markers for preoperational evaluation of ESCC.

Clinical characteristics and prognosis of ground-glass opacity nodules in young patients.

BACKGROUND: The detection rate of ground-glass opacity (GGO) in young patients has increased year by year with the increasingly widespread use of high-resolution computed tomography (HRCT) and the increased resolution of HRCT imaging. However, no scholars have reported the clinical characteristics and prognosis of GGO in young patients systematically. The purpose of this study is to investigate the clinical characteristics and prognosis presenting as GGO in young patients. METHODS: Clinical data of 127 young patients who were diagnosed as GGO and who underwent video-assisted thoracoscopic surgery (VATS) and had routine pathological examination were collected from January 2016 to January 2017. Nodules were classified according to benign and malignant: 26 benign nodules (Group A) and 115 malignant nodules (Group B). The pathological types, nodules size, surgical methods were analyzed, and the clinical characteristics and prognosis were evaluated. RESULTS: The results of pathological examination of 91 pure ground-glass opacities (pGGOs) revealed 16 adenocarcinoma in situs (AISs), 42 micro invasive adenocarcinomas (MIAs), 13 invasive adenocarcinomas (IAs), 8 atypical adenomatous hyperplasias (AAHs), 1 inflammatory granuloma, 2 pulmonary inflammatory pseudotumors (IPTs) and 9 other benign nodules. The results of pathological examination of 50 mixed ground-glass opacities (mGGOs) revealed 6 AISs, 29 MIAs, 9 IAs, 1 AAH, 2 inflammatory granulomas and 3 other benign nodules. All patients had no lymph nodes invasion. The rates of perioperative complications were 6.30%, compared to 7.63% for long-term complications. None of the patients with GGO experienced a recurrence and death [2-year recurrence-free survival (RFS), 100%; 2-year overall survival (OS), 100%]. CONCLUSIONS: The GGO in young patients that received VATS has a high proportion of malignant, its prognosis is satisfied.

TINCR facilitates non-small cell lung cancer progression through BRAF-activated MAPK pathway.

Long non-coding RNAs are critically involved in oncogenesis in various cancer types. Here we reported a novel lncRNA signature correlated with progression of non-small cell lung cancer (NSCLC). In particular, we identified elevated expression of terminal differentiation-induced noncoding RNA (TINCR) in human NSCLC samples, which were associated with enhanced migration, viability in NSCLC cells in vitro. Higher TINCR level was also correlated with poor survival. Furthermore, TINCR increased xenograft tumor growth in vivo mouse models. Mechanistic study demonstrated that TINCR can interact with BRAF to facilitate its kinase activity, thereby leading to activation of oncogenic mitogen-activated protein kinase (MAPK) pathway. These results suggested that TINCR upregulation may signal through the MAPK pathway to promote NSCLC tumorigenesis. Therefore, TINCR may serve as a potential prognostic marker and therapeutic target for NSCLC patients.

Antitumor activity of cobrotoxin in human lung adenocarcinoma A549 cells and following transplantation in nude mice.

The aim of the present study was to investigate cobra neurotoxin (cobrotoxin) activity in A549 cell lines transplanted into nude mice, and to explore its molecular mechanism. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used to detect the growth inhibition rate of cobrotoxin in human lung A549 adenocarcinoma cells and HFL1 lung fibroblasts. Cell colony formation assays were performed to determine the effect of cobrotoxin on A549 cell colony formation, and transmission electron microscopy was used to detect cobrotoxin autophagy. In addition, western blot analysis was performed to determine the effect of 3-methyl adenine (3-MA) activity on the inhibition of autophagy, SB203580 inhibition of the p38-mitogen-activated protein kinase (MAPK) pathway, and Beclin 1, LC3, p62, p38 and phosphorylated (p)-p38 protein expression. Nude mice were injected with human lung A549 cells, and intervention and control groups were compared with regard to tumor suppression. The MTT assay revealed that various concentrations of cobrotoxin inhibited growth of A549 cells, but not HFL1 cells. A549 cell colony formation decreased and autophagosome activity was significantly increased compared with the controls. Following 3-MA administration, SB203580 autophagosome activity decreased, and following cobrotoxin administration, Beclin 1, p-p38, and LC3-II protein expression significantly increased, whereas p62 expression significantly decreased. Following 3-MA inhibition of autophagy, Beclin 1, LC3-II and p62 expression increased. Furthermore, following SB203580 inhibition of the p38-MAPK pathway, Beclin 1, p-p38, LC3-II and p62 protein expression increased. Cobrotoxin exhibited inhibitory activity on the human lung cancer A549 cells transplanted into the nude mice, suppressing the tumor growth rate by 43.4% (cobrotoxin 40 µg/kg group). However, following the addition of 3-MA (10 mmol/kg) and SB203580 (5 mg/kg), the suppression of the tumor growth rate decreased significantly. Cobrotoxin inhibits the growth of human lung cancer A549 cells in vitro and A549 cells transplanted into nude mice. Furthermore, the induction of autophagy may be associated with the activation of the p38-MAPK pathway.

An albumin-based theranostic nano-agent for dual-modal imaging guided photothermal therapy to inhibit lymphatic metastasis of cancer post surgery.

A large variety of cancers are associated with a high incidence of lymph node metastasis, which leads to a high risk of cancer death. Herein, we demonstrate that multimodal imaging guided photothermal therapy can inhibit tumor metastasis after surgery by burning the sentinel lymph nodes (SLNs) with metastatic tumor cells. A near-infrared dye, IR825, is absorbed onto human serum albumin (HSA), which is covalently linked with diethylenetriamine pentaacetic acid (DTPA) molecules to chelate gadolinium. The formed HSA-Gd-IR825 nanocomplex exhibits strong fluorescence together with high near-infrared (NIR) absorbance, and in the mean time could serve as a T1 contrast agent in magnetic resonance (MR) imaging. In vivo bi-modal fluorescence and MR imaging uncovers that HSA-Gd-IR825 after being injected into the primary tumor would quickly migrate into tumor-associated SLNs through lymphatic circulation. Utilizing the strong NIR absorbance of HSA-Gd-IR825, SLNs with metastatic cancer cells can be effectively ablated under exposure to a NIR laser. Such treatment when combined with surgery to remove the primary tumor offers remarkable therapeutic outcomes in greatly inhibiting further metastatic spread of cancer cells and prolonging animal survival. Our work presents an albumin-based theranostic nano-probe with functions of multimodal imaging and photothermal therapy, together with a 'photothermal ablation assisted surgery' strategy, promising for future clinical cancer treatment.

[Analysis of single-operation-hole thoracoscopic lobectomy in 113 clinical cases].

BACKGROUND AND OBJECTIVE: Video-assisted thoracoscopic surgery (VATS) lobectomy is generally accepted for patients with lung cancer. The aim of this study is to explore the feasibility of the single-operation-hole thoracoscopic lobectomy in the treatment of non-small cell lung cancer. METHODS: To review and analyze the single-operation-hole thoracoscopic lobectomy performed in our hospital for 113 non-small cell lung cancer (NSCLC) cases from October 2010 to October 2013. The incision for observation was 1.5 cm the eighth intercostal at the rear of the midaxillary line and the incision for operation was 2.0 cm-4.0 cm at the fourth or fifth intercostal of the anterior axillary line. The operations were performed through the single-operation-hole. RESULTS: The operation processes were smooth for all the patients without any operative mortality occurrence. Only in 5 cases was the operation hole expanded because of the occurrence of massive hemorrhage during the operation; 3 patients with postoperative complications underwent thoracoscopic lobectomy again, including 2 cases of delayed hemorrhage and 1 case of chylothorax. The average surgical duration was (178.24±31.17) min, the average blood loss was (213.56±62.38) mL, and the number of lymph nodes dissected was from 5-22. All diagnose were confirmed by pathology after operation. The average length of stay was (8.17±2.93) d. All cases recovered well during the follow-up of (2-38) months, only 5 cases had recurrence or metastasis. CONCLUSIONS: The single-operation-hole thomcoscopic lobectomy for lung cancer is safe and feasible, further reducing the trauma, and can be used as a conventional treatment for early- or medium-term NSCLC.

PIK3CA hypomethylation plays a key role in activation of the PI3K/AKT pathway in esophageal cancer in Chinese patients.

AIM: To investigate the role of PIK3CA oncogene in tumorigenesis and development of esophageal cancer in Chinese patients at the levels of genetic mutation and epigenetics. METHODS: Seventy six esophageal tumor samples and corresponding adjacent normal tissues were collected, and the genomic DNA was extracted. Mutations in the 9th and 20th exons of PIK3CA gene were detected using conventional sequencing. PIK3CA methylation rates in two selected CpG islands (CpG island 1 and 2) were detected using sub-bisulfate modified sequencing. P110α and pAKT expression levels were detected with Western blotting. RESULTS: In PIK3CA gene of the tumor tissues, G1633C (E545Q) mutation was detected in the 9th exon with a rate of 3.95% (3/76), whereas mutation was not found in the 20th exon. Nor mutation did occur in PIK3CA gene of the adjacent normal tissues. The methylation rate of the CpG island 1 had no significant difference between the tumor and adjacent tissues (0.77%±0.009% vs 0.89%±0.008%), but the methylation rate of the CpG island 2 in the esophageal tumors was significantly lower than that in the adjacent tissues (6.00%±2.80% vs 10.45%±5.51%). Furthermore, the rate of methylation of the CpG island 2 in TNM stage III and IV esophageal cancer (3.84%±2.08%) was significantly lower than in stage I (8.52%±2.55%) and stage II (6.42%±2.36%). PIK3CA gene hypomethylation in esophageal cancer was significantly correlated with high expression of p110α. CONCLUSION: PIK3CA gene hypomethylation plays a key role in the tumorigenesis and development of esophageal cancer in Chinese patients, while the mutations of PIK3CA gene have little effect on the development of esophageal cancer.

[Clinical analysis of thoracoscopic lobectomy in the treatment of peripheral lung cancer with single utility port].

BACKGROUND AND OBJECTIVE: Video-assisted thoracoscopic surgery (VATS) lobectomy is now generally accepted for patients with lung cancer. The aim of this study is to review the technology of thoracoscopic lobectomy with single utility port in the treatment of peripheral lung cancer. METHODS: We retrospectively analyzed the clinical data of 87 patients with peripheral lung cancer who underwent single utility port complete VATS lobectomy from February 2011 to January 2013 in the First Affiliated Hospital of Soochow University (single utility port group), and compared them with 75 patients with peripheral lung cancer who underwent conventional, 3-port VATS lobectomy in the same period (3-port group). The clinical outcomes including operation time, time to first activity out of bed, postoperative hospital stay, intraoperative blood loss, postoperative drainage volume, chest drainage duration, lymph node dissection number, postoperative complications and degree of chest pain were compared between the two groups. RESULTS: No perioperative death was observed in both groups. There was no statistical difference in operation time (151.03±25.97 min vs 156.27±26.49 min), lymph node dissection number (13.06±1.36 vs 12.61±1.56), intraoperative blood loss (188.62±47.03 mL vs 179.60±28.96 mL) and incidence of serious postoperative complications (18/87 vs 21/75) between the two groups. There were statistical differences in time to first activity out of bed (11.17±8.69 h vs 13.76±7.43 h), postoperative hospital stay (7.18±1.95 d vs 7.92±2.03 d), chest drainage duration (3.85±1.21 d vs 4.43±1.43 d) and total postoperative drainage volume (671.49±178.31 mL vs 736.93±170.39 mL) between the two groups (P<0.05). The change of vision analogue score (VAS) score between the two groups after operation was also statistically significant (P<0.01). CONCLUSIONS: The completely thoracoscopic lobectomy with single utility port is a safe and feasible surgical procedure compared with conventional 3-port VATS lobectomy for selected patients.

[Role of autophagy on cobrotoxin induced cell death of A549].

BACKGROUND: It has been proven that cobrotoxin has anti-tumor effect while its role in lung cancer is rarely studied. The aim of this study is to assay the anti-tumor effect of cobrotoxin in cell line A549, and also to explore its possible mechanism related to autophagy and P38-MARK pathway. METHODS: Using MTT method to observe the inhibition effect of cobrotoxin on the growth of adenocarcinoma cell A549 and human lung fibroblast cell HFL1, as well as on that of A549 pretreated with 3-MA and SB203580, which are the inhibitor of autophagy and P38-MARK pathway respectively. Cell colony tablet cloning experiment was executed to detect the effect of cobrotoxin on colony formation of A549. Determining the protein levels of beclin-1, LC3, p38 and pP38 in A549 by Western blot after cells were exposed to cobrotoxin, or to cobrotoxin combined with either 3-MA or SB203580. RESULTS: All of different concentrations of cobrotoxin inhibited the growth of A549, but had no obvious effect on that of HFL1. After treating with 3-MA or SB203580, the suppress effect of cobrotoxin on A549 reduced. What's more, different concentrations of cobrotoxin all significantly suppressed the colony formation of A549. The expression of beclin-1 and pP38 in A549 increased obviously after exposure to cobrotoxin, and also the ratio of LC3 II to LC3 I amplified with a dose-dependant manner, but P62 decreased. The protein level of beclin-1 and the ratio of LC3 II to LC3 I in cells pretreated with 3-MA were reduced, while that of p62 was increased. Also, in cells that treated with SB203580 before exposed to cobrotoxin, the expression of beclin-1, pP38 and the ratio of of LC3 II to LC3 I were reduced, but the expression of P62 increased. CONCLUSIONS: Cobrotoxin can suppress the growth of A549 in vitro. And the activating of P38-MARK pathway, then upregulating autophagy, was involved in cobrotoxin induced anti-tumor process.

Growth inhibitory effects and molecular mechanisms of crotoxin treatment in esophageal Eca-109 cells and transplanted tumors in nude mice.

AIM: To investigate the antitumor actions of the Crotalus durissus neurotoxin (crotoxin) on human esophageal carcinoma (Eca-109) cells in vitro and transplanted esophageal Eca-109 tumors in nude mice. METHODS: The growth-inhibitory effect was analyzed in Eca-109 cells using MTT assay. Cell morphology changes in nuclei were observed using Hoechst 33342 staining, while apoptosis and cell cycle distribution were examined by flow cytometry. RT-PCR was used to measure the Bcl-2, p15, and caspase-3 p17 gene expression levels. A tumor transplantation model was established by inoculation of Eca-109 cells were into female Balb/c nude mice. Crotoxin (25, 50, and 100 mg/kg) was subcutaneously injected into the transplanted tumors every 2 d for a total of 10 injections. Tumor size and weight were measured. Bcl-2, p15, and caspase-3 p17 protein expression in transplanted tumors was analyzed using Western blotting. RESULTS: Crotoxin (25, 50, and 100 µg/mL) inhibited the growth of Eca-109 cells in a dose-dependent manner with inhibition rates of 22.9%, 35.8%, and 57.2%, respectively. Hoechst 33342 staining revealed apoptotic cells with pyknotic nuclear chromatin after crotoxin treatment. In Eca-109 cells, crotoxin induced apoptosis and G1 block, significantly upregulated the expression of p15 and caspase-3 p17 genes and downregulated the expression of Bcl-2 gene. Furthermore, crotoxin inhibited the growth of Eca-109 tumors in nude mice in a dose-dependent manner. Western blotting showed that crotoxin increased p15 and caspase-3 p17 protein levels and reduced Bcl-2 protein level in tumor specimens. CONCLUSION: Crotoxin inhibits the growth of Eca-109 cells in vitro via apoptosis induction and G1 block. Local administration of crotoxin inhibits the growth of subcutaneously transplanted Eca-109 cells in nude mice, possibly via increasing p15 and caspase-3 p17 protein expression and reducing Bcl-2 protein expression.

Fully thoracoscopic versus conventional open resection for esophageal carcinoma: A perioperative comparison.

BACKGROUND: To compare the efficacy of patients undergoing esophagectomy for cancer in video-assisted thoracoscopic surgery (VATS) versus traditional open surgery (TOS) in the perioperative period, along with the advantages and disadvantages of each. METHODS: A retrospective analysis of 108 patients, who underwent esophagectomy between September 2011 and February 2012 in our department, was performed. Patients were divided into two groups based on operative technique (VATS vs. TOS), with 50 patients in the VATS group and 58 patients in the TOS group. Operative duration, intraoperative blood loss, intraoperative blood transfusion, number of lymph nodes harvested, postoperative pain score, period of time requiring chest tube drainage, complications, hospital stay, and hospital costs, were all statistically analyzed between the two groups. RESULTS: There was no statistical difference between the two groups with regard to operative duration or number of lymph nodes harvested. The VATS group had significantly less intraoperative blood loss, intraoperative blood transfusion, postoperative pain, earlier ambulation, shorter postoperative hospital stay, and a shorter period of time requiring chest tube drainage. The amount of drainage was significantly lower in the TOS group (P < 0.05). Pulmonary complication (pneumonia and pleural effusion) was less prevalent among the VATS group. CONCLUSION: Compared with TOS, VATS-assisted esophagectomy is less traumatic with lower intraoperative blood loss, faster recovery, and a better overall outcome.

Crotoxin enhances the antitumor activity of gefinitib (Iressa) in SK-MES-1 human lung squamous carcinoma cells.

Crotoxin (CrTX), a neurotoxin, is isolated from the venom of South American rattlesnakes and has potent antitumor activity. Here, we investigated the antitumor effect of CrTX on the SK-MES-1 human lung squamous cell carcinoma cell line that has acquired resistance to epidermal growth factor receptor tyrosine kinase inhibitors. CrTX caused G1 arrest and p-JNK protein upregulation that resulted in apoptosis of SK-MES-1 cells. SP600125, a specific inhibitor of p-JNK, could rescue SK-MES-1 cells from CrTX-induced apoptosis. CrTX and gefinitib (Iressa) both inhibited the viability and proliferation of SK-MES-1 cells in a dose- and time-dependent manner. The combination of CrTX and Iressa significantly enhanced the antitumor activity of Iressa. In vivo studies revealed that CrTX caused increased damage to blood vessels and reduced tumor size when combined with Iressa. The present study showed that the JNK signal transduction pathway mediated the anti-apoptotic effect of CrTX, and furthermore, CrTX could enhance the antitumor effect of tyrosine kinase inhibitors in cells with acquired resistance.

Anti-tumor activity of CrTX in human lung adenocarcinoma cell line A549.

AIM: To assess the cytotoxic effect of crotoxin (CrTX), a potent neurotoxin extracted from the venom of the pit viper Crotalus durissus terrificus, in human lung adenocarcinoma A549 cells and investigated the underlying mechanisms. METHODS: A549 cells were treated with gradient concentrations of CrTX, and the cell cycle and apoptosis were analyzed using a flow cytometric assay. The changes of cellular effectors p53, caspase-3 and cleaved caspase-3, total P38MAPK and pP38MAPK were investigated using Western blot assays. A549 xenograft model was used to examine the inhibition of CrTX on tumor growth in vivo. RESULTS: Treatment of A549 cells with CrTX (25-200 µg/mL) for 48 h significantly inhibited the cell growth in a dose-dependent manner (IC(50)=78 µg/mL). Treatment with CrTX (25 µg/mL) for 24 h caused G1 arrest and induced cell apoptosis. CrTX (25 µg/mL) significantly increased the expression of wt p53, cleaved caspase-3 and phospho-P38MAPK. Pretreatment with the specific P38MAPK inhibitor SB203580 (5 µmol/L) significantly reduced CrTX-induced apoptosis and cleaved caspase-3 level, but G(1) arrest remained unchanged and highly expressed p53 sustained. Intraperitoneal injection of CrTX (10 µg/kg, twice a week for 4 weeks) significantly inhibited A549 tumor xenograft growth, and decreased MVD and VEGF levels. CONCLUSION: CrTX produced significant anti-tumor effects by inducing cell apoptosis probably due to activation of P38MAPK and caspase-3, and by cell cycle arrest mediated by increased wt p53 expression. In addition, CrTX displayed anti-angiogenic effects in vivo.

Triptolide inhibits NF-kappaB activation and reduces injury of donor lung induced by ischemia/reperfusion.

AIM: To investigate the protective effect of triptolide (TRI) on ischemia/reperfusion-induced injury of transplanted rabbit lungs and to investigate the mechanisms underlying the actions of TRI. METHODS: We established the rabbit lung transplantation model and studied lung injury induced by ischemia/reperfusion and the inhibitory effect of TRI on NF-kappaB. The severity of lung injury was determined by a gradual decline in PvO2, the degree of lung edema, the increase in the myeloperoxidase (MPO) activity, and the ultrastructural changes of transplanted lungs. The activation of NF-kappaB was measured by immunohistochemistry. The increase in intercellular adhesion molecule-1 (ICAM-1), which is the target gene of NF-kappaB, was evaluated by ELISA. RESULTS: After reperfusion, there was a gradual decline in the PvO2 level in the control group (group I). The level of PvO2 in the group treated with lipopolysaccharide (group II) was significantly decreased, whereas that of the group treated with TRI (group III) was markedly improved (P<0.01). In group III, the activity of MPO was downregulated, and the pulmonary edema did not become severe and the ultrastructure of the donor lung remained normal. The activity of NF-kappaB and the expression of ICAM-1 was significantly increased in the donor lungs. TRI blocked NF-kappaB activation and ICAM-1 expression. CONCLUSION: The effects of TRI on reducing injury to donor lungs induced by ischemia/reperfusion may possibly be mediated by inhibiting the activity of NF-kappaB and the expression of the NF-kappaB target gene ICAM-1. Thus, TRI could be used in lung transplantations for improving the function of donor lungs.

Inhibitory effects of prostaglandin A1 on apoptosis of rat cardiac microvascular endothelial cells was mediated by NF-kappaB.

AIM: To study the effects of prostaglandin A1 (PGA1) on rat cardiac microvascular endothelial cells. METH-ODS: Isolated rat cardiac microvascular endothelial cells were cultured in hypoxia and reoxygen conditions, respectively. Endothelial cell apoptosis was detected by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling staining. The activity of NF-kappaB was detected by electrophoretic mobility shift assay. Bcl-2 and Bax protein expression were examined by Western blot and bcl-2 mRNA expression was examined by Northern blot. RESULTS: PGA1 reduced endothelial cell apoptosis markedly, inhibited activity of NF-kappaB, and increased expression of Bcl-2 protein and bcl-2 mRNA. However, PGA1 did not alter Bax protein expression resulting in an increase in the ratio of Bcl-2 to Bax. CONCLUSION: PGA1 can inhibit rat cardiac microvascular endothelial cell apoptosis by inhibiting activity of NF-kappaB.