Human umbilical cord mesenchymal stem cell-derived extracellular vesicles carrying miR-655-3p inhibit the development of esophageal cancer by regulating the expression of HIF-1α via a LMO4/HDAC2-dependent mechanism.

OBJECTIVE: This study clarified the function of human umbilical cord mesenchymal stem cell (hUCMSC)-derived extracellular vesicle (EV)-enclosed miR-655-3p in esophageal squamous cell carcinoma (ESCC). METHODS: A Chi-square test and the Kaplan-Meier estimator were used to analyze the prognosis of ESCC in relation to the expression of miR-655-3p. ESCC cells were incubated with PBS or hUCMSC-derived EVs (hUCMSC-EVs) in the conditions of gene modification, after which the malignant behaviors of ESCC cells were assessed and the molecular interactions were determined. The effect of hUCMSC-derived EV-miR-655-3p was also investigated in a nude mouse model of ESCC. RESULTS: Low expression of miR-655-3p indicated poor prognosis of ESCC. hUCMSC-EVs suppressed the malignant behaviors of ESCC cells and the growth and liver metastasis of transplanted tumors. Inhibition of miR-655-3p in hUCMSCs impaired the therapeutic effect of hUCMSC-EVs. LMO4, targeted by miR-655-3p, activated the transcription of HIF-1α by sequestering HDAC2 from HIF-1α promoter. Knockdown of LMO4 suppressed ESCC cell activities, while overexpression of HIF-1α counteracted the tumor suppressive effect of LMO4 knockdown. CONCLUSION: miR-655-3p enclosed in hUCMSC-derived EVs inhibits ESCC progression partially by inactivating HIF-1α via the LMO4/HDAC2 axis.

Comprehensive characterization of genomic and radiologic features reveals distinct driver patterns of RTK/RAS pathway in ground-glass opacity pulmonary nodules.

Ground-glass opacity (GGO)-associated pulmonary nodules have been known as a radiologic feature of early-stage lung cancers and exhibit an indolent biological behavior. However, the correlation between driver genes and radiologic features as well as the immune microenvironment remains poorly understood. We performed a custom 1021-gene panel sequencing of 334 resected pulmonary nodules presenting as GGO from 262 Chinese patients. A total of 130 multiple pulmonary nodules were sampled from 58 patients. Clinical-pathologic and radiologic parameters of these pulmonary nodules were collected. Immunohistochemistry (IHC) and multiplex immunofluorescent staining (mIF) were applied to analyze proliferation and immune cell markers of GGO-associated pulmonary nodules. Compared with pure GGO nodules, mixed GGO nodules were enriched for invasive adenocarcinoma (IAC) (182/216 vs 73/118, P < .001). Eighty-eight percent (294/334) of GGO-associated nodules carried at least one mutation in EGFR/ERBB2/BRAF/KRAS/MAP2K1 of the RTK/RAS signaling pathway, and the alterations in these driver genes were mutually exclusive. The analysis of multifocal pulmonary nodules from the same patient revealed evidence of functional convergence on RTK/RAS pathways. Nodules with ERBB2/BRAF/MAP2K1 mutations tended to be more indolent than those with EGFR and KRAS mutations. IHC and mIF staining showed that KRAS-mutant GGO nodules displayed higher infiltration of CD4+ T cell and CD8+ T cell as well as stronger proliferation and immune inhibitory signals. Our study demonstrates a driver landscape of radiologically detectable GGO-associated pulmonary nodules in Chinese patients and supports that different driver patterns in RTK/RAS pathway are corresponding to different radiologic features.

Early stage of antibody-mediated rejection after lung transplantation： A case report and literature review. / 肺移植术后早期抗体介导排斥反应1ä¾‹åŠæ–‡çŒ®å¤ä¹ .

Antibody-mediated rejection (AMR) is a rare and serious complication after lung transplantation, with no characteristic of pathological manifestation, no systematic standard treatment, and the poor efficacy and prognosis. We reported a case of early AMR after lung transplantation and the relevant literature has been reviewed. A male patient presented with symptoms of cold 99 days after transplantation and resolved after symptomatic treatment. He admitted to the hospital 14 days later because of a sudden dyspnea and fever. Anti-bacteria, anti-fungi, anti-virus, and anti-pneumocystis carinii treatment were ineffective, and a dose of 1 000 mg methylprednisolone did not work too. The patient's condition deteriorated rapidly and tracheal intubation was done to maintain breathing. Serum panel reactive antibody and donor specific antibody showed postive in humen leukocyte antigen (HLA) II antibody. Pathological examination after transbronchial transplantation lung biopsy showed acute rejection. Clinical AMR was diagnosed combined the donor-specific antibody with the pathological result. The patient was functionally recovered after combined treatment with thymoglobuline, rituximab, plasmapheresis, and immunoglobulin. No chronic lung allograft dysfunction was found after 3 years follow up. We should alert the occurrence of AMR in lung transplantation recipient who admitted to hospital with a sudden dyspnea and fever while showed no effect after common anti-infection and anti-rejection treatment. Transbronchial transplantation lung biopsy and the presence of serum donor-specific antibody are helpful to the diagnosis. The treatment should be preemptive and a comprehensive approach should be adopted.

[Tumor infiltrating T lymphocyte components in malignant pleural effusion of lung adenocarcinoma and their killing activities to autologous tumor cells].

OBJECTIVE: To analyze the components of tumor infiltrating T lymphocyte (TIL) cells in malignant pleural effusion of lung adenocarcinoma, and evaluate their killing activities to autologous tumor cells. 
 Methods: Malignant pleural effusions were collected from 17 patients with lung adenocarcinoma. Mononuclear cells were isolated by Ficoll density gradient centrifugation and flow cytometer was used to analyze TIL cell components. TIL and tumor cells were separated through adherent culture. The tumor cells were identified via intramuscular injection of adherent cells into nude mice and the killing effect of cultured lymphocytes on autologous tumor cells was studied.
 Results: Of the TIL in malignant pleural effusions, T cells accounted for 60.6%-79.3%, while T helper cells were significantly higher than T killer cells (36.63%±1.90% vs 24.64%±2.32%, P<0.001). There were also natural killer (NK) cells and NK T cells in the effusions. Tumor cells were successfully isolated and cultured. The killing activity of cultured TIL to autologous tumor cells was 39.14%±12.04%, and the killing activity of TIL with high proliferation rate to autologous tumor cells was higher than that of low proliferation group (50.51%±3.80% vs 29.04%±5.77%, P<0.001).
 Conclusion: T lymphocytes are the major components of TIL in malignant pleural effusions derived from lung adenocarcinoma, and T helper cells are more than T killer cells. The killing activity of TIL with strong proliferation ability to autologous tumor cells is higher than that of TIL with weak proliferation ability. Therefore, cells from malignant pleural effusions could be used for cellular immunotherapy against tumor.

LncRNA MALAT1 promotes epithelial-to-mesenchymal transition of esophageal cancer through Ezh2-Notch1 signaling pathway.

To investigate effect of long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) on epithelial-to-mesenchymal transition (EMT) of esophageal cancer (EC) and role of enhancer of zeste homolog 2 (Ezh2)-Notch1 signaling pathway in the process. The expression of MALAT1 was determined in four EC cell lines by real-time PCR. TE-1 and EC109 cells were transfected with sh-MALAT1 to inhibit expression of MALAT1 or transfected with pcDNA3.1-Ezh2 to overexpress Ezh2. Invasion and migration assays were conducted to analyze cell metastasis, and expressions of Ezh2-Notch1 signaling-related proteins as well as EMT related proteins were determined using both real-time PCR and western blot. MALAT1 was significantly up-regulated in all EC cell lines compared with the normal cells. Silencing MALAT1 using shRNA could significantly inhibit cell viability (reduced almost 30% of cell viability compared with the control), invasion (reduced almost 30% of cell migration compared with the control), and migration (reduced almost 50% of cell migration compared with the control) of both TE-1 and EC109 cells (P<0.05). Meanwhile, expression of Ezh2, Notch1, Hes1, MMP-9, and Vimentin was significantly decreased and expression of E-cadherin was significantly increased when cells were transfected with sh-MALAT1 compared with the nontransfected cells (P<0.05). However, when cells were cotransfected with both sh-MALAT1 and pcDNA3.1-Ezh2, the protein expression changes induced by sh-MALAT1 were recovered. MALAT1 could affect EMT and metastasis of EC cells through Ezh2-Notch1 signaling pathway. This study can give deeper understandings of the role of MALAT1 in EC and may provide some new directions for treatment of patients with EC.

[Serum levels of miRNA-183 in patients with esophageal squamous cell carcinoma and its diagnostic value].

OBJECTIVE: To explore the changes of serum microRNA-183 levels in patients with esophageal squamous cell carcinoma (ESCC) and its clinical significance.
 Methods: Fifty-one patients with ESCC and 55 healthy subjects from Department of Cardiothoracic Surgery, Second Xiangya Hospital, Central South Unicersity were selected for this study. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to determine the level of miRNA-183 in serum samples. Chi-square test and correlation analysis were used to investigate the relationship between serum miRNA-183 level and clinical and pathological parameters of ESCC. Diagnostic efficiency of miRNA-183 and combined carcinoembryonic antigen (CEA) examination for ESCC was analyzed by receiver operating characteristic (ROC) curve.
 Results: 1) The levels of miR-183 in the patients with ESCC (4.47±1.54) were elevated compared with that in the healthy subjects (2.03±0.96), with significant difference (t=9.700, P<0.01). 2) The levels of serum miR-183 in ESCC patients were significantly different among patients with different TNM stages (χ2=4.049, P<0.01), which was not affected by gender, age, smoking, drinking, tumor location, tumor diameter, lymph node metastasis, depth of invasion and differentiation (all P>0.05). The levels of miR-183 were not associated with the serum CEA levels (P>0.05). 3) When the ROC curve analysis was used to diagnose ESCC with the optimal cutoff value of 4.502 for miR-183, the sensitivity, the specificity, the area under the curve (AUC) and 95% confidence interval was 78.9%, 76.2%, 0.762 and 0.830-0.922, respectively. When combined detection of serum miR-183 and CEA was used to diagnose ESCC, the sensitivity, specificity, AUC and 95% confidence interval was 82.3%, 92.6%, 0.877 and 0.814-0.935, respectively.
 Conclusion: Serum miRNA-183 levels in ESCC patients may be increased, which can improve the diagnostic efficiency of ESCC when combined with CEA. Serum miRNA-183 levels is related with tumor TNM stage, which contributes to the judgment of tumor progression and efficacy prediction.

Regulation of Mcl-1 by constitutive activation of NF-κB contributes to cell viability in human esophageal squamous cell carcinoma cells.

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is one of the most lethal malignancies with a 5-year survival rate less than 15%. Understanding of the molecular mechanisms involved in the pathogenesis of ESCC becomes critical to develop more effective treatments. METHODS: Mcl-1 expression was measured by reverse transcription (RT)-PCR and Western blotting. Human Mcl-1 promoter activity was evaluated by reporter gene assay. The interactions between DNA and transcription factors were confirmed by electrophoretic mobility shift assay (EMSA) in vitro and by chromatin immunoprecipitation (ChIP) assay in cells. RESULTS: Four human ESCC cell lines, TE-1, Eca109, KYSE150 and KYSE510, are revealed increased levels of Mcl-1 mRNA and protein compare with HaCaT, an immortal non-tumorigenic cell line. Results of reporter gene assays demonstrate that human Mcl-1 promoter activity is decreased by mutation of kappaB binding site, specific NF-kappaB inhibitor Bay11-7082 or dominant inhibitory molecule DNMIkappaBalpha in TE-1 and KYSE150 cell lines. Mcl-1 protein level is also attenuated by Bay11-7082 treatment or co-transfection of DNMIkappaBalpha in TE-1 and KYSE150 cells. EMSA results indicate that NF-kappaB subunits p50 and p65 bind to human Mcl-1-kappaB probe in vitro. ChIP assay further confirm p50 and p65 directly bind to human Mcl-1 promoter in intact cells, by which regulates Mcl-1 expression and contributes to the viability of TE-1 cells. CONCLUSIONS: Our data provided evidence that one of the mechanisms of Mcl-1 expression in human ESCC is regulated by the activation of NF-kappaB signaling. The newly identified mechanism might provide a scientific basis for developing effective approaches to treatment human ESCC.

Pulmonary artery pseudoaneurysm caused by a rare vascular tumor: epithelioid hemangioendothelioma.

We present a case of a 58-year-old female with a rare vascular tumor of intermediate malignancy. The initial manifestation was a pseudoaneurysm caused by the rupture of the right pulmonary artery after tumor invasion. The diagnosis of epithelioid hemangioendothelioma was confirmed by the morphologic and immunocytochemical features after surgery. The patient recovered smoothly and there has been no evidence of local recurrence or metastasis during the 2 years of follow-up.

CHAC2 promotes lung adenocarcinoma by regulating ROS-mediated MAPK pathway activation.

An imbalance in ROS (reactive oxidative species) and the antioxidant barrier regulates the process of tumorigenesis. GSH has a key effect in preventing cells from oxidative damage by scavenging ROS. The role of CHAC2, an enzyme regulating GSH, in lung adenocarcinoma remains unknown. Here, RNA sequencing data analysis and immunohistochemistry (IHC) assays of lung adenocarcinoma and normal lung tissues were used to verify the expression of CHAC2. The effect of CHAC2 on the proliferation abilities of lung adenocarcinoma cells was examined using a series of overexpression or knockout assays. RNA sequencing and IHC results showed that the expression level of CHAC2 in lung adenocarcinoma was higher than that in normal lung tissues. CCK-8, colony formation and subcutaneous xenograft experiments in BALB/c nude mice showed that in vitro and in vivo CHAC2 promoted the growth capacity of lung adenocarcinoma cells. Subsequent immunoblot, immunohistochemistry and flow cytometry experiments showed that CHAC2 increased ROS by reducing GSH in lung adenocarcinoma and that the elevated ROS activated the MAPK pathway. Our investigation identified a new role for CHAC2 and elucidated the mechanism by which CHAC2 promotes lung adenocarcinoma progression.

[Evaluation of resection of local advanced upper lung cancer through median sternotomy].

OBJECTIVE: To summarize the resection of local advanced upper lung cancer and radical bilateral mediastinal lymph node dissection through a median sternotomy. METHODS: A total of 31 patients with local advanced upper lung cancer underwent lobectomy and radical complete dissection of bilateral superior mediastinal lymph node through a median sternotomy (the sternotomy group). The sternotomy group consisted of 8 females and 23 males, from 35 to 75 years old (average 57 years). Five patients underwent superior vena caval replacement or partial excision, 21 underwent upper sleeve lobectomy, and 6 patients combined with right pulmonary artery sleeve angioplasty or partial resection and reconstruction. Compared with the 30 patients who were operated through posterolateral incision, the surgery time, complications, and prognosis during the same period (the posterolateral incision group) were recorded. RESULTS: There was no perioperative death. The average operation time in the sternotomy group was (170±30)min, while that in the posterolateral incision group was (140±30) min(P>0.05). Postoperative complications comprised atelectasis, cardiac arrhythmia, and pneumonia. In the sternotomy group it was 6.5%(2/31), 16.1%(5/31), and 6.5% (2/31),and that in the posterolateral incision group 3.3%(1/30), 20%(6/30), 10.0%(3/30),respectively. Postoperative pathological findings demonstrated the rate for pN3 disease in the sternotomy group was 29%(9/31), 2 patients died of brain and liver metastasis respectively 10 or 11 months after the operation. The 3 year survival rate of 9 patients with pN3 diagnosed as cN2 preoperatively was 33.3%(3/9). The total survival rate of 1,3 years in the sternotomy group was 90.3%(28/31) and 41.9%(13/31), in the posterolateral incision group 86.6%(26/30) and 40.0%(12/30),respectively(P>0.05). CONCLUSION: Median sternotomy helps to resect local advanced upper lung cancer completely and to dissect bilateral mediastinal lymph node, and it can also provide more complete postoperative lymph node staging with no significant increase in complications.

[Application of laminated anastomosis with absorbable suture in cervical esophagogastrostomy].

OBJECTIVE: To observe the clinical results of laminated anastomosis using absorbable suture in cervical esophagogastrostomy, and to reduce the incidence of cervical esophagogastric anastomotic stricture. METHODS: A retrospective analysis was carried out on 210 patients who underwent cervical esophagogastrostomy after subtotal esophagectomy from January 2008 to June 2010. Among them, 96 cases were treated with traditional full layer interrupted varus suture (varus group) and the remaining 114 cases were treated with seromuscular layer and mucosal layer laminated anastomosis with absorbable suture (laminated group). Esophageal angiography was performed in 1 week, 1 month, and 3 months after the operation. The diameter of anastomatic stoma was measured on the anteroposterior and lateral angiography image respectively. The area of anastomatic stoma was calculated. The degree of stenosis was assessed according to the patients' dysphagia symptom. RESULTS: There was no operative deaths, no serious pulmonary complications and chylothorax, no sever esophageal reflux in all patients. The ratio of cervical esophagogastric anastomotic leakage was 2.1% (2/96) in the varus group. No anastomotic leakage in the laminated group. Compared with the varus group, the area of the anastomatic stoma in the laminated group was significantly increased in all measured time points (P<0.01). The incidence of obstruction in the laminated group was decreased significantly (P<0.01) in 1 month or in 3 months after operation compared with the varus group. CONCLUSION: Application of the laminated anastomosis with absorbable suture in cervical esophagogastrostomy can significantly reduce the incidence of anastomotic stenosis.

Identification of the Unique Clinical and Genetic Features of Chinese Lung Cancer Patients With EGFR Germline Mutations in a Large-Scale Retrospective Study.

BACKGROUND: Epidemiological surveys have suggested that lung cancer has inherited susceptibility and shows familial aggregation. However, the distribution and prevalence of epidermal growth factor receptor (EGFR) germline variants and their roles in lung cancer genetic predisposition in Chinese population remain to be elucidated. METHODS: In this study, EGFR germline and somatic variants were retrospectively reviewed from the next-generation sequencing results of 31,906 patients with lung cancer. Clinical information was also collected for patients with confirmed EGFR germline mutations. RESULTS: A total of 22 germline EGFR variants were identified in 64 patients with lung cancer, accounting for 0.2% of the total cases studied. Five patients were diagnosed as multiple primary carcinomas. Family history was documented in 31.3% (20/64) of patients, 55% of which were diagnosed as lung cancer. G863D was the most frequent EGFR germline mutation, followed by P848L, D1014N, and K757R. Somatic EGFR-sensitive mutations were identified in 51.6% of patients with germline EGFR mutations. The proportion of L858R mutation, exon 19 deletion, and rare sensitive mutation was 50%, 17.6%, and 32.4%, respectively. D1014N and T790M mutations were common in young patients. The family members of patients with P848L, R776H, V769M, and V774M mutations were more commonly diagnosed with cancers. A total of 19 patients were confirmed to have received EGFR tyrosine kinase inhibitors (TKIs), but the response to EGFR-TKIs differed among patients with different EGFR mutations. CONCLUSION: Chinese patients with lung cancer harbored unique and dispersive EGFR germline mutations and showed unique clinical and genetic characteristics, with varied response patterns to EGFR-TKI treatment.

A novel HER2-targeting antibody 5G9 identified by large-scale trastuzumab-based screening exhibits potent synergistic antitumor activity.

BACKGROUND: Pertuzumab is currently used in combination with trastuzumab as the first-line treatment for HER2-positive metastatic breast cancer. However, pertuzumab was originally developed independently from trastuzumab and was later incidentally found to have synergistic efficacy when combined with trastuzumab, it remains to be seen whether a more potent synergistic efficacy partner exists for trastuzumab. METHODS: A trastuzumab-based functional assay was used to screen anti-HER2 antibodies harboring trastuzumab-synergistic antitumor activity. The lead candidate 5G9, in combination with trastuzumab, was further characterized for its bioactivities in cell proliferation, cell apoptosis, antigen-antibody endocytosis and HER2-mediated cell signaling pathway blocking. Finally, animal models were used to evaluate the in vivo synergistic antitumor efficacy of 5G9 in combination with trastuzumab. FINDINGS: Compared to pertuzumab, 5G9 demonstrated more potent synergistic cell growth inhibitory activity when combined with trastuzumab (85% vs. 55%, P<0.001). In addition, 5G9 exhibited a higher internalization rate than pertuzumab (20% vs. 9%, P<0.05), and was able to further synergize with trastuzumab to promote antigen-antibody endocytosis. The internalization rate of the combination of 5G9 and trastuzumab was higher than that of pertuzumab and trastuzumab (35% vs. 14%, P<0.001). In vivo animal studies demonstrated that 5G9 in combination with trastuzumab showed more potent synergistic antitumor efficacy than the combination of pertuzumab and trastuzumab. INTERPRETATION: 5G9, together with trastuzumab, may provide a potential opportunity for more efficacious treatment of HER2-positive cancers. FUNDING: National Natural Science Foundation of China; State Key Laboratory of Analytical Chemistry for Life Science.

Ultrasensitive DNA hypermethylation detection using plasma for early detection of NSCLC: a study in Chinese patients with very small nodules.

PURPOSE: We had previously developed highly sensitive DNA methylation detection to diagnose lung cancer in patients with pulmonary nodules. To validate this approach and determine clinical utility in Chinese patients with indeterminate pulmonary nodules, we assessed the diagnostic accuracy for early stage lung cancer in plasma samples. EXPERIMENTAL DESIGN: Patients with CT-detected small lung nodules (diameter ≤ 3.0 cm) were included. Cases (n = 163) had staged IA or IB non-small cell lung cancer (NSCLC), while controls (n = 83) had non-cancerous lesions. Promoter methylation of eight lung cancer-specific genes (CDO1, TAC1, SOX17, HOXA7, HOXA9, GATA4, GATA5, and PAX5) was detected using nanoparticle-based DNA extraction (MOB) followed by qMSP. RESULTS: Methylation detection for CDO1, TAC1, SOX17, and HOXA7 in plasma was significantly higher in cases compared with the benign group (p < 0.001). The sensitivity and specificity for lung cancer diagnosis using individual gene was 41-69% and 49-82%. A three-gene combination of the best individual genes has sensitivity and specificity of 90% and 71%, with area under the receiver operating curve (AUC) of 0.88, (95% CI 0.84-0.93). Furthermore, three-gene combinations detected even the smallest lung nodules, with the combination of CDO1, SOX17, and HOXA7 having the overall best performance, while the combination of CDO1, TAC1, and SOX17 was best in tumor sizes less than 1.0 cm. CONCLUSIONS: Using modified MOB-qMSP, high sensitivity and specificity, for the detection of circulating tumor DNA was obtained for early stage NSCLC. This strategy has great potential to identify patients at high risk and improve the diagnosis of lung cancer at an earlier stage.

Biochemical and cellular characteristics of the 3' -> 5' exonuclease TREX2.

TREX2 is an autonomous nonprocessive 3' --> 5' exonuclease, suggesting that it maintains genome integrity. To investigate TREX2's biochemical and cellular properties, we show that endogenous TREX2 is expressed widely in mouse tissues and human cell lines. Unexpectedly, endogenous human TREX2 is predominantly expressed as a 30-kDa protein (not 26 kDa, as previously believed), which is likely encoded by longer isoforms (TREX2(L1) and/or TREX2(L2)) that possess similar capacity for self-association, DNA binding and catalytic activity. Site-directed mutagenesis analysis shows that the three functional activities of TREX2 are distinct, yet integrated. Mutation of amino acids putatively important for homodimerization significantly impairs both DNA binding and exonuclease activity, while mutation of amino acids (except R163) in the DNA binding and exonuclease domains affects their corresponding activities. Interestingly, however, DNA-binding domain mutations do not impact catalytic activity, while exonuclease domain mutations diminish DNA binding. To understand TREX2 cellular properties, we find endogenous TREX2 is down regulated during G2/M and nuclear TREX2 displays a punctate staining pattern. Furthermore, TREX2 knockdown reduces cell proliferation. Taken together, our results suggest that TREX2 plays an important function during DNA metabolism and cellular proliferation.

Cisplatin depletes TREX2 and causes Robertsonian translocations as seen in TREX2 knockout cells.

Cisplatin, an anticancer drug, forms DNA interstrand cross-links (ICL) that interfere with replication, whereas TREX2 is a 3'-->5' exonuclease that removes 3' mismatched nucleotides and promotes cellular proliferation. Here, we show that TREX2 is depleted in human cells derived from cancer after exposure to cisplatin but not other genotoxins including another cross-linking agent, mitomycin C (MMC), indicating a potential role for TREX2 depletion in cisplatin-induced cytotoxicity. To better understand TREX2 cellular function, we deleted TREX2 in mouse embryonic stem (ES) cells by gene targeting and find these cells exhibit reduced proliferation and gross chromosomal rearrangements including Robertsonian translocations (RbT). Quite interestingly, ES cells exposed to cisplatin also exhibit RbTs. By contrast, RbTs are not observed for ES cells exposed to MMC, indicating that RbTs are not caused by ICLs but instead TREX2 depletion by either cisplatin exposure or mutation. Taken together, our results show that cisplatin depletes TREX2 and causes genomic instability that is similarly observed in TREX2-mutant cells. Thus, cisplatin has two potential cytotoxic activities: (a) the generation of ICLs and (b) the depletion of TREX2.

LncRNA H19 promotes epithelial mesenchymal transition and metastasis of esophageal cancer via STAT3/EZH2 axis.

BACKGROUND: Long non-coding RNA H19 (lncRNA H19) has been widely reported in esophageal cancer (EC), and previous study had found that lncRNAH19 was up-regulated in EC and promoted cell proliferation and metastasis. However, the mechanism still needs further studied. METHODS: Levels of lncRNA H19 were analyzed by qRT-PCR in matched samples from 30 patients. Expression levels of lncRNA H19, let-7, STAT3 and EZH2 were additionally identified by qRT-PCR and western blotting in five EC cell lines. The effects of lncRNA H19 on cell proliferation, migration, invasion and apoptosis in cell lines were performed by MTT assay, colony formation assay, Transwell assay and flow cytometry in vitro, and tumor formation was detected by xenograft nude mice model in vivo. The expression level of STAT3, EZH2, ß-catenin, and EMT and metastasis related molecules such as E-cadherin, N-cadherin, Snail-1 and MMP-9 was assessed by qRT-PCR and western blotting. Finally, luciferase reporter assay and RIP assay were used to verify the interaction between lncRNA H19 and let-7c, and their subsequent regulation of STAT3. RESULTS: Knockdown of lncRNA H19 repressed cell proliferation, migration and invasion as well as EMT and metastasis via STAT3-EZH2-ß-catenin pathway, while lncRNA H19 regulated STAT3 negatively regulated let-7c in EC cell lines. CONCLUSIONS: lncRNA H19 facilitates EMT and metastasis of EC through let-7c/STAT3/EZH2/ß-catenin axis.

miR-126/VCAM-1 regulation by naringin suppresses cell growth of human non-small cell lung cancer.

Certain studies have indicated that naringin possesses various pharmacological activities including anti-aging, anti-oxidation, anticancer, cardiovascular and cerebrovascular disease prevention, in addition to anti-hepatic effects. The present study explores the anticancer effect of naringin on human small cell lung cancer H69AR cells. Cell growth and apoptosis rates of H69AR cells were measured by MTT or flow cytometry, which demonstrated naringin suppressed cell growth and induced apoptosis of H69AR cells. MicroRNA (miR)-126 expression and levels of phosphorylated protein kinase B (AKT), mechanistic target of rapamycin (mTOR), nuclear factor (NF)-κB and vascular cell adhesion molecule 1 (VCAM-1) proteins were detected by quantitative polymerase chain reaction and western blotting. It was identified that naringin increased miR-126 expression and suppressed the phosphorylation of AKT, mTOR, NF-κB and VCAM-1 proteins in H69AR cells. Suppression of miR-126 expression reduced the anticancer effects of naringin on H69AR cells, reversed the naringin-induced reduction of phosphoinositide 3-kinase/AKT/mTOR, and suppressed VCAM-1 protein levels. However, close of miR-126 expression did not affect the levels of NF-κB protein in H69AR cells. In summary, naringin exhibits its anti-cancer effect by suppressing cell growth of small cell lung cancer cells through miR-126/VCAM-1 signaling pathway.

Successful removal of giant mediastinal lipoma and liposarcoma involving both chest cavities: Two case reports.

RATIONALE: Surgical removal of a giant mediastinal lipoma or liposarcoma involving both chest cavities is always challenging. PATIENT CONCERNS: We present 2 cases of giant mediastinal tumor, one of which was a 22-year-old female who was admitted to our hospital due to a mild dyspnea after running. Computed tomography (CT) scan revealed a large mass with low density occupying the entire right hemithorax and extending anteriorly into the left. The other patient was a 43-year-old male, who was presented to the hospital with complaints of gradually progressive dyspnea. CT scan revealed a mass comprised of fat density with areas of soft-tissue density in-between, involving in both chest cavities, draping around the heart and great vessels. INTERVENTIONS: Both of the patients receive complete resection through a standard median sternotomy. DIAGNOSES: Histologic examination revealed lipoma for the first patient, and well differentiated liposarcoma for the second. OUTCOMES: Both of their symptoms were improved after surgery and the postoperative courses were good. LESSONS: Our experience indicated that complete surgical removal through a standard median sternotomy is a safe and efficient approach for the treatment of giant mediastinal lipoma and liposarcoma.

[Clinical application of video-assisted thoracoscopic surgery].

OBJECTIVE: To explore the clinical application of video-assisted thoracoscopic surgery (VATS). METHODS: We retrospectively analyzed the clinical data of 672 cases of VATS. There were 17 thoracic diseases such as emphysema, bullectomy for spontaneous pneumothorax, massive bullae, benign tumor of mediastinum, cyst of mediastinum, pulmonary benign tumors, hydropericardium, malignant pleural fluid, etc. RESULTS: The mean operation time was 57 minutes and there were no intraoperative complications. The bleeding during the operation was less than 100 mL. Postoperative pneumothorax occurred in 4 patients and among them 2 patients were of relapse after 1 month. The intrathoracic drain in most patients was removed with an average of 2. 5 days. A supplementary incision was needed in 10 cases: Six were due to the adhesion of full pleural cavity and 4 were found with the malignant tumor during the operation. CONCLUSION: VATS is an alternative approach that provides a safe, less invasive, and effective operation for treating spontaneous pneumothorax, benign tumor of mediastinum, cyst of mediastinum, pulmonary benign tumors, pericardial perfusion, and acute chest trauma patients.