Combination of oncolytic adenovirus ZD55 harboring TRAIL-IETD-MnSOD and cytokine-induced killer cells against lung cancer.

BACKGROUND: Our study aimed to investigate the effect of cancer-targeting gene-virotherapy and cytokine-induced killer (CIK) cell immunotherapy on lung cancer. METHODS: CIK cells were obtained from peripheral blood mononuclear cells using interferon (IFN)-γ, interleukin (IL)-2, and CD3 monoclonal antibody. The CIK cells were infected with oncolytic adenovirus ZD55 harboring tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), manganese-containing superoxide dismutase (MnSOD), and TRAIL-isoleucine-aspartate-threonine-glutamate (IETD)-MnSOD. The cells were then cocultured with lung cancer cell lines A549 and NCI-H1650, normal cell line BEAS-2B, or injected into an A549 xenograft mouse model. RESULTS: Proliferation, colony formation, and invasion of A549 and NCI-H1650 cells were significantly inhibited by co-cultivation with CIK cells carrying oncolytic adenoviruses (in order) ZD55-TRAIL-IETD-MnSOD > ZD55-TRAIL + ZD55-MnSOD > ZD55-MnSOD > ZD55-TRAIL. Compared to BEAS-2B cells, the production of IFN-γ, TNF-α, and lactate dehydrogenase (LDH) in tumor cells was increased. Tumor volume in the xenograft model and Ki-67 expression in tumor samples were reduced after injection of CIK cells carrying oncolytic adenoviruses, in the same order as the in vivo experiments. Levels of IFN-γ, TNF-α, and LDH contents were also increased in the same order. CONCLUSIONS: Our studies confirmed the high efficacy of combined oncolytic adenovirus ZD55 harboring TRAIL-IETD-MnSOD and CIK cells against lung cancer.

Primary epithelioid rhabdomyosarcoma of the stomach: a case report and review of literature.

BACKGROUND: Epithelioid rhabdomyosarcoma is a rare tumor that generally occurs in the bladder, the parotid gland, or the skin of the neck. We describe an unusual case of primary epithelioid rhabdomyosarcoma of the stomach and review the literature. CASE PRESENTATION: A 64-year-old woman presented with a lesion at the gastroesophageal junction. Histopathological examination showed irregularly sized round cells with low cytoplasmic content and eccentric nuclei. Mitotic figures were present. Fibrovascular septa and areas of necrosis were observed between tumor cells. Tumor cells were strongly positive for MyoD1, desmin, and myogenin, and weakly positive for actin, CD56, and PGP9.5. The ki-67 index was ≥90%. CONCLUSIONS: Primary epithelioid rhabdomyosarcoma of the stomach is extremely rare. Better awareness of this entity is necessary for early diagnosis and treatment.

Gastric aggressive fibromatosis: report of a case and review of the literature.

OBJECTIVES: To describe a rare case of aggressive fibromatosis of the stomach and discuss the differential diagnoses. METHODS: A 47-year-old man presented with nonspecific abdominal pain. Gastroscopy revealed stomach wall swelling. An antral gastrectomy was performed. Histological examination revealed spindle-shaped cells and morphology typical of aggressive fibromatosis. We performed a literature search to identify conditions with features similar to those of aggressive fibromatosis. RESULTS: Aggressive fibromatosis does not metastasize, but it is locally invasive and has a tendency to relapse; however, our patient has not had recurrence > 1 year after surgery. Aggressive fibromatosis of the stomach may be confused with an inflammatory fibroid polyp, a gastrointestinal stromal tumor, schwannoma, leiomyoma, inflammatory myofibroblastic tumor, scirrhous carcinoma of the stomach, follicular dendritic cell sarcoma, inflammatory malignant fibrous histiocytoma, myofibroma/myofibromatosis, and solitary fibrous tumor of the stomach. CONCLUSIONS: Aggressive fibromatosis of the stomach is a rare spindle cell tumor that must be differentiated from a variety of conditions.

LncRNA SOX21-AS1 is associated with progression of hepatocellular carcinoma and predicts prognosis through epigenetically silencing p21.

Long non-coding RNAs (lncRNAs) have been widely reported in various cancers due to their special molecular mechanisms. LncRNA SOX21-AS1 has been discovered to be a tumor facilitator in several types of human cancers. However, the expression pattern, clinical value and biological effects in hepatocellular carcinoma (HCC) are still unknown. In this study, we detected the high expression level of SOX21-AS1 in tumor tissues and cell lines through performing qRT-PCR analysis. The prognostic value of SOX21-AS1 was identified. Moreover, the biological effects of SOX21-AS1 on HCC cell activities were evaluated by functional assays, such as MTT, colony formation assay and transwell assay. As a result, silenced SOX21-AS1 suppressed cell proliferation and metastasis, resulted in cell cycle arrest, and induced apoptosis in hepatocellular carcinoma. Mechanically, RIP was conducted to prove that SOX21-AS1 could bind with EZH2. ChIp assay was carried out and manifested that SOX21-AS1 epigenetically silenced p21 via recruiting EZH2 to the promoter of p21. Finally, rescue assays were designed and carefully conducted to investigate whether SOX21-AS1 can interact with p21 to affect hepatocellular carcinoma progression. Generally, our results suggested that SOX21-AS1 could be a potential prognostic biomarker or a therapeutic target for hepatocellular carcinoma.

Microwave ablation versus hepatic resection for the treatment of hepatocellular carcinoma and oesophageal variceal bleeding in cirrhotic patients.

PURPOSE: The aim of this study was to compare the results of microwave ablation (MWA) and hepatic resection (HR) when combined with pericardial devascularisation plus splenectomy (PCDV) for the treatment of patients with cirrhosis complicated by small hepatocellular carcinoma (HCC) and oesophageal variceal bleeding (EVB). MATERIALS AND METHODS: Between 2001 and 2013, 73 patients (median age 53.2 years, 67% male) with small HCC and concomitant EVB who underwent MWA or HR for HCC and PCDV for cirrhotic portal hypertension were selected retrospectively for inclusion in this study. The overall survival curves and recurrence-free survival curves were calculated using the Kaplan-Meier method and compared using log-rank tests. Multivariate analysis was performed using the Cox regression model. RESULTS: The 1-, 3- and 5-year overall survival rates were 95.2%, 71.4% and 38.1% and 96.7%, 53.3% and 43.3% for the HR and MWA groups, respectively; these did not differ significantly between the two groups. However, patients in the HR group had more post-operative complications (52.3% vs. 13.7%; p = 0.002). Multivariate analysis identified albumin and bilirubin levels and tumour size to be statistically significant and independent prognostic factors for overall survival, while BCLC stage was associated with poor recurrence-free survival. Furthermore, albumin levels were shown to be an independent predictive factor for post-operative complications. CONCLUSIONS: For patients with small HCC and concomitant EVB, MWA plus PCDV may reduce the incidence of post-operative complications relative to and provide similar therapeutic benefits as HR plus PCDV, especially for patients with low albumin levels.

High-mobility group protein box1 expression correlates with peritumoral macrophage infiltration and unfavorable prognosis in patients with hepatocellular carcinoma and cirrhosis.

BACKGROUND: High-mobility group protein box1 (HMGB1) is a pivotal factor in the development and progression of many types of tumor. Its role in hepatocellular carcinoma (HCC), and especially its correlation with intratumoral and peritumoral macrophage infiltration, remains obscure. We analyzed the potential roles and prognostic value of HMGB1 and explored the correlation between HMGB1 and macrophage infiltration in HCC using clinical samples. METHODS: We reviewed clinicopathological and follow-up data on a cohort of 149 patients with HCC complicated with Hepatitis B-related cirrhosis. We measured the expression of HMGB1 and CD68 in tumoral and peritumoral liver tissues after curative resection and assessed the impacts of the tumor-associated macrophage (TAM) count and HMGB1 expression on clinicopathologic characteristics, overall survival (OS), and recurrence-free survival (RFS). RESULTS: Ninety-four of the patients had elevated tumoral HMGB1 expression and 59 of the patients had elevated peritumoral HMGB1 expression, compared to only 4 patients with elevated peritumoral HMGB1 expression in 36 pateints with Hepatitis B virus (HBV)-negative HCC without liver cirrhosis (p < 0.001). The peritumoral HMGB1 expression levels were correlated with tumor invasiveness, BCLC stage, and recurrence. The degree of TAM infiltration was higher in peritumoral tissues with high HMGB1 expression than in peritumoral tissues with low HMGB1 expression (p < 0.001). There was no significant difference in TAM infiltration between tumoral tissues with high and low HMGB1 expression. Kaplan-Meier analysis showed that intratumoral HMGB1 overexpression was associated with poor OS, but not with RFS. High peritumoral HMGB1expression and TAM count, which correlated positively with tumor size and BCLC stage, were independent prognostic factors for OS (p < 0.001 and p = 0.017, respectively) and RFS (p = 0.002 and p = 0.024, respectively). Multivariate analyses indicated peritumoral HMGB1 expression (p = 0.014) and TAM count (p = 0.037), as well as tumor differentiation (p = 0.026), to be independent significant prognostic factors for RFS. CONCLUSIONS: High HMGB1 expression in peritumoral liver tissues correlated with peritumoral macrophage infiltration and had prognostic value in HCC, suggesting that peritumoral HMGB1 might show promise as a new biomarker to predict HCC progression.

The preclinical evaluation of TIC10/ONC201 as an anti-pancreatic cancer agent.

Here we evaluated the potential anti-pancreatic cancer activity by TIC10/ONC201, a first-in-class small-molecule inducer of tumor necrosis (TNF)-related apoptosis-inducing ligand (TRAIL). The in vitro results showed that TIC10 induced potent cytotoxic and cytostatic activities in several human pancreatic cancer cell lines (Panc-1, Mia-PaCa2, AsPC-1 or L3.6). TIC10 activated both extrinsic (TRAIL-caspase-8-dependent) and endogenous/mitochondrial (caspase-9-dependent) apoptosis pathways in the pancreatic cancer cells. Molecularly, we showed that TIC10 inhibited Akt-Erk activation, yet induced TRAIL expression in pancreatic cancer cells. Significantly, TIC10, at a relatively low concentration, sensitized gemcitabine-induced growth inhibition and apoptosis against pancreatic cancer cells. Further, TIC10 and gemcitabine synergistically inhibited Panc-1 xenograft growth in SCID mice. The combination treatment also significantly improved mice survival. In addition, Akt-Erk in-activation and TRAIL/cleaved-caspase-8 induction were observed in TIC10-treated Panc-1 xenografts. Together, the preclinical results of the study demonstrate the potent anti-pancreatic cancer activity by TIC10, or with gemcitabine.

Combined Endovascular Embolization and Open Surgery for Splenic Artery Aneurysm with Arteriovenous Fistula.

Splenic artery aneurysm with arteriovenous fistula is extremely rare; however, it is clinically important because of the potential of aneurysm rupture and gastroesophageal variceal hemorrhage. Most previous cases were managed by surgery directly. We present a case which was successfully treated with combined endovascular embolization and open surgery. It may be a safe and effective approach to manage this entity.

Effect of Liuweidihuang pill and Jinkuishenqi pill on inhibition of spontaneous breast carcinoma growth in mice.

OBJECTIVE: To investigate the preventing and treating action of Liuweidihuang pill (LP) and Jinkuishenqi pill (JP) on spontaneous breast carcinoma in mice. METHODS: A model of spontaneous breast carcinoma was derived from 11.5-month-old female Kunming breeding mice following the delivery of several litters. The mice were randomly divided into five groups: model control group (C), Liuweidihuang pill high-dose group (LH; 4.6 g · kg(-1) · d(-1)), Liuweidihuang pill low-dose group (LL; 2.3 g · kg(-1) · d(-1)), Jinkuishenqi pill high-dose group (JH; 4.6 g · kg(-1) · d(-1)) and Jinkuishenqi pill low-dose group (JL; 2.3 g · kg(-1) · d(-1)). Cancer tissue volume was measured by water immersion. Histopathology was analyzed by hematoxylin and eosin staining. Vascular endothelial growth factor (VEGF), extracellular signal-regulated kinase (ERK) and cyclin D1 protein expression in cancer tissue was assayed by western blotting. RESULTS: Compared with the control group, cancer tissue volume and weight were lower in the LP and JP groups, and survival time was longer. The expression of VEGF, ERK and Cyclin D1 were inhibited in the LP and JP groups (P < 0.05), and cell differentiation was increased. Tumor weights and volumes and VEGF, ERK and Cyclin D1 expression in LL or LH were significantly lower than in JL and JH (P < 0.01). CONCLUSION: Both LP and JP could restrain cancer growth and promote cancer cell differentiation; moreover, LP was more effective than JP The likely mechanism of action was via inhibition of VEGF, ERK and cyclin D1.

Submucosal Lipoma: a Rare Cause of Recurrent Intestinal Obstruction and Intestinal Intussusception.

Intestinal lipomas are rare nonepithelial tumors that are usually detected incidentally. They are usually asymptomatic, but lipomas larger than 2 cm may become symptomatic due to obstruction, bleeding, or intussusception. Adult intussusception due to intestinal lipoma is a very rare condition. In this paper, we report a case of small bowel lipoma that became symptomatic due to intermittent obstruction episodes and ileo-ileal intussuception. Segmental ileal resection was performed, and histopathological examination of the resected specimen confirmed the diagnosis of lipoma.

A model of spontaneous mouse mammary tumor for human estrogen receptor- and progesterone receptor-negative breast cancer.

Breast cancer (BC) is the most frequently malignancy in women. Therefore, establishment of an animal model for the development of preventative measures and effective treatment for tumors is required. A novel heterogeneous spontaneous mammary tumor animal model of Kunming mice was generated. The purpose of this study was to characterize the spontaneous mammary tumor model. Histopathologically, invasive nodular masses of pleomorphic tubular neoplastic epithelial cells invaded fibro-vascular stroma, adjacent dermis and muscle tissue. Metastatic spread through blood vessel into liver and lungs was observed by hematoxylin eosin staining. No estrogen receptor (ER) or progesterone receptor (PR) immunoreactivity was detected in their associated malignant tumors, human epidermal growth factor receptor-2 (HER-2) protein weak expression was found by immunohistochemistry. High expression of vascular endothelial growth factor (VEGF), moderate or high expression of c-Myc and cyclin D1 were observed in tumor sections at different stages (2, 4, 6 and 8 weeks after cancer being found) when compared with that of the normal mammary glands. The result showed that the model is of an invasive ductal carcinoma. Remarkably in the mouse model, ER and PR-negative and HER2 weak positivity are observed. The high or moderate expressions of breast cancer markers (VEGF, c-Myc and cyclin D1) in mammary cancer tissue change at different stages. To our knowledge, this is the first report of a spontaneous mammary model displaying colony-strain, outbred mice. This model will be an attractive tool to understand the biology of anti-hormonal breast cancer in women.

The morphometrical analysis on the ultrastructure of A549 cells.

OBJECTIVE: To report the morphometric characteristics of ultrastructure inside A549 cells. METHODS: A549 cells were processed for inverted microscopy and transmission electron microscopy (TEM). Cell images were obtained randomly using inverted microscopy and TEM. The morphometric parameters of ultrastructure were tested using precise morphometric techniques by Image-Pro Plus analysis software. RESULTS: (1) The diameter of A549 cells from inverted microscopy and TEM images was 14.93 µm and 10.59 µm. (2) By defining cell as reference space the volume densities (VV) of nucleus and cytoplasm were about 0.28 and 0.72; the surface densities (SV) of nucleus were 0.19 µm-1. By defining cell nucleus as reference space the VV of nucleoli, euchromatin and heterochromatin were 0.076, 0.72 and 0.20 respectively; the SV of nucleoli was 0.15 µm-1. By defining cytoplasm as reference space the VV of mitochondria, lamellar bodies and lysosomes were 0.046, 0.025 and 0.014; the SV of mitochondria, lamellar bodies and lysosomes were 0.60 µm-1, 0.36 µm-1, and 0.18 µm-1. (3) In individual A549 cell total volume and surface of mitochondria were 61.91 µm³ and 1001.67 µm²; Total volume and surface area of lamellar bodies were 76.82 µm³ and 428.68 µm²; Total volume and surface area of lysosomes were 21.69 µm³ and 212.04 µm². CONCLUSIONS: The morphometric parameters of some ultrastructures within A549 cells were established using precise morphometric techniques by Image-Pro Plus analysis software.

[Comparative genomic hybridization analysis of nasopharyngeal carcinoma drug-resistant cell CNE2/DDP and its parent cell CNE2].

BACKGROUND & OBJECTIVE: Developed from fluorescence in situ hybridization (FISH), comparative genomic hybridization (CGH) is a new molecular and cellular technique,used to examine the genomic imbalances,especially the loss and amplification of chromosomes,and to locate these alterations in certain chromosome regions. For a comprehensive understanding of the possible differences in genomic DNA between nasopharyngeal carcinoma drug-resistant and drug-sensitive cells, we analyzed the genomic DNA of a nasopharyngeal carcinoma drug-resistant cell line CNE2/DDP and its parent cell line CNE2 with CGH. METHODS: Genomic DNA was extracted from CNE2/DDP and CNE2 cells, as well as from normal placenta tissue. Fluorescent random primed labeling method was used to label the DNA probes (CNE2 and CNE2/DDP cells with green fluorescein-12-dUTP and normal placenta tissue with red tetramethylrhodamine-5-dUTP). The labeled DNA probes were then co-hybridized with normal lymphocyte metaphase chromosomes. Signals were taken by charge coupled device (CCD) and processed with Quips CGH Program after fluorescent hybridization. The green-to-red fluorescent ratio was calculated automatically and showed with graphs. RESULTS: There were extensive chromosome changes in CNE2, mainly the gain of 1q, 3q, 5p, 6p, 7p, 8q, 9q, 11p, 12q and 19q, and the loss of 4q, 12p, 13p, 14p, 15p, 18, 20q, 21p and 22. However, the CNE2/DDP cells, which come from the CNE2, showed a relatively normal karyotype except loss of 8p and gain of 8q and 19q. Consistent result was achieved after the CNE2/DDP cells were cultured in the medium free of any drugs for over one month. It appears that the CNE2/DDP cell has relative normal and much more stable chromosome constitution than its parent CNE2 cell. CONCLUSION: The CNE2/DDP is a single drug-resistant clone selected from CNE2,which is a blend of sub-clones that have different sorts of chromosome number and structure. It strongly suggests that the appearance of tumor drug-resistance is mainly a select process in which the would-be drug-resistant cell clone be selected from unfavorable survival condition.

[Establishment of a human nasopharyngeal carcinoma drug-resistant cell line CNE2/DDP and screening of drug-resistant genes].

BACKGROUND & OBJECTIVE: Chemotherapy constitutes one of the chief supplementary methods in the treatment of nasopharyngeal carcinoma (NPC). However, the appearance of drug resistance often causes failure of chemotherapy. For overcoming drug resistance, it is of great importance to screen drug-resistant associated genes so as to identify potential molecular targets. This study was designed to establish a drug-resistant cell line from a human nasopharyngeal carcinoma cell line CNE2, and to screen human nasopharyngeal carcinoma drug-resistant genes by a new strategy based on improved subtractive hybridization. METHODS: The drug-resistant cell line was established by a program of treating the human nasopharyngeal carcinoma cells CNE2 in the medium with repeated sharp high and then low but gradually increasing concentration of cisplatin. Drug sensitivity was measured by MTT assay. Fluorescence activated cell analysis(FACS) was employed for determining the concentration of fluorescence dye rhodamine 123 within the cells. Cell growth curve, doubling time, and cell morphology were measured and observed. The drug-resistant genes were screened by a new strategy of PCR-based subtractive hybridization. Sequencing and blast analysis were performed after the differentially expressed genes had been verified by reverse dot blotting. The result was further confirmed by RT-PCR. RESULTS: The resistance indexes of CNE2/DDP to cisplatin (DDP), 5-fluorouracil (5-FU), and vincristine (VCR) were 27.9, 227.9, and 55.5, respectively, indicating its multi-drug resistant property. FACS analysis showed that the concentration of rhodamine 123 was much lower in CNE2/DDP cells than in CNE2 cells (12.98 vs. 243.62). The CNE2/DDP cells appeared smaller, more regularly round, and longer doubling time (26 hours vs. 19 hours) than CNE2 cells. Six differentially expressed sequences were discovered using improved subtractive hybridization; all of them were found to be homologous to known genes after sequencing analysis. Three of them were highly expressed in CNE2/DDP cells. Among them, one sequence, which encodes a 79 amino acid protein,known as DC13 protein (DC13), was a function unknown gene which has certain relationship with malignancy. The other two sequences were ubiquitin C gene and NADH dehydrogenase subunit 2 (ND2) gene, respectively. The other three of the six sequences, whose expression were inhibited in CNE2/DDP cells, were cytochrome C oxidase subunit I(COX1), ribosomal protein L27(RPL27),and ribosomal protein S27 (RPS27) genes, respectively. CONCLUSION: A drug- resistant cell line CNE2/DDP, which showed a typical resistant phenotype to anti-cancer drugs was established. The PCR-based improved subtractive hybridization is an effective approach to identify differentially expressed genes. Many genes, both known and unknown, might contribute to the existence of drug-resistant phenotype, through increasing or decreasing their expression.