Distant Lung Metastasis Research Articles

The results of surgical treatment in patients with liver alveococcosis in a hepato-pancreato-biliary center (a 10-years’ experience)

Background: Aggressive course of liver alveococcosis makes it possible to designate it as a “parasitic liver cancer”. The main treatment method for the disease is surgery. The parasitic mass is resected according to R0 principles, with concomitant plastic surgery of the major vessels and bile ducts to increase resectability.Aim: To assess the potential of surgical treatment in patients with advanced liver alveococcosis using transplantation techniques.Materials and methods: We retrospectively analyzed in- and outpatient medical files of 62 subjects with confirmed liver alveococcosis, who had been treated in the Volga District Medical Centre (Nizhny Novgorod, Russia) from 2008 to 2018. Thirty two (32) patients had advanced liver alveococcosis with involvement of afferent and efferent vasculature and biliary tract. Surgical procedures were used in 50/62 patients (or 4.2% of the total number of liver resections performed during this time interval, n = 1197). Complications occurred in 46% (23 / 50) of the cases. Twenty nine (29, or 58%) patients had been operated before (mostly cytoreductive resections and/or explorative laparotomies). Distant lung metastases were found in 2 (4%) patients.Results: Fifty (50) patients had curative surgical procedures: liver resections in 45, deceased donor orthotopic liver transplantations in 5. Most common were extensive liver resections (more than 4 segments). Resection and reconstruction of the main vessels were necessary in 50% (25 cases) of the patients, including v. cava inferior in 25 cases and the portal vein in 24 cases. In 31 patients, resection and reconstruction of extra-hepatic bile ducts was performed, and in 17 (33%) patients resections of the neighboring organs, such as diaphragm, lung, right adrenal, duodenum, stomach, and colon. In 4 cases, resections were performed ex situ ex vivo, followed by auto-transplantation, including 2 cases with reverse auto-transplantation of the left lateral sector to the right. The incidence of liver failure events grade A and B (by International Study Group of Liver Surgery, ISGLS) did not exceed 10% (4 patients). Complications were seen in 25 cases, including Clavien – Dindo Grade II in 5, Grade IIIb in 13, Grade IVb in 2, and Grade V in 5. The number of bile leakage events (ISGLS) class B was 6 and class C 10. All patients underwent obligatory adjuvant anti-parasitic therapy.Conclusion: At present, surgical treatment of liver alveococcosis remains a method of choice, that requires that the hepato-pancreato-biliary center would have in place a well-developed transplantation program, adequate equipment and well-trained surgical and anesthetic teams.

Upregulation of lncRNA ADAMTS9-AS2 Promotes Salivary Adenoid Cystic Carcinoma Metastasis via PI3K/Akt and MEK/Erk Signaling.

Neurotropic infiltrative growth and distant metastasis are the main causes of death in salivary adenoid cystic carcinoma (SACC) patients. Long noncoding RNAs (lncRNAs) are involved in many human neoplasms, however, their potential roles in SACC are unclear. In our study, we found that ADAM metallopeptidase with thrombospondin type 1 motif, 9 (ADAMTS9) antisense RNA 2 (ADAMTS9-AS2) was significantly upregulated in SACC patients with metastasis and SACC-lung metastasis (LM) cells. Moreover, ADAMTS9-AS2expression was closely associated with the prognosis anddistant metastasis in SACC patients. Next, we found thatc-myc could specifically bind to the promoter of ADAMTS9-AS2 and activated its transcription. Knockdown of ADAMTS9-AS2 significantly inhibited migration and invasion of SACC cells invitro and distant lung metastasis invivo. Furthermore, ADAMTS9-AS2, which mainly expressed in the cytoplasm, shared microRNA (miRNA) response elements with Integrin α6 (ITGA6). Overexpression of ADAMTS9-AS2 competitively bound to miR-143-3p that inhibited ITGA6 from miRNA-mediated degradation, and thus it activated the activity of PI3K/Akt and MEK/Erk signaling and facilitated SACC metastasis. In summary, ADAMTS9-AS2 promotes migration and invasion in SACC by competing with miR-143-3p. This sheds a new insight into the regulation mechanism of ADAMTS9-AS2, and it provides a possible application for the SACC treatment.

Bioinspired Hybrid Protein Oxygen Nanocarrier Amplified Photodynamic Therapy for Eliciting Anti-tumor Immunity and Abscopal Effect.

An ideal cancer therapeutic strategy is expected to possess potent ability to not only ablate primary tumors but also prevent distance metastasis and relapse. In this study, human serum albumin was hybridized with hemoglobin by intermolecular disulfide bonds to develop a hybrid protein oxygen nanocarrier with chlorine e6 encapsulated (C@HPOC) for oxygen self-sufficient photodynamic therapy (PDT). C@HPOC realized the tumor-targeted co-delivery of photosensitizer and oxygen, which remarkably relieved tumor hypoxia. C@HPOC was favorable for more efficient PDT and enhanced infiltration of CD8+ T cells in tumors. Moreover, oxygen-boosted PDT of C@HPOC induced immunogenic cell death, with the release of danger-associated molecular patterns to activate dendritic cells, T lymphocytes, and natural killer cells in vivo. Notably, C@HPOC-mediated immunogenic PDT could destroy primary tumors and effectively suppress distant tumors and lung metastasis in a metastatic triple-negative breast cancer model by evoking systemic anti-tumor immunity. This study provides a paradigm of oxygen-augmented immunogenic PDT for metastatic cancer treatment.

PAPILLARY THYROID CARCINOMA IN CHILDREN AND ADOLESCENTS, RESULTS OF TREATMENT OF 215 PATIENTS: FACTORS PREDICTING DISTANT METASTASES AND RECURRENCE OF THE DISEASE

We evaluated the results of treatment of 215 pediatric patients with papillary thyroid carcinoma (PTC). Median age at diagnosis was 14 (5 - 18 y.o). Thyroidectomy performed in 105, subtotalthyroidectomy in 8, lobectomy - in 102, additional lymph node dissection - in 169, radioiodine ablation/ therapy - in 102 patients. Follicular variant PTC was found in 41 (19%), solid - in 37 (17%), diffuse sclerosing - in 3 (1%), the remaining had a classical PTC with mainly papillary (90; 42%) or follicular (44; 21%) pattern. 21% had multifocal PTC, 4,6% - extrathyroidal extension, 62% - regional lymph node metastases, 12% - lung metastases. The follow up was 95 months (6 - 301 mo). All patients are alive. Recurrence of PTC occurred in 37 (17%): 7 - thyroid remnant, 26 - cervical lymph nodes, 4 - distant lung metastases. Prognostic factors for distant metastases were young age at diagnosis <12 years old (OR 3.4, р <0.022), tumor size more then 3 cm (OR 3.2, р <0.027) and lateral cervical lymph node metastases (OR 21.39, р <0.0001). Risk of recurrence was higher in multifocal PTC (OR 2.4, р <0.009), regional lymph node metastases (OR 7.4, р <0,008) and distant metastases (OR 3.2, р <0,01). The highest risk of relapse had the patients with solid variant of PTC. Radical surgery diminished the risk of relapse (OR 0.3, р <0.002). There was no evidence of impact of radioiodine therapy on risk of recurrence (р=0.66).

Distant lung metastases caused by a histologically benign phosphaturic mesenchymal tumor.

SummaryTumor-induced osteomalacia (TIO) is a rare form of hypophosphatemia usually caused by phosphaturic mesenchymal tumors (PMTs); the biologic behavior of PMTs is under investigation. Herein we present a case of TIO with a protracted course over 12 years leading to a fatal outcome. A 39-year-old man presented with weakness in 2004 and was found to have decreased serum phosphorus, phosphaturia and low levels of 1,25-dihydroxyvitamin D3. Four years later he developed a painful left calf mass. The lesion was resected, but recurred causing extreme pain and dysfunction. Radiological examination showed a large cluster of soft tissue tumors affecting all the muscle compartments of the calf and a smaller lesion inside the metaphysis of the tibia. Above-knee amputation was performed. Histological examination of all lesions showed a cellular spindle cell neoplasm with variously sized vessels, wide vessel-like spaces and scattered deposits of calcified extracellular material. The tumor infiltrated skeletal muscles, subcutaneous fat and the proximal end of the fibula. The tibial lesion had identical histology. Three years after the amputation the patient presented with cough and dyspnea. Radiological examination, followed by an open biopsy, showed that there were multiple metastatic nodules of PMTs in both lungs. Shortly after the diagnosis the patient died. This case illustrates that even benign cases of PMTs may lead to a fatal outcome and the classification of PMTs into benign and malignant should be reassessed in order to correspond to its biological behavior.Learning points:PMTs, aside from having locally aggressive behavior, may metastasize and cause deathPMTs may behave aggressively despite ‘benign’ histological findings Accurate diagnosis of tumor-induced osteomalacia and patient management require a multidisciplinary approach

Cell-penetrating peptide-based nanovehicles potentiate lymph metastasis targeting and deep penetration for anti-metastasis therapy.

Lymph metastasis is a vital pathway of cancer cell dissemination, and insidious lymph node metastasis increases the risk of distant cancer metastasis. Current therapies for lymph metastasis are largely restricted by limited targeting and penetration capacity. Herein, we report that an r9 cell-penetrating peptide-based cabazitaxel nanovehicle (r9-CN) displays prominent lymph metastasis targeting and deep penetration ability after intravenous injection for effective anti-metastasis therapy.Methods: The r9-CN and CN nanovehicles were prepared by thin film dispersion, using DSPE-PEG2000 as the nano-carrier material and cabazitaxel as the model drug to fabricate r9-modified nano-micelles by self-assembly. The morphology, size, and stability in physiological solutions of r9-CN and CN were characterized. The targeting, biodistribution, deep penetration, and therapeutic efficacy of r9-CN and CN were systematically explored in vitro and in vivo.Results: The r9-CN nanovehicle consists of homogeneous particles with a mean diameter of 13 nm and zeta potential of +0.75 mV. Compared with the nanovehicle lacking the r9 peptide (CN), r9-CN exhibits long retention and deep penetration in the tumor mass, and considerably enhances accumulation and flexible permeation in metastatic lymph nodes, thereby notably suppressing primary tumor growth, lymph node metastasis, and distant lung metastasis.Conclusion: The cumulative findings reveal that r9-CN offers a promising delivery platform, enabling efficient lymph metastasis targeting and deep penetration for effective anti-metastasis therapy.

Compound Fuling Granule Suppresses Ovarian Cancer Development and Progression by disrupting mitochondrial function, galactose and fatty acid metabolism

Our previous studies have demonstrated that the compound fuling granule (CFG), a traditional Chinese medicine, suppresses ovarian cancer cell growth, migration and metastasis. However, the underlying mechanisms remain to be fully elucidated. In this study, we found that CFG could induce mitochondrial fragmentation, mitochondrial membrane potential reduction and cytochrome c release in ovarian SKOV3 cancer cells. In addition, both metabolomics and transcriptomics approaches were applied to illustrate the systemic mechanism of CFG on ovarian cancer formation and progression. To this end, we established two tumor-bearing mice models with subcutaneous injection or tail intravenous injection. Functionally, administration of CFG suppresses in situ tumor growth and distant lung metastasis. Subsequently, gas chromatography-mass spectrometry (GC-MS) was applied to determine the metabolic alterations among the plasma samples from these in vivo models. In the subcutaneous injection model, 26 distinguishable metabolites were identified and 12 metabolic pathways were reprogrammed. Meanwhile, 19 metabolites involved in 7 metabolic pathways showed significant differences in the tail intravenous injection model. Importantly, integrative metabolomics and transcriptomics analysis showed these metabolites were highly associated with galactose metabolism and fatty acid metabolism. This study suggests that CFG may suppress ovarian cancer cell proliferation and metastasis by regulating mitochondrion-related energy metabolisms.

STAT3-mediated upregulation of lncRNA HOXD-AS1 as a ceRNA facilitates liver cancer metastasis by regulating SOX4

BackgroundSeveral of the thousands of human long noncoding RNAs (lncRNAs) have been functionally characterized, yet their potential involvement in hepatocellular carcinoma (HCC) remains poorly understood.MethodsLncRNA-HOXD-AS1 was identified by microarray and validated by real-time PCR. The clinicopathological significance of HOXD-AS1 was analyzed by Kaplan-Meier method. Chromatin immunoprecipitation was conducted to examine the mechanism of HOXD-AS1 upregulation. The role of HOXD-AS1 in HCC cells was assessed both in vitro and in vivo. ceRNA function of HOXD-AS1 was evaluated by RNA immunoprecipitation and biotin-coupled miRNA pull down assays.ResultsIn this study, we found that HOXD-AS1 was significantly upregulated in HCC tissues. Clinical investigation demonstrated high expression level of HOXD-AS1 was associated with poor prognosis and high tumor node metastasis stage of HCC patients, and was an independent risk factor for survival. Moreover, our results revealed that STAT3 could specifically interact with the promoter of HOXD-AS1 and activate HOXD-AS1 transcription. Knockdown of HOXD-AS1 significantly inhibited migration and invasion of HCC cells in vitro and distant lung metastasis in vivo. Additionally, HOXD-AS1 was enriched in the cytoplasm, and shared miRNA response elements with SOX4. Overexpression of HOXD-AS1 competitively bound to miR-130a-3p that prevented SOX4 from miRNA-mediated degradation, thus activated the expression of EZH2 and MMP2 and facilitated HCC metastasis.ConclusionsIn summary, HOXD-AS1 is a prognostic marker for HCC patients and it may play a pro-metastatic role in hepatocarcinogenesis.

Abstract 22: The Y537S ESR1 mutation is a dominant driver of distant ER-positive breast cancer metastasis

Abstract Background: Estrogen receptor (ESR1) mutations occur at a high frequency in metastatic breast tumors in patients treated with hormonal therapy in the metastatic setting. We do not know if these mutations are involved in metastasis. Experimental design and methods: We generated ESR1 Y537S homozygous mutations using CRISPR Casp-9 technology. Treatment synergy was evaluated using Compusyn. Athymic mice were used in tumor xenograft studies. ChIP-Seq and transcriptome analyses were performed. Results: We generated CRISPR ESR1 Y537S mutation homozygous knock-in clones and lentiviral stable pools in MCF-7 cells. Transcriptome profiling revealed elevated expression of Hallmark pathways, including epithelial mesenchymal transition (EMT) and estrogen-regulated gene expression. Mutant cell growth was resistant to tamoxifen, but responsive to fulvestrant treatment. Synergistic treatment effects were observed with fulvestrant and the mTOR inhibitor everolimus or the CDK4/6 inhibitor palbociclib. CRISPR Y537S mutant knock-in cells grown in the mammary fat-pad of athymic mice spontaneously metastasized to distant organs including the lung, intestine, and kidneys. In the presence of estrogen, there was no difference in the frequency of distant macrometastases between parental wild-type ER and CRISPR Y537S mutant ER mice. However, in the absence of estrogen, mimicking aromatase inhibitor treatment, 80% of CRISPR Y537S mutant ER mice displayed overt distant macrometastases, but none were observed in parental wild-type ER mice (p=0.04). Distant tumors retained ER expression and hormone sensitivity. Comparison of residual tamoxifen-treated metastatic tumors with tumors grown at the primary mammary fat-pad site using immunoblot analysis demonstrated significant reduction in estrogen-regulated gene expression, but no effect on the expression of biomarkers associated with EMT, suggesting a disconnect between EMT and distant metastasis in mutant cells. EMT genes were also identified as direct binding site targets in Y537S mutant cells compared with wide-type ER using ChIPSeq. We discovered that expression of the Y537S mutant was dominant, driving the growth of distant metastatic tumors when co-expressed with wild-type ER cells. A Y537S ER mutant-specific gene expression signature predicted poor disease-free survival of ER-positive patients using the METABRIC database, and lung-specific metastasis-free survival in a Memorial Sloan Kettering dataset. Conclusion: The Y537S ER mutation is a driver of distant metastasis in ER-positive breast cancer cells. A Y537S ER mutant-specific gene expression signature predicted poor disease-free, and distant lung metastasis in ER-positive patients. Mutation status is a potential new predictive factor for hormone therapy of metastatic breast cancer patients on maintenance hormonal therapy. Citation Format: Guowei Gu, Lin Tian, Meng Gao, Yassine Rechoum, Luca Gelsomino, Derek Dustin, Arnoldo Corona-Rodriguez, Amanda R. Beyer, Anna Tsimelzon, Xiang Zhang, Sebastiano Ando', Suzanne Fuqua. The Y537S ESR1 mutation is a dominant driver of distant ER-positive breast cancer metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 22.

Abstract S4-02: The Y537S ESR1 mutation is a dominant driver of distant ER-positive breast cancer metastasis

Abstract Background: Estrogen receptor (ESR1) mutations occur at a high frequency in metastatic breast tumors in patients treated with hormonal therapy in the metastatic setting. We do not know if these mutations are involved in metastasis. Experimental design and methods: We generated ESR1 Y537S homozygous mutations using CRISPR Casp-9 technology. Treatment synergy was evaluated using Compusyn. Athymic mice were used in tumor xenograft studies. ChIP-Seq and transcriptome analyses were performed. Results: We generated CRISPR ESR1 Y537S mutation homozygous knock-in clones and lentiviral stable pools in MCF-7 cells. Transcriptome profiling revealed elevated expression of Hallmark pathways, including EMT and estrogen-regulated gene expression. The EMT in mutant cells was associated with a switch from E-cadherin to vimentin, and increased expression of SNAIL and TWIST. Mutant cell growth was resistant to tamoxifen, but responsive to fulvestrant treatment. Synergistic treatment effects were observed with fulvestrant and everolimus or palbociclib. CRISPR Y537S mutant knock-in cells grown in the mammary fat-pad of athymic mice spontaneously metastasized to distant organs including the lung, intestine, and kidneys. In the presence of estrogen, there was no difference in the frequency of distant macrometastases between parental wild-type ER and CRISPR Y537S mutant ER mice. However, in the absence of estrogen, 80% of CRISPR Y537S mutant ER mice displayed overt distant macrometastases, but none were observed in parental wild-type ER mice (p=0.04). Interestingly, although CRISPR Y537S mutant ER tumors grown in the mammary fat-pad were unresponsive to tamoxifen treatment, tamoxifen significantly inhibited the growth of mutant tumors at the distant microenvironment (8-fold). Distant tumors retained ER expression and hormone sensitivity. Comparison of residual tamoxifen-treated metastatic tumors with tumors grown at the primary mammary fat-pad site using immunoblot analysis demonstrated significant reduction in estrogen-regulated gene expression, but no effect on the expression of biomarkers associated with EMT, suggesting a disconnect between EMT and distant metastasis in mutant cells. EMT genes were also identified as direct binding site targets in Y537S mutant cells compared with wide-type ER using ChIPSeq. We discovered that expression of the Y537S mutant was dominant, driving the growth of distant metastatic tumors when co-expressed with wild-type ER cells. A Y537S ER mutant-specific gene expression signature predicted poor disease-free survival of ER-positive patients using the METABRIC database, and lung-specific metastasis-free survival in a Memorial Sloan Kettering dataset. Conclusion: The Y537S ER mutation is a driver of distant metastasis in ER-positive breast cancer cells. Although tamoxifen treatment was ineffective at reducing the growth of mutant cells grown at the primary site, it was effective at reducing distant metastasis. A Y537S ER mutant-specific gene expression signature predicted poor disease-free, and distant lung metastasis in ER-positive patients. Mutation status is a potential new predictive factor for hormone therapy of metastatic breast cancer patients on maintenance hormonal therapy. Citation Format: Fuqua SAW, Gu G, Rechoum Y, Gelsomino L, Dustin DJ, Corona-Rodriguez A, Beyer AR, Pejerrey SM, Gao M, Tsimelzon A, Tian L, Zhang X, Nagi C, Ando' S. The Y537S ESR1 mutation is a dominant driver of distant ER-positive breast cancer metastasis [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr S4-02.

State of lipid peroxidation and endogenous intoxication under non-small-cell lung cancer

Background: The problems of optimizing the methods of diagnosis and predicting the course of lung cancer occupying a leading position in the structure of cancer are still relevant. Aims and Objectives: To establish the patterns of changes in the parameters of endogenous intoxication and lipid peroxidation processes in the saliva of patients with non-small-cell lung cancer (NSCLC) depending on the histological type of a tumor. Materials and Methods: A total of 505 people took part in the case–control study: The main group (NSCLC, n = 290) and the control one (healthy, n = 215). All participants were questioned and underwent the biochemical examination of saliva and histological verification of the diagnosis. The parameters of endogenous intoxication and lipid peroxidation were determined spectrophotometrically. Results: The results of the study support the hypothesis of association of endogenous intoxication and lipid peroxidation processes with the development of NSCLC. It was shown that against the background of NSCLC; the level of primary lipid peroxidation products decreases. Thereat, the content of triene conjugates and Schiff bases, as well as the MM 280/254 nm distribution coefficient, increases. The growth of these indicators with an increase in the size of the tumor was noted. The differences due to the histological type of NSCLC have been identified. Conclusion: Thus, with squamous cell NSCLC, a higher level of secondary lipid peroxidation products was noted. The presence of distant metastases in lungs significantly contributes to changes in indicators of endogenous intoxication and lipid peroxidation in case of adenocarcinoma.

Phyllodes tumors of the breast: Analysis of 35 cases from a single institution

BackgroundPhyllodes tumors are rare fibroepithelial breast tumors with diverse biological behavior. Our study aim is to review the clinico-pathological features, prognostic factors and treatment outcome for patients presenting with phyllodes tumors of the breast to the Kuwait Cancer Control Center (KCCC). Patients and methodsWe retrospectively reviewed the clinical and pathological data of 35 women of histologically proved phyllodes tumors of the breast retrieved between January 1994 and December 2012. ResultsThe median age was 40years (21–63years). The median pathological tumor size was 6.8cm (3–25cm). Histologically, one patient (3%) presented with benign, 13 (37%) with borderline and 21 (60%) with malignant phyllodes. Twenty-eight patients (80%) were premenopausal. Twenty (57%) were ultimately treated with mastectomy (3 borderline, and 17 malignant) and 15 (43%) with conservative surgery (1 benign, 10 borderline and 4 malignant). Axillary staging was carried out in 9 patients (1 borderline and 8 malignant), none of them had nodal metastasis. Four patients with malignant phyllodes received postoperative radiotherapy. After a median follow-up period of 52months (range 5–211months), 5 developed local recurrence (1 benign, 2 borderline and 2 malignant). One patient with malignant phyllodes developed distant lung metastasis. The overall 5-year relapse free survival (RFS) was 74% (68% for borderline and 84% for malignant phyllodes). According to the treatment modality, the 5-year RFS was 69% for conservative surgery compared to 87% for mastectomy. It was 100% for irradiated patients versus 71% for non irradiated patients. ConclusionPhyllodes tumors are rare tumors that occur in relatively young women, when compared with the classical adenocarcinoma of the breast. They have a tendency to reach large sizes with absence of nodal metastasis. Although surgery is the mainstay of management, postoperative radiotherapy also appears to decrease the local recurrence rates in certain presentations.

Initial experience of pure 3-Dimensional laparoscopic cystectomy and urinary diversion

Objective To assess the perioperative safety and postoperative function of the pure 3-dimensional laparoscopic cystectomy and urinary diversion(P3DLC-UD). Methods From April 2014 to July 2015, P3DLC-UD was performed in 15 patients diagnosed with the bladder cancer in our center (orthotopic ileal neobladder for 8 cases and ileal conduit for 7 cases). Perioperative data, postoperative continence and overall survival rate were retrospectively analyzed. Results Fifteen patients underwent P3DLC-UD successfully as planned, with 8 patients underwent orthotopic ileal neobladder and 7 patients underwent ileal conduit. In neobladder group, the operative time were 300-600min (mean 428 min), estimated blood loss were 100-400ml (mean 210ml), and dissected lymph nodes were 11-29 (mean 16). One patient required blood transfusion (800ml) and one patient was diagnosed of constipation. The patients were followed up for a median period of 10 months (3-15 months). The renal function was normal with serum creatinine of 36.4-99.0 μmol/L (mean 77.3 μmol/L). One patient died of intestinal obstruction and 8 patients had no recurrence in neobladder group. As to postoperative continence, only 2 patients demanded 1 pad at daytime, while all patients needed 1 pad at nighttime. In ileal conduit group, the operative time were 300-390 min (mean 354 min), estimated blood loss were 50-400ml (mean 190ml), and dissected lymph nodes were 9-41 (mean 22), while 9-41 (mean 19) lymph nodes were got for all 15 cases. Two patients were diagnosed with urinary infection after the surgery. The patients were followed up for a median period of 5 months (1-9 months). The renal function was normal with serum creatinine of 36.4-74.0 μmol/L (mean 60.8 μmol/L). One patient died of cerebral infarction, and 1 patient found distant metastases in lung and died of cancer after chemotherapy during the follow-up period. There were no recurrent tumors of the other 5 patients in ileal conduit group. Conclusions P3DLC-UD is safe and feasible. More extensive, longer-term randomized trials are required to comprehensively assess the appropriateness and potential of this technique. Key words: Pure 3-dimensional laparoscopy; Bladder cancer; Orthotopic ileal neobladder; Ileal conduit

Prevention of Distant Lung Metastasis After Photodynamic Therapy Application in a Breast Cancer Tumor Model

The objective of this study was to investigate the activity of photodynamic therapy mediated by aluminum-chlorophthalocyanine contained in a polymeric nanostructured carrier composed by methyl vinyl ether-co-maleic anhydride (PVM/MA) against local subcutaneous breast cancer tumors and its effects against distant metastasis in a mouse tumor model. In our results, we observed a decrease in breast cancer tumor growth, prevention of distant lung metastases, and a significant increased survival in mice treated with photodynamic therapy. In addition to these results, we observed that tumor-bearing mice without treatment developed a significant extension of liver hematopoiesis that was significantly reduced in mice treated with photodynamic therapy. We hypothesized and showed that this reduction in (1) metastasis and (2) liver hematopoiesis may be related to the systemic activity of immature hematopoietic cells, specifically the myeloid-derived suppressor cells, which were suppressed in mice treated with photodynamic therapy. These cells produce a tolerogenic tumor environment that protects tumor tissues from immunological surveillance. Therefore, we suggest that photodynamic therapy could be employed in combination with other conventional therapies; such as surgery and radiotherapy, to improve the overall survival of patients diagnosed with breast cancer, as observed in our experimental resuIts.

14-3-3ζ promotes hepatocellular carcinoma venous metastasis by modulating hypoxia-inducible factor-1α.

Portal vein tumor thrombus (PVTT) is a type of intrahepatic metastasis arising from hepatocellular carcinoma (HCC) and is highly correlated with a poor prognosis. Hypoxia is common in solider tumors, including HCC, where it alters the behavior of HCC cells. We asked whether and how hypoxia contributes to PVTT formation. We demonstrated that increased intratumoral hypoxia is strongly associated with PVTT formation in HCC. We also showed that 14-3-3ζ is induced by hypoxia in HCC cells and correlates with PVTT formation in clinical HCC samples. In addition, 14-3-3ζ up-regulates HIF-1α expression by recruiting HDAC4, which prevents HIF-1α acetylation, thereby stabilizing the protein. Under hypoxic conditions in vitro, 14-3-3ζ knockdown inhibits hypoxia-induced HCC invasion by the HIF-1α/EMT pathway. Blockade of 14-3-3ζ in HCC cells reduces PVTT formation and distant lung metastasis in vivo. Moreover, a combination of 14-3-3ζ and HIF-1α expression is more prognostic for HCC patients than either protein alone. These results suggest that the hypoxia/14-3-3ζ/HIF-1α pathway plays an important role in PVTT formation and HCC metastasis.

Hepatitis C virus core protein interacts with Snail and histone deacetylases to promote the metastasis of hepatocellular carcinoma.

Downregulation of E-cadherin by the transcriptional repressor Snail is associated with acquisition of metastatic potential. Although hepatitis C virus (HCV) core protein has been implicated in hepatocarcinogenesis, it is unclear whether Snail is involved in HCV core-induced dysregulation of E-cadherin. Herein, we investigated the mechanism by which HCV core induces E-cadherin repression and the role of Snail in HCV core-mediated invasiveness and metastasis. We found that HCV infection, especially HCV core expression, effectively induced the epithelial-mesenchymal transition (EMT) in hepatoma cells by repressing E-cadherin. HCV core interacted with Snail and enhanced its binding to the E-box in the promoter region of E-cadherin, leading to decreased E-cadherin promoter activity. We found that HCV core, Snail, and the histone deacetylases HDAC1/HDAC2 formed a co-repressor complex at the E-cadherin promoter. Moreover, HCV core was shown to stabilize Snail through activation of the PI3K/Akt/GSK3β pathway. Silencing Snail expression restored E-cadherin expression and inhibited HCV core-promoted tumor growth and distant lung metastasis in vivo. Collectively, these results demonstrated that HCV core induced EMT by interacting with the transcriptional repressor complex Snail/HDACs at the E-cadherin promoter, which led to E-cadherin repression and increased invasiveness of hepatoma cells. These findings increase understanding of factors regulating metastasis in hepatoma and may ultimately lead to the development of novel treatment strategies for HCV-associated hepatocellular carcinoma.

Abstract 3958: The phenotypic and signaling consequences of a novel aberrantly spliced transcript of fibroblast growth factor receptor 3 in hepatocellular carcinoma

Abstract Aims: To identify the phenotypic and signaling consequences of a novel aberrantly spliced transcript of fibroblast growth factor receptor 3 (FGFR3) in hepatocellular carcinoma (HCC). Methods: The nested RT-PCR was employed to analyze the splicing of FGFR3 from 35 HCC cases and 11 cell lines. The bridging method was utilized to construct the sequence of FGFR3 Δ7-9 (deleting exon7, 8, and 9 from the full length of FGFR3 ORF). Based on the establishment of the FGFR3Δ7-9 lentiviral overexpress and knockdown models, the influence on bio-characters of HCC cell lines was inspected in vitro. The changes of tumorigenicity and metastases were examined quantitatively in vivo on nude mice using pathological study and Micro-PET/CT. Co-immunoprecipitation and surface plasmon resonance (SPR) binding analysis were employed to understand the ligand affinities between FGFR3Δ7-9 and FGFs. Interacting proteins and signal transduction pathway were detected by mass spectrometry and dual luciferase reporter assay. Results: FGFR3Δ7-9, lacking exons encoding the immunoglobulin-like domain III, was identified with high frequency in HCC. FGFR3Δ7-9 can apparently promote the proliferation, migration, and metastases of HCC cells both in vitro and in vivo. In contrast, knockdown of FGFR3Δ7-9 can restrict the abnormal malignancies of HCC cells. Co-immumoprecipation and SPR assay showed the binding affinity of FGFR3Δ7-9 to FGFs was significantly higher than that of wild type FGFR3 (P<0.01). More importantly, FGFR3Δ7-9 can be self-activated by self-dimerization and autophosphorylation even in the absence of ligand (FGFs). Consistent to the ligand-independence characteristic, FGFR3Δ7-9 demonstrated a stronger effect on inducing phosphorylation of ERK and AKT than wild type FGFR3, which may lead to abnormal down-stream PI3K/AKT/mTOR and ERK signal transductions. Furthermore, evidenced by tandem mass spectrum assay and Western blot, FGFR3Δ7-9 can up-regulate certain metastasis-related molecules: Snail and MMP-9. As a newly confirmed interaction protein to FGFR3Δ7-9, E-cadherin can be degraded via tyrosine phosphorylation, which may cause the abnormal cellular proliferation and migration consequently. Discussion/Conclusion: Dysregulation of mRNA splicing may generate an abnormal aberrant FGFR3 transcript, FGFR3Δ7-9. The joining of exon 6 to exon 10 in FGFR3Δ7-9 is in-frame, leading to the expression of a novel transmembrane form of FGFR3 containing an intact intracellular TK domain. As a ligand-independence or lowly ligand-dependent receptor, FGFR3Δ7-9 may function importantly in HCC tumorigenesis, proliferation, invasion and distant lung metastases. Altogether, our research strongly supports the idea that the dysregulation of FGFR3 expression by aberrant splicing of mRNA in a significant subset of HCC is an alternative pathway to neoplastic transformation. Note: This abstract was not presented at the meeting. Citation Format: Weihua Qiu, Weiping Yang, Xiaoqian Jing, Bingrui Wang, Xinyu Liu, Ding Ma, Helen Lin. The phenotypic and signaling consequences of a novel aberrantly spliced transcript of fibroblast growth factor receptor 3 in hepatocellular carcinoma. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3958. doi:10.1158/1538-7445.AM2015-3958

Hypoxia upregulates Rab11-family interacting protein 4 through HIF-1α to promote the metastasis of hepatocellular carcinoma.

Hypoxic microenvironment is a powerful driving force for the invasion and metastasis of hepatocellular carcinoma (HCC). Hypoxia-inducible factor 1α (HIF-1α), as a crucial regulator of transcriptional responses to hypoxia, induces the expression of multiple target genes involved in different steps of HCC metastatic process. It is critical to find target genes associated with metastasis under hypoxia for shedding new light on molecular mechanism of HCC metastasis. In this study, we uncovered that hypoxia could induce the upregulation of Rab11-family interacting protein 4 (Rab11-FIP4) and activation of Rab11-FIP4 promoter by HIF-1α. The overexpression of Rab11-FIP4 significantly enhanced the mobility and invasiveness of HCC cells in vitro, also contributed to distant lung metastasis in vivo, whereas silencing of Rab11-FIP4 decreased the ability of migration and invasion in HCC cells in vitro and suppressed lung metastasis in vivo. Rab11-FIP4 facilitated HCC metastasis through the phosphorylation of PRAS40, which was regulated by mTOR. Furthermore, the expression level of Rab11-FIP4 was significantly increased in HCC tissues and high expression of Rab11-FIP4 was closely correlated with vascular invasion and poor prognosis in HCC patients. A markedly positive correlation between the expression of Rab11-FIP4 and HIF-1α was observed in HCC tissues and combination of Rab11-FIP4 and HIF-1α was a more valuable predictor of poor prognosis for HCC patients. In conclusion, Rab11-FIP4 is a target gene of HIF-1α and has a pro-metastatic role in HCC, suggesting that Rab11-FIP4 may be a promising candidate target for HCC treatment.

Abstract LB-318: A direct in vivo RNAi screen unveils a myosin as potent tumor suppressor in head & neck squamous cell carcinomas

Abstract Modern genomics is revealing hundreds of alterations associated with cancer. Mining this information for cancer therapies is now predicated on weeding out ‘bystander’ alterations, identifying the ‘driver’ mutations responsible for tumor initiation and/or metastasis, and elucidating how these mutations alter the fundamental molecular pathways governing tissue growth. We, therefore, devise and employ a direct in vivo RNAi methodology in mice that allows us to simultaneously test hundreds of candidate human cancer genes. We focus on head and neck squamous cell carcinomas (HNSCCs), which arise from the epithelial lining of the upper aerodigestive tract, including the oral cavity, tongue, lip, pharynx and larynx. Despite their frequency, aggressive recurrence and poor prognosis (6th most common cancer; 50% mortality) HNSCCs are relatively poorly studied. Using non-invasive, ultrasound-guided in utero lentiviral-mediated delivery of RNAi, we selectively transduce the single-layered surface ectoderm of living E9.5 mouse embryos where stem cells stably incorporate and propagate the desired genetic alterations into adulthood in a mosaic fashion. To screen for putative driver mutations and to dissect the physiological relevance of epigenetic changes in gene expression that occur in the development of SCC tumor-initiating (stem) cells, we employed various tumor-susceptible mouse backgrounds and identified nine tumor suppressors, seven of which have not been hitherto linked to tumor development. Our top hit, Myh9, encodes the non-muscle myosin-IIa heavy chain (NMHCIIa). On tumor-susceptible backgrounds, epithelial-specific Myh9 RNAi or Myh9 knockout trigger formation of multiple invasive SCCs and even distant lung metastasis. Surprisingly, myosin-IIa’s function is manifested not only in conventional actin-related processes, but also in regulating DNA damage-induced, post-transcriptional p53 activation and p53 nuclear accumulation. Clinically, we show that ∼20% of human HNSCCs have lost myosin-IIa protein expression, ∼5% harbor evolutionarily conserved domain-specific MYH9 mutations, and that low MYH9 expression in HNSCCs correlates with poor survival (uni-variant Kaplan-Meier analysis). These findings establish novel SCC suppressors with potential prognostic and therapeutic relevance highlighting the utility of direct in vivo RNAi to integrate cancer genomics and mouse modeling to rapidly discover and validate potent but low penetrance cancer driver mutations. Importantly, our methodology tests function within the native environment and immune system prior to the gross perturbations in tissue architecture that invariably accompanies cancer progression. Citation Format: Daniel Schramek, Ataman Sondoel, Jeremy P. Segal, Evan Heller, Slobodan Beronja, Daniel Oristian, Boris Reva, Elaine Fuchs. A direct in vivo RNAi screen unveils a myosin as potent tumor suppressor in head & neck squamous cell carcinomas. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr LB-318. doi:10.1158/1538-7445.AM2014-LB-318

Usefulness of chest CT scan for head and neck cancer.

Chest computed tomography (CT) is not performed routinely or worldwide as the initial diagnostic procedure for patients with head and neck cancer (HNC). The significance of the chest CT scan for HNC diagnosis has not been thoroughly defined. The present study reports findings in an effort to broaden the acceptance and application of CT for HNC. Using medical records, we assessed initial chest CT scans of the patients with new-onset HNC between April 2004 and March 2010. The results were classified into three groups: nodules ≥ 1 cm, small nodules (< 1 cm) that were indeterminate and normal lungs without nodules. Lung nodules that increased in size and/or number at follow-up were regarded as malignant. First, the sensitivity of X-ray and CT for detection of lung nodules in patients with HNC was compared. Second, the nodules were estimated to be malignant or not malignant by follow-up chest CT. Third, statistical analyses were performed to determine the association between variables and distant lung metastases in patients with head and neck squamous cell carcinoma. In total, 332 patients underwent a chest CT scan as part of the initial examination. Lung nodules were detected on the initial chest CT in 77 patients: in contrast, lung nodules were detected on the initial chest X-ray in only five patients. On initial chest CT scans, lung nodules ≥ 1 cm were observed in 10 patients, small solitary lung nodules were observed in 67 patients, and lungs without nodules were observed in 255 patients. Lung nodules were detected in 77 (23.2%) patients, 25 (32.5%) of whom had malignant lung nodules. Moreover, in 18/67 patients (26.9%), small lung nodules initially classified as indeterminate were determined as malignant at follow-up. However, 30/255 patients (11.8%) without nodules at initial diagnosis developed lung nodules at follow-up. T3 or T4 disease (P = 0.006), N2 or N3 disease (P < 0.001) and stage 3 or 4 disease (P = 0.001) individually and significantly correlated with the development of lung metastases. In addition, lung nodules in initial CT scans (P = 0.004) and other distant metastases (P < 0.001) were significant predictors for the development of lung metastases at follow-up. Initial chest CT scan is recommended for patients with advanced HNC. Further, patients with advanced HNC with or without lung nodules, as detected on initial chest CT scans, should be followed up with additional scanning.