Differentiation-based histologic grading of colorectal carcinoma (CRC) is widely used, but its clinical impact is limited by insufficient prognostic value, interobserver disagreement, and the difficulty of its application to CRC with specific histologic types such as mucinous and medullary carcinoma. A recently proposed novel grading system based on quantifying poorly differentiated clusters (PDCs) claims to have the advantages of reproducibility and improved prognostic value, and might apply to heterogeneous CRC. We aimed to validate the clinicopathologic significance of the PDCs-based grading system and to determine the relationship between this grading system and microsatellite instability (MSI). Two hundred and one patients who had undergone radical surgery were reviewed. Based on the number of PDCs, 85, 58, and 58 tumors were classified as grade (G) 1 (42.3%), G2 (28.9%), and G3 (28.9%), respectively. PDCs-based grade was significantly associated with T, N, and M stages; lymphovascular invasion; conventional histologic grade; and frequent tumor budding (all P <0.001). In multivariate analysis, PDCs-based grade was found to be an independent prognostic factor for disease-free survival (P = 0.022; hazard ratio, 3.709 [G2], 7.461 [G3]). G3 CRC significantly correlated with high MSI (MSI-H) compared to G1 and G2 (P = 0.002; odds ratio, 5.750). In conclusion, this novel grading would provide valuable prognostic information to a greater number of patients and would require continued verification. PDCs-based grading is feasible for CRCs with heterogeneous morphology, and we propose that the association between G3 and MSI-H be further evaluated in different histological subtypes of CRC.
Colorectal Neoplasms/*genetics/*pathology/surgery ; Disease-Free Survival ; Female ; Humans ; Lymphatic Metastasis/pathology ; Male ; *Microsatellite Instability ; Middle Aged ; Neoplasm Grading/*methods ; Tumor Burden/*physiology

BACKGROUNDThe presence of residual tumor after surgery for pituitary adenoma may necessitate further treatment. The suprasellar and parasellar extension of the tumor have been widely considered as the predictors for residual tumor. However there is scarcity of studies regarding the preoperative tumor volume and residual tumor. This study was conducted to evaluate if tumor volume could predict the outcome of transsphenoidal pituitary surgery.

METHODSA prospective study was designed and 48 patients who underwent transsphenoidal pituitary surgery within 1 year in the First Affiliated Hospital of Xi'an Jiaotong University were included in this study. The preoperative tumor volume and immediate postoperative tumor volume (within 4 - 7 days) were calculated in the contrast magnetic resonance imaging by using the formula of ellipsoid. All these volumes were divided into three subgroups, i.e. group 1, group 2 and group 3 with preoperative volume of less than 4 cm(3), 4 - 8 cm(3), and more than 8 cm(3) respectively. The parasellar and suprasellar extension of the tumor were also classified by Knosp and modified Hardy's classifications.

RESULTSBaseline characteristics were comparable. The preoperative tumor volume of more than 8 cm(3) (group 3, (12.1 ± 1.1) cm(3)) had increased risk on postoperative tumor residue (P < 0.01) than the other two groups ((2.1 ± 0.3) cm(3) and (6.1 ± 0.3) cm(3) in groups 1 and 2). The mean postoperative volume in group 3 patients ((2.2 ± 0.1) cm(3)) was significantly higher than the other two groups (P < 0.01).

CONCLUSIONPreoperative volume of more than 8 cm(3) can be considered as a predictor for postoperative residual volume.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pituitary Gland ; pathology ; surgery ; Pituitary Neoplasms ; pathology ; surgery ; Prospective Studies ; Treatment Outcome ; Tumor Burden ; physiology ; Young Adult

OBJECTIVETo generate an oncolytic herpes simplex virus (oHSV) permissive mouse melanoma cell line B16RHSV, preserving the tumorigenic ability in syngeneic mice.

METHODSThe herpes simplex virus entry mediator (HVEM) gene was amplified by PCR from human melanoma cell line A375, and cloned into pGEM-T Easy vector for sequencing. The HVEM gene was then cloned into pcDNA3 vector to generate pcDNA3-HVEM for transfection of mouse melanoma cell line B16-F10 cells. After that, the putative transfected cells were selected in full growth medium containing G418. The HVEM-expressing cells were isolated by immunomagnetic bead separation. The mouse melanoma cell line expressing oHSV receptor-HVEM, designated as B16RHSV, was generated. The permissibility of B16RHSV cells to oHSV infection was examined with green fluorescence protein (GFP)-expressing oHSV (oHSVGFP). To investigate the tumorigenic ability of both cells in vivo, 2×10(5) cells in 100 µl were subcutaneously inoculated into the right flanks of C57/BL mice.

RESULTSIn vitro, the B16RHSV mouse melanoma cells were shown by fluorescence microscopy capable of being infected by oHSVGFP. In vivo, the B16RHSV cells, like their wild type counterpart, grew to form melanoma in syngeneic mice.

CONCLUSIONA herpes simplex virus-permissive mouse melanoma cell line was established. Its tumorigenicity remained unchanged.

Animals ; Cell Line, Tumor ; Female ; Gene Amplification ; Genetic Vectors ; Herpesvirus 1, Human ; genetics ; physiology ; Humans ; Melanoma ; pathology ; virology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Plasmids ; Receptors, Tumor Necrosis Factor, Member 14 ; genetics ; metabolism ; Transfection ; Tumor Burden

OBJECTIVETo study the anti-tumor effects of Newcastle disease virus (NDV) strain D817 on human colon carcinoma model in nude mice.

METHODSThe nude mouse model of human colon carcinoma was established by subcutaneous inoculation of human colon cancer LOVO cells. The tumor-bearing mice were given PBS, 5-Fu, high-dose NDV D817, moderate-dose NDV D817 or low-dose NDV D817 via caudal vein injection. The tumor size and weight of mice were measured. The liver damages were examined by histopathology. Apoptosis and necrosis of tumor cells were detected by flow cytometry. The endotumoral content of TNF-alpha was detected using a mouse TNF-alpha ELISA kit. The live virus was detected by hemagglutination (HA) test.

RESULTSThe moderate-dose NDV D817 inhibited the tumor growth more apparently than 5-Fu. The tumor growth inhibition rate reached to 48.1%. The liver damage and the weight change caused by NDV were less severe. NDV D817 made an increased apoptosis index and induced production of TNF-alpha. Live virus was not detected in important organs except in the tumor of nude mice by HA test.

CONCLUSIONIn the anti-tumor process in nude mice bearing xenografts of human colon carcinoma, a suitable dose of NDV D817 is more safe and effective.

Animals ; Apoptosis ; Cell Line, Tumor ; Colonic Neoplasms ; pathology ; therapy ; Humans ; Liver ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Newcastle disease virus ; physiology ; Random Allocation ; Tumor Burden ; Tumor Necrosis Factor-alpha ; metabolism ; Xenograft Model Antitumor Assays

The bones are the most common sites of breast cancer metastasis. Upon arrival within the bone microenvironment, breast cancer cells coordinate the activities of stromal cells, resulting in an increase in osteoclast activity and bone matrix degradation. In late stages of bone metastasis, breast cancer cells induce apoptosis in osteoblasts, which further exacerbates bone loss. However, in early stages, breast cancer cells induce osteoblasts to secrete inflammatory cytokines purported to drive tumor progression. To more thoroughly evaluate the role of osteoblasts in early stages of breast cancer metastasis to the bones, we used green fluorescent protein-labeled human breast cancer cell lines MDA-MB-231 and MDA-MB-435, which both induce osteolysis after intra-femoral injection in athymic mice, and the murine pre-osteoblastic cell line MC3T3-E1 to modulate osteoblast populations at the sites of breast cancer metastasis. Breast cancer cells were injected directly into the femur with or without equal numbers of MC3T3-E1 cells. Tumors grew significantly larger when co-injected with breast cancer cells and MC3T3-E1 cells than injected with breast cancer cells alone. Osteolysis was induced in both groups, indicating that MC3T3-E1 cells did not block the ability of breast cancer cells to cause bone destruction. MC3T3-E1 cells promoted tumor growth out of the bone into the extraosseous stroma. These data suggest that breast cancer cells and osteoblasts communicate during early stages of bone metastasis and promote tumor growth.
Animals ; Bone Neoplasms ; secondary ; Breast Neoplasms ; pathology ; Cell Line ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Femur ; pathology ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Osteoblasts ; cytology ; physiology ; Osteolysis ; etiology ; Tumor Burden

OBJECTIVETo investigate the effect of LIM and SH3 protein 1 (LASP-1) expression on the proliferative ability of human colorectal cancer cells in vitro and in vivo.

METHODSRT-PCR and Western blot were used to screen cells of the colorectal cancer cell line with no or with minimal endogenous LASP-1 expression. LASP-1 cDNA with or without GFP was transfected into SW480 colorectal cancer cells with minimal LASP-1 expression. Stable transfectants were established after G418 selection. Cell proliferative capacity was assessed by MTT assay. Tumor growth was visualized by whole-body imaging system.

RESULTSpcDNA3-LASP-1 and pEGFP-LASP-1 vectors were successfully constructed and transfected into the SW480 cells. After comparative study for 7 days, LASP-1 over-expression was found capable of enhancing significantly the proliferation of colorectal cancer cells in vitro. Stable transfectants with GFP expression were inoculated subcutaneously into the nude mice. Tumorigenesis and proliferation ability of LASP-1-overexpressed transfectants were higher than those of the control cells.

CONCLUSIONLASP-1 gene expression enhances proliferation of colorectal cancer cells and may serve as a useful marker for colorectal cancer progression.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; physiology ; Animals ; Biomarkers, Tumor ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; Cytoskeletal Proteins ; genetics ; metabolism ; physiology ; DNA, Complementary ; genetics ; Gene Expression Regulation, Neoplastic ; Green Fluorescent Proteins ; metabolism ; Humans ; LIM Domain Proteins ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Plasmids ; RNA, Messenger ; metabolism ; Recombinant Proteins ; metabolism ; Tumor Burden

OBJECTIVETo study the effect of CEA gene regulation on the anti-tumor activity of oncolytic adenovirus H101 to esophageal carcinoma, and to explore the intrinsic factors influencing H101 sensitivity.

METHODSStable human esophageal cancer cell line EC9706 cells with lower (EC9706-SCEA) and higher CEA expression (EC9706-CEA) were chosen, thawed and cultured, and then to analyse the influence of CEA expressed at different levels on cell growth. The cytotoxic effect of H101 was assayed by in vitro and nude mouse in vivo.

RESULTSThe cell growth experiment showed that the population doubling time of EC9706-SCEA, EC9706-CEA and EC9706 cells were (30.9 ± 2.0) h, (31.1 ± 2.5) h and (29.1 ± 2.6) h, respectively, showing no significant difference among them (P > 0.05). The cytotoxic activity of H101 was higher on EC9706-SCEA than on other four groups, when MOI was ≥ 0.01 PFU (P < 0.05). The mouse experiment showed that H101 inhibited the growth of transplanted tumors in all experimental groups. Its effect on CEA-silenced tumors (inhibition rate was 61.5% to 74.5%) was significantly higher than that on CEA-overexpression tumors (32.3% to 38.5%) and control EC9706 transplanted tumors (35.5% to 44.8%). There was a significant difference between them (P < 0.05).

CONCLUSIONSThe results in vitro and in vivo experiments show that H101 can enhance the cytotoxic effect on EC9706 cells with lower CEA expression. To silence the expression of CEA may provide a novel strategy for target gene therapy of esophageal carcinoma.

Adenoviridae ; physiology ; Animals ; Carcinoembryonic Antigen ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Esophageal Neoplasms ; metabolism ; pathology ; therapy ; Female ; Gene Silencing ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Oncolytic Virotherapy ; Oncolytic Viruses ; physiology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Tumor Burden

OBJECTIVEThe aim of this study was to construct a new oncolytic virus oHSV2hGM-CSF and evaluate its oncolytic activity in vitro and in vivo in parallel with oHSV1hGM-CSF.

METHODSoHSV2hGM-CSF was a replication-competent, attenuated HSV2 based on the HG52 virus (an HSV2 strain). It was engineered to be specific for cancer by deletion of the viral genes ICP34.5 and ICP47 and insertion of the gene encoding hGM-CSF. To measure the in vitro killing effect of the virus, 15 human tumor cell lines (HeLa, Eca-109, PG, HepG2, SK/FU, CNE-2Z, PC-3, SK-OV3, A-549, 786-0, MCF-7, Hep-2, HT-29, SK-Mel-28, U87-MG) and mouse melanoma (B16R) cell line were seeded into 24-well plates and infected with viruses at MOI = 1 (multiplicity of infection, MOI), or left uninfected. The cells were harvested 24 and 48 hours post infection, and observed under the microscope. For animal studies, the oncolytic viruses were administered intratumorally (at 3-day interval) at a dose of 2.3 x 10(6) PFU (plaque forming unit, PFU) for three times when the tumor volume reached 7-8 mm3. The tumor volume was measured at 3-day intervals and animal survival was recorded.

RESULTSBoth oHSV2hCM-CSFand oHSV1hGM-CSF induced widespread cytopathic effects at 24 h after infection. OHSV2hGM-CSF, by contrast, produced more plaques with a syncytial phenotype than oHSV1hGM-CSF. In the in vitro killing experiments for the cell lines HeLa, HepG2, SK-Mel-28, B16R and U87-MG, oHSV2hGM-CSF eradicated significantly more cells than oHSV1hGM-CSF under the same conditions. For the mouse experiments, it was observed that oHSV2hGM-CSF significantly inhibited the tumor growth. At 15 days after B16R tumor cells inoculation, the tumor volumes of the PBS, oHSV1hGCM-CSF and oHSV2hGM-CSF groups were (374.7 +/- 128.24) mm3, (128.23 +/- 45.32) mm3 (P < 0.05, vs. PBS group) or (10.06 +/- 5.1) mm3 (P < 0.01, vs. PBS group), respectively (mean +/- error). The long term therapeutic effect of oHSV2hGM-CSF on the B16R animal model was evaluated by recording animal survival over 110 days after tumor cells inoculation whereas all the mice in the PBS group died by day 22 (P < 0.01). The anti-tumor mechanism of the newly constructed oHSV2hGM-CSF against B16R cell tumor appeared to include the directly oncolytic activity and the induction of anti-tumor immunity to some degree.

CONCLUSIONThe findings of our study demonstrate that the newly constructed oHSV2hGM-CSF has potent anti-tumor activity in vitro to many tumor cell lines and in vive to the transplanted B16R tumor models.

Animals ; Cell Line, Tumor ; Female ; Gene Deletion ; Genetic Engineering ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Herpesvirus 2, Human ; genetics ; immunology ; Humans ; Immediate-Early Proteins ; genetics ; metabolism ; Melanoma, Experimental ; pathology ; therapy ; virology ; Mice ; Mice, Inbred C57BL ; Oncolytic Virotherapy ; methods ; Oncolytic Viruses ; genetics ; physiology ; Random Allocation ; Tumor Burden ; Viral Proteins ; genetics ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo confirm the role of Tiam1 (T lymphoma invasion and metastasis 1) gene in the proliferation and metastasis of colorectal cancer.

METHODSProliferative and metastatic abilities of Tiam1 transfectant were investigated by subcutaneous injection of cells and surgical orthotopic transplantation (SOI) in mice.

RESULTSThe expression of Tiam1 led to a pronounced increase in HT29/Tiam1 cell growth starting from day 7, up to 2.5 fold increase of tumor volume at day 20 post injection. Tumors in the HT29/Tiam1 group receiving surgical orthotopic implantation were significantly heavier than those in HT29/mock group (t = -14.916, P < 0.01). In vivo metastasis assay by SOI showed that in HT29/Tiam1 group, 7/7 of mice developed peritoneal metastases and 4/7 had hepatic lesions. In addition, one of the seven HT29/Tiam1 group mice had tumors in lung, spleen and lymph nodes. In the HT29/mock group, only 2/7 of animals had peritoneal metastases and none produced detectable tumor in the liver.

CONCLUSIONSTiam1 gene plays an important role in the proliferation, invasion and metastasis of colorectal cancer. It may serve as a useful clinical marker for tumor progression and metastasis of colorectal cancer.

Animals ; Biomarkers, Tumor ; analysis ; Cell Proliferation ; Colorectal Neoplasms ; pathology ; Guanine Nucleotide Exchange Factors ; genetics ; metabolism ; physiology ; HT29 Cells ; Humans ; Liver Neoplasms ; secondary ; Lung Neoplasms ; secondary ; Lymphatic Metastasis ; Mice ; Mice, Nude ; Neoplasm Invasiveness ; Neoplasm Transplantation ; Peritoneal Neoplasms ; secondary ; Plasmids ; T-Lymphoma Invasion and Metastasis-inducing Protein 1 ; Transfection ; Tumor Burden

OBJECTIVETo observe the influence of hypoxia-inducible factor 1 alpha (HIF-1alpha) on angiogenesis in prostate carcinoma in vivo and to investigate its molecular mechanism.

METHODSLNCaP/HIF-1alpha and LNCaP cells were cultured, the level of PSA in the supernatant of the culture medium detected by ELISA assay before and after the transfection, and the cellular cycle measured by flow cytometry. Nude mouse models of subcutaneous tumor were established with LNCaP/HIF-1alpha and LNCaP cells, the tumor growth observed, and tumor specimens collected for immunohistochemical staining.

RESULTSCompared with the LNCaP cells, LNCaP/HIF-1alpha cells showed an obviously decreased PSA level (t = 8.243, P < 0.05) and enhanced proliferous activity. The tumorigenesis rate increased and the tumorigenesis time advanced in the LNCaP/HIF-1alpha group of the nude mice. Immunohistochemistry displayed higher expressions of VEGF, iNOS and Ang-2 in the LNCaP/HIF-1alpha than in the LNCaP group.

CONCLUSIONThe over-expression of HIF-1alpha can up-regulate VEGF and iNOS involved in angiogenesis in vivo and contribute to the invasive potency of LNCaP cells. HIF-1alpha may have no influence on Ang-2 either in vitro or in vivo, while the expression of Ang-2 is regulated by other factors in vivo.

Animals ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; physiology ; Immunohistochemistry ; Male ; Mice ; Mice, Nude ; Neoplasms, Experimental ; blood supply ; metabolism ; pathology ; Neovascularization, Pathologic ; physiopathology ; Nitric Oxide Synthase Type II ; biosynthesis ; Prostatic Neoplasms ; blood supply ; genetics ; pathology ; Transfection ; Transplantation, Heterologous ; Tumor Burden ; Vascular Endothelial Growth Factor A ; biosynthesis