Growth Potential Of Tumor Cells Research Articles

Identifying spatial immuno-oncological parameters of mouse tumors using Imaging Mass Cytometry

Abstract Identification of immuno-oncological (IO) processes that dictate tumor growth, metastasis, and immune response is essential for selecting promising drug candidates in preclinical models. Imaging Mass Cytometry™ (IMC™) is a proven high-plex imaging technology that enables deep characterization of the complexity of tumor tissue without disrupting spatial context while eliminating artifacts caused by tissue autofluorescence. Here, we showcase the Maxpar® OnDemand™ Mouse Immuno-Oncology IMC Panel Kit for application on mouse tumor tissues. We compiled a 33-plex antibody panel to quantitatively assess IO-related processes and applied it to a tissue microarray containing a large variety of mouse tumors. We digitized high-plex data from mouse tissues using the Hyperion™ Imaging System and generated images demonstrating the detailed layout of the tumor microenvironment (TME). We further conducted single-cell analysis to identify specific populations of tumor and immune cells in the TME. The Mouse IO IMC Panel Kit successfully identified pathophysiological processes such as immune cell infiltration and activation, signaling pathway activation, biomarkers of epithelial-to-mesenchymal transition (EMT), metabolic activity, growth, and the TME tissue architecture. Single-cell analysis of several highly relevant tumor types separated distinct cellular clusters representing tumor, immune, stromal, and vascular cells. Our quantitative analysis of tumor composition revealed critical insights regarding prognostic parameters such as metastatic and growth potential of tumor cells and activity of immune cell infiltrates. For Research Use Only. Not for use in diagnostic procedures.

Astroprincin (FAM171A1, C10orf38): A Regulator of Human Cell Shape and Invasive Growth

Our group originally found and cloned cDNA for a 98-kDa type 1 transmembrane glycoprotein of unknown function. Because of its abundant expression in astrocytes, it was called the protein astroprincin (APCN). Two thirds of the evolutionarily conserved protein is intracytoplasmic, whereas the extracellular domain carries two N-glycosidic side chains. APCN is physiologically expressed in placental trophoblasts, skeletal and hearth muscle, and kidney and pancreas. Overexpression of APCN (cDNA) in various cell lines induced sprouting of slender projections, whereas knockdown of APCN expression by siRNA caused disappearance of actin stress fibers. Immunohistochemical staining of human cancers for endogenous APCN showed elevated expression in invasive tumor cells compared with intratumoral cells. Human melanoma cells (SK-MEL-28) transfected with APCN cDNA acquired the ability of invasive growth in semisolid medium (Matrigel) not seen with control cells. A conserved carboxyterminal stretch of 21amino acids was found to be essential for APCN to induce cell sprouting and invasive growth. Yeast two-hybrid screening revealed several interactive partners, of which ornithine decarboxylase antizyme-1, NEEP21 (NSG1), and ADAM10 were validated by coimmunoprecipitation. This is the first functional description of APCN. These data show that APCN regulates the dynamics of the actin cytoskeletal and, thereby, the cell shape and invasive growth potential of tumor cells.

Genetic dissection of colorectal cancer progression by orthotopic transplantation of engineered cancer organoids

In the adenoma-carcinoma sequence, it is proposed that intestinal polyps evolve through a set of defined mutations toward metastatic colorectal cancer (CRC). Here, we dissect this adenoma-carcinoma sequence in vivo by using an orthotopic organoid transplantation model of human colon organoids engineered to harbor different CRC mutation combinations. We demonstrate that sequential accumulation of oncogenic mutations in Wnt, EGFR, P53, and TGF-β signaling pathways facilitates efficient tumor growth, migration, and metastatic colonization. We show that reconstitution of specific niche signals can restore metastatic growth potential of tumor cells lacking one of the oncogenic mutations. Our findings imply that the ability to metastasize-i.e., to colonize distant sites-is the direct consequence of the loss of dependency on specific niche signals.

Editorial: Gamma Knife radiosurgery and nonfunctioning pituitary adenomas

The publication of Lee et al. is a useful and well-written addition to the literature.1 They analyze, based on a review of previous studies and their own experience, the advantages and disadvantages of Gamma Knife radiosurgery (GKRS) in the treatment of patients with clinically nonfunctioning pituitary adenomas. The results of GKRS are also compared with those produced by conventional resection. Clinically nonfunctioning pituitary adenomas represent the most frequent pituitary adenoma type among pituitary tumors. Medical treatment is not available, and pituitary tumor irradiation has several significant disadvantages. Thus, the question is, Gamma Knife surgery or conventional resection? The authors conclude that in older patients (older than 70 years) and in patients with other comorbidities in whom surgical intervention is contraindicated, initial GKRS is an acceptable alternative. Obviously, as the authors emphasize, an experienced team is needed to decide which treatment to use and to undertake the GKRS. One great disadvantage of GKRS is that the tumor is not investigated by pathologists. Morphological investigation is of crucial importance. Several diseases can mimic pituitary adenomas: for example, other primary intrasellar neoplasms, metastatic tumors, inflammatory diseases, and others. We have seen several metastatic carcinomas arising from the lung, colon, and other organs, which were mis diagnosed as pituitary adenomas! Another reason morphological investigation is important is that it allows one to draw conclusions regarding the growth potential of tumor cells. We can assess the Ki 67 nuclear labeling index, a valuable method of revealing the cell proliferation rate. Maybe in the future new drugs will be developed, which will permit medical therapy. Advances in molecular/genetic methods may provide an alternative treatment option as well. Obviously, more studies are needed to decide whether one should use GKRS and in which patients. (http://thejns.org/doi/abs/10.3171/2013.10.JNS132248) Disclosure

Activation of antioxidant pathways in ras-mediated oncogenic transformation of human surface ovarian epithelial cells revealed by functional proteomics and mass spectrometry.

Cellular transformation is a complex process involving genetic alterations associated with multiple signaling pathways. Development of a transformation model using defined genetic elements has provided an opportunity to elucidate the role of oncogenes and tumor suppressor genes in the initiation and development of ovarian cancer. To study the cellular and molecular mechanisms of Ras-mediated oncogenic transformation of ovarian epithelial cells, we used a proteomic approach involving two-dimensional electrophoresis and mass spectrometry to profile two ovarian epithelial cell lines, one immortalized with SV40 T/t antigens and the human catalytic subunit of telomerase and the other transformed with an additional oncogenic ras(V12) allele. Of approximately 2200 observed protein spots, we have identified >30 protein targets that showed significant changes between the immortalized and transformed cell lines using peptide mass fingerprinting. Among these identified targets, one most notable group of proteins altered significantly consists of enzymes involved in cellular redox balance. Detailed analysis of these protein targets suggests that activation of Ras-signaling pathways increases the threshold of reactive oxidative species (ROS) tolerance by up-regulating the overall antioxidant capacity of cells, especially in mitochondria. This enhanced antioxidant capacity protects the transformed cells from high levels of ROS associated with the uncontrolled growth potential of tumor cells. It is conceivable that an enhanced antioxidation capability may constitute a common mechanism for tumor cells to evade apoptosis induced by oxidative stresses at high ROS levels.

PROLIFERATING ACTIVITY OF GASTRIC-CARCINOMA INVADING THE MUSCULARIS PROPRIA AND METASTASIS TO LYMPH-NODES

From 1965 to 1987, 126 patients who had gastric cancer invading the muscularis propria (pm) were treated with curative resection in the Department of Surgery II, Kyushu University. We examined the correlation between the proliferating activity of the tumor by staining the tissue for proliferating cell nuclear antigen (PCNA) and the occurrence of lymph node metastasis present in 46.8% (59/126) of the patients. Undifferentiated carcinoma was more frequent in these patients and lymphatic and vascular involvement was prominent. Patients with a lymph node metastasis had a higher level of PCNA staining in the tumors than did those without lymph node metastasis (labeling index: 36.5 +/- 9.6% vs. 25.3 +/- 8.3%) (P < 0.01). The survival rate for patients with lymph node metastasis was lower than that for those without this metastasis (P < 0.01). The 10-year survival rate was 63.6% for the patients with lymph node metastasis and 86.8% for those without lymph node metastasis. Thus, the presence of lymph node metastasis is associated with a high growth potential of tumor cells in pm gastric cancer. Close follow-up and adjuvant chemotherapy in the postoperative period is recommended for these patients.

Metastatic potential of hyperplastic alveolar nodule derived mouse mammary tumor cells following intravenous inoculation*

A comparison was made of the in vivo growth potential of tumor cells derived from mouse mammary adenocarcinomas arising in premalignant outgrowth lines D 1 and D 2 . Twenty-one tumors were dissociated and cultured as cell monolayers in vitro . Three to four days later 50,000–100,000 cells were inoculated intravenously into syngeneic female BALBlc mice. Primary cultures of tumor cells which arose in D 2 HAN outgrowths varied considerably in their ability to form tumor nodules in the lungs of syngeneic mice 4 weeks post-inoculation. An increase in either the number of cells inoculated or in the length of in vivo incubation influenced the extent of tumor growth observed in the lungs. Primary cultures of tumor cells which arose in D 1 HAN outgrowths inoculated at the same cell number rarely formed tumor nodules in the lungs 4 weeks post-injection. If, however, twice as many cells were introduced, a small number of large nodules were observed within 4 weeks. An epithelial cell line (WAZ- 2 T) isolated from D 1 tumor tissue produced small lung nodules by 4 weeks, but larger more numerous nodules arose when in vivo incubation was extended an additional 3 weeks. This study indicates that primary D 2 tumor cells have a variable but higher degree of metastatic potential than primary D 1 tumor cells following intravenous inoculation. Tumors which develop in the D 2 and D 1 premalignant lines may be useful for studies of chemotherapy and cell latency of tumor metastases and the identification of the specific cell subpopulations involved in these events.

Metastatic potential of hyperplastic alveolar nodule derived mouse mammary tumor cells following intravenous inoculation

A comparison was made of the in vivo growth potential of tumor cells derived from mouse mammary adenocarcinomas arising in premalignant outgrowth lines D 1 and D 2. Twenty-one tumors were dissociated and cultured as cell monolayers in vitro. Three to four days later 50,000–100,000 cells were inoculated intravenously into syngeneic female BALBlc mice. Primary cultures of tumor cells which arose in D 2 HAN outgrowths varied considerably in their ability to form tumor nodules in the lungs of syngeneic mice 4 weeks post-inoculation. An increase in either the number of cells inoculated or in the length of in vivo incubation influenced the extent of tumor growth observed in the lungs. Primary cultures of tumor cells which arose in D 1 HAN outgrowths inoculated at the same cell number rarely formed tumor nodules in the lungs 4 weeks post-injection. If, however, twice as many cells were introduced, a small number of large nodules were observed within 4 weeks. An epithelial cell line (WAZ- 2T) isolated from D 1 tumor tissue produced small lung nodules by 4 weeks, but larger more numerous nodules arose when in vivo incubation was extended an additional 3 weeks. This study indicates that primary D 2 tumor cells have a variable but higher degree of metastatic potential than primary D 1 tumor cells following intravenous inoculation. Tumors which develop in the D 2 and D 1 premalignant lines may be useful for studies of chemotherapy and cell latency of tumor metastases and the identification of the specific cell subpopulations involved in these events.

Effect of radiation on canine intestinal anastomoses.

RADIATION therapy as a preoperative adjuvant in the surgical treatment of patients with a variety of malignant neoplasms, including carcinoma of the colon and rectum demands continued interest and evaluation. The basic purpose of such combined therapy is to improve the cure rate and survival time for patients so treated beyond that attained by excisional therapy alone. Possible advantages of a course of preoperative radiation therapy include (1) a decrease in the size of the tumor mass, which may include both an inflammatory as well as a neoplastic component, thus increasing the resectability rate, and, as a corollary, fostering a less traumatic removal of the cancer bearing organ, and (2) a reduction in the viability of neoplastic cells thus diminishing the implantation ability and growth potential of tumor cells inoculated locally or disseminated intravascularly at the time of operation. There is evidence, both experimental and clinical, to support the