Application of in vitro soft agar techniques for growth of tumor cells to the study of colon cancer

Buick, Roger; Fry, Stephen E.; Salmon, Sydney E.

doi:10.1002/1097-0142(19800315)45:5+<1238::aid-cncr2820451333>3.0.co;2-r

Cited by 30 publications

(6 citation statements)

References 9 publications

(1 reference statement)

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“…Several investigators have utilized the labeling index as a method for determining the efficacy of anticancer agents [ 14,151. Another test utilized incorporation of ['Hluridine [16]. The most popular assay in recent times has been the clonogenic assay, which is used to determine the effect of anticancer agents on the sur-vival of human tumor cells grown in soft agar [17][18][19][20], a medium that inhibits the growth of fibroblasts and permits cultivation of clonogenic cells. The effect of anticancer agents on DNA synthesis has recently been evaluated .…”

Section: Introductionmentioning

confidence: 51%

Cyclophosphamide activity against B16 melanoma in a rapid in vitro test system

Hill

Hill²

1983

Journal of Surgical Oncology

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An in vitro test system is described for evaluation of the response of tumor cells to chemotherapeutic agents. The test is simple and rapid and does not require the use of sterile technique. The response of B16 melanoma in vivo in C57BL/6 mice to single graded doses of cyclophosphamide is compared to the response of B16 tumor cells in vitro to cyclophosphamide that has been activated in vivo and recovered in plasma. The assay involves preincubating the tumor cells with the activated drug and subsequently measuring the ability of the cells to incorporate [3H]dThd ([3H] thymidine) into acid-insoluble material. In vivo activated drug is maximally effective in 20 min and the entire test can be completed within 8 hr. This test may be useful as a rapid preliminary screening method to predict the effect of chemotherapy on cancer.

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Section: Introductionmentioning

confidence: 51%

Cyclophosphamide activity against B16 melanoma in a rapid in vitro test system

Hill

Hill²

1983

Journal of Surgical Oncology

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“…Colony-forming efficiency in soft agar was determined as described previously (Buick et al, 1980) with minor modifications. NIH/3T3 cells transfected with HCCR-2 (5 Â 10 3 cells) were suspended in a 1 ml mixture containing 0.3% noble agar (Difco Laboratories Inc., Detroit, MI, USA), complete medium and 10% FBS, and layered on a 2 ml solidified base of 0.6% noble agar in complete medium with 10% FBS in 35 mm plates.…”

Section: Differential Displaymentioning

confidence: 97%

Identification and differential expression of novel human cervical cancer oncogene HCCR-2 in human cancers and its involvement in p53 stabilization

et al. 2003

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Basic studies of oncogenesis have demonstrated that either the elevated production of particular oncogene proteins or the occurrence of qualitative abnormalities in oncogenes can contribute to neoplastic cellular transformation. The purpose of this study was to identify unique oncogenes that are differentially expressed in human cancers and characterize their functions in tumorigenesis. To discover new putative oncogenes, the differential display RT-PCR method was applied using normal cervical tissues, cervical cancer cell lines, cervical cancer tissues, and metastatic tissues. We identified a new human cervical cancer oncogene HCCR-2 that was overexpressed in various human tumors including leukemia, lymphoma, and carcinomas of the breast, kidney, ovary, stomach, colon, and uterine cervix. Ectopic expression of HCCR-2 resulted in direct tumorigenic conversions of NIH/3T3 and Rat1 fibroblasts. Nude mice injected with NIH/3T3 cells stably transfected with HCCR-2 formed tumors in 4 weeks. The resultant tumors display characteristics of an epithelial carcinoma. In HCCR-2 transfected NCI-H460 cells and RKO cells, stabilization of the p53 tumor suppressor occurred without genetic mutation and correlated with functional impairment, as indicated by the defective induction of p53-induced p21 WAF1 , MDM2, and bax. These results indicate that HCCR-2 probably represents a new oncogene that is related to tumorigenesis, functioning as a negative regulator of the p53 tumor suppressor.

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“…15 Agar assays can also distinguish tumor cells from nontransformed cells, because cells lacking the ability to undergo anchorage-independent growth are unable to thrive on an agar substrate. 16,17 Previous work in our laboratory demonstrated that the growth of nonmetastatic tumor cells on agar could be restricted when the concentration of the agar medium was increased from 0.3% (soft agar) to 0.6% (hard agar).…”

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confidence: 99%

Selection of Brain Metastasis-Initiating Breast Cancer Cells Determined by Growth on Hard Agar

Guo

Fan

Zhang

et al. 2011

The American Journal of Pathology

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An approach that facilitates rapid isolation and characterization of tumor cells with enhanced metastatic po-The vast majority of cancer deaths result from progressive growth of metastases that are resistant to conventional therapies. 1 Metastases originate from a selected subpopulation of cells that reside in a biologically heterogeneous primary tumor. 2,3 Results from experimental 4 and clinical 5 examinations indicate that most metastases are clonal in origin and that the metastatic process is highly selective. 6 Studies have also shown profound differences between local and disseminated cancers, 7 suggesting that information on primary tumors alone may not be sufficient to determine optimal therapeutic interventions. For this reason, researchers have directed considerable effort toward improving understanding of the molecular phenotypes of metastasis-initiating tumor cells.One widely used experimental approach to isolate populations of tumor cells with enhanced metastatic potential is an in vivo selection process in which tumor cells are implanted into syngeneic or immunodeficient mice and metastasis is allowed to occur. Tumor cells from the resultant metastatic lesions can be isolated and expanded to establish cell sublines, some of which may have higher metastatic capacity than the parental tumorcell population. 8 Comparative gene expression profiling of parental tumor cells and their metastatic subpopulations has yielded invaluable information regarding the genetic determinants critical for organ-specific metastasis. 9 For example, Kang et al 10 compared the transcriptional profiles of parental MDA-231 human breast cancer cells with those of a bone-colonizing variant of this cell line and reported the underlying gene expression signature required for organ tropism to bone. Investigators used a similar approach to identify genes whose expression is critical for metastasis of breast cancer cells to the brain 11 and lungs. 12 Nevertheless, although this in vivo selection technique has provided new insight into the cellular and molecular mechanisms that control site-specific metastasis, there are some disadvantages. Perhaps the principal drawback of in vivo selection is that it can be time-consuming,

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Application of in vitro soft agar techniques for growth of tumor cells to the study of colon cancer

Cited by 30 publications

References 9 publications

Cyclophosphamide activity against B16 melanoma in a rapid in vitro test system

Cyclophosphamide activity against B16 melanoma in a rapid in vitro test system

Identification and differential expression of novel human cervical cancer oncogene HCCR-2 in human cancers and its involvement in p53 stabilization

Selection of Brain Metastasis-Initiating Breast Cancer Cells Determined by Growth on Hard Agar

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