Tumors colonies

Shoemaker, R.H. et al., Application of a human tumor colony-forming assay to new drug screening, Cancer Res., 45, 2145-2153, 1985. 

Moved] Cranberry fruit of Early Black cultivar was fractionated chromatographically and fractions were analyzed for flavonoid content. The effects of the flavonoid fractions and ursolic acid, an abundant triterpenoid in cranberry peel, were assessed in two models of colon cancer and one model of breast cancer. Clonogenic soft agar assays were used to determine the effect of these compounds on tumor colony formation in HCT-116, HT-29 and MCF-7 cells. MTT and trypan blue assays were performed to assess their ability to inhibit tumor cell proliferation. TUNEL assays were performed to assess apop-totic response to the cranberry compounds. The proanthocyanidins inhibited tumor colony formation in HCT-116 and HT-29 cells in a dose-dependent manner, with greater effect on the HCT-116 cell line. Ursolic acid strongly inhibited tumor colony formation in both colon cell lines. These compounds also decreased proliferation in all three tumor cell lines with the HCT-116 cell line most strongly affected. (150 words)... 

Tumor colonies that differ in color from the lung parenchyma (melanoma) can be counted with the aid of a dissecting microscope. 

Most pulmonary tumor colonies in mice are in the pleura. Therefore, pigmented colonies (such as melanomas) can be counted without lung dissection. However, it may be more facile, especially when there are extensive pulmonary metastases, to dissect the lungs into the individual four or five lobes which are then enumerated individually using a dissecting microscope. 

Tumor colonies that are not pigmented and are minimally different in their color from the pulmonary parenchyma are more difficult to enumerate. The following method is one of several that can be used to induce contrast between tumor colonies and lung parenchyma. 

Animals suspected of having tumor colonies in an organ are killed, and their lungs or other organs removed and rinsed in tap water. 

The lung parenchyma will turn yellow and 24 h later, the white tumor colonies are readily distinguished from the yellow lung parenchyma. 

The lungs are rinsed in water to remove excess Bouin s solution, and the tumor colonies counted with the aid of a dissecting microscope (see Note 9). Murine lungs with greater than 300 colonies should be reported as >300 or too numerous to count. 

No rodent bioassay In vitro Neumega did not stimulate growth of tumor colony-forming cells harvested from patients with a variety of malignancies... 

An in vitro. system that is a good predictor of human clinical activity is the human-tumor-colony-forming a.ssay (HTCFA). This system uses fre.sh human tumor tissue from individual patients. It is valuable in selecting chemotherapeutic agents for individual tumor types and occasionally specific patients, but its use in large-scale primary screening has not been fea.sible. 

LLC-bearing mice had tumor metastasis to the liver, with about 50 tumors colonies per mouse Fig. (6) . Tuna triacylglycerols (1000 or 2000 mg/kg) significantly reduced the number of tumor cell colonies that metastasized to the liver compared with the number in intrasplenic untreated LLC-bearing mice. The growth of metastatic tumors in the livers of LLC-bearing mice was also inhibited by orally administered tuna triacylglycerols (1000 or 2000 mg/kg) Fig. (6) . 

The mice with subcutaneously implanted LLC cells showed tumor metastasis to the lungs. EPAD (1000 mg/kg) reduced the number of metastatic tumor colonies in the lungs compared with that in the untreated LLC-bearing mice, but EPA ethylester (300 or 1000 mg/kg) and EPAD (300 mg/kg) had no effect Table (4) . 

Izquierdo MA, Degen D, Myers L, et al. Effects of the hematopoietic growth factors GM-CSF, IL-3, and IL-6 on human tumor colony-forming units taken directly from patients. Ann Oncol 1995 6 927-32. 

Hanauske U, Hanauske A-R, Marshall MH, et al. 1987. Biphasic effect of vanadium salts on in vitro tumor colony growth. Int J Cell Cloning 5 170-178. 

Table 7. Effects of oleic acid on tbe spleen weight and number of metastatic tumor colonies in the livers of mice with intrasplenically implanted LLC1...

Treatment No of animals Spleen weight (mg) Metastatic tumor colonies in the liver... 

Treatment n Tumor weight (mg) Metastasis to the lung (number of (number of mice tumor colonies) with metastasis) ... 

Experimental pulmonary metastasis was assessed by tumor cell injection into the lateral tail vein of mice. C57BL/6 mice were injected intravenously (IV) with B16-BL6 melanoma (5 X 10 ) admixed with or without chitin derivatives. The mice were killed 14 days after tumor inoculation, and the number of lung tumor colonies was counted. In a spontaneous pulmonary metastasis assay, mice were inoculated subcutaneously with B16-BL6 cells (5 X 10 ) in the right hind footpad. The primary tumors were surgically removed on day 21 after tumor inoculation. The injection of chitin derivatives was performed on various days before or after the amputation. Mice were killed 14 days after the amputation, and the number of lung tumor colonies was counted. An outline of the experiment is shown in Scheme 3. 

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