To clarify the mechanism of the anticarcinogenic effect of conjugated linoleic acid (CLA), its intracellular distribution needs to be determined. Subcellular fractionation using centrifugation techniques is a method that is frequently used for isolation of cell organelles from different tissues. But as the size and density of the organelles differ, the method needs to be optimised for every type of tissue. The novelty of this study is the application of a subcellular fractionation method to human healthy and cancerous renal and testicular tissue. Separation of total tissue homogenate into nuclei, cytosol, and a mixture of mitochondria and plasma membranes was achieved by differential centrifugation. As mitochondria and plasma membranes seemed to be too similar in size and weight to be separated by differential centrifugation, discontinuous density-gradient centrifugation was carried out successfully. The purity of the subcellular fractions was checked by measuring the activity of marker enzymes. All fractions were highly enriched in their corresponding marker enzyme. However, the nuclear fractions of kidney and renal cell carcinoma were slightly contaminated with mitochondria and plasma membrane fractions of all tissues with lysosomes. The fraction designated the cytosolic fraction contained not only cytosol, but also microsomes and lysosomes. The CLA contents of the subcellular fractions were in the range 0.13-0.37% of total fatty acids and were lowest in the plasma membrane fractions of all types of tissue studied. C16:0, C18:0, C18:1 c9, C18:2 n-6, and C20:4 n-6 were found to be the major fatty acids in all the subcellular fractions studied. However, marked variations in fatty acid content between subcellular fractions and between types of tissue were detectable. Because of these differences between tissues, no general statement on characteristic fatty acid profiles of single subcellular fractions is possible.

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On fine needle histological sampling tissue fragments are obtained, making possible a histological diagnosis with a more precise tumour description. The purpose of our study was to examine the differential diagnostic value of ultrasonically guided fine needle aspiration combined with histological

In this study the FA compositions of healthy and cancerous human renal tissues from the same patients are compared with special reference to the CLA and PUFA content. CLA was preferentially incorporated into neutral lipid compared with phospholipid classes. Its distribution profile was similar to that of monounsaturated FA, but unlike that found with 18:2n-6. Different incorporation patterns were found for individual CLA isomers. Comparing renal cell carcinoma (RCC) and healthy kidney, the total CLA content was significantly lower in the cholesterylester fraction and significantly higher in the PE and PS fractions from RCC. The most significant differences between healthy and cancerous renal tissue were in the content of t10,c12-CLA. Furthermore, the lipid class distributions of n-6 PUFA were determined, and several significant differences between RCC and healthy renal tissue were found. This is of interest, as it has been proposed that the anticarcinogenic properties of dietary CLA are associated with their interference in the metabolism of 20:4n-6. The involvement of CLA in preventing renal cancer cannot be definitively demonstrated from the design of this study, nor was it intended, but the complete determination of the FA composition of adjacent healthy and cancerous tissues may provide an insight if lipids are involved in this disease.

Differences in the FA composition of subcellular fractions from healthy and cancerous kidney tissues from the same patients were examined. Only minor differences in CLA content were found between the healthy and the cancerous tissue portions. Regarding the distribution pattern, CLA incorporation into nuclei and cytosol was significantly higher than incorporation into plasma membranes and mitochondria, which could be correlated to the neutral lipid content of these fractions. The subcellular distribution pattern of CLA was similar to that observed with monounsaturated FA but unlike that found with 18:2n-6, which underlines the different physiological properties of CLA and 18:2n-6. Because PUFA have been suggested to have an effect on cancer risk, the contents of n-3 and n-6 PUFA were determined in kidney and renal cell carcinoma (RCC). The 18:2n-6 content and delta5 desaturase activity were significantly lower, and the 18:3n-6, 20:3n-6, and 20:5n-3 contents and delta6 desaturase activity were significantly higher in RCC than in healthy renal tissue, indicating a changed PUFA metabolism in RCC. Previous research has suggested that CLA inhibits the elongation and desaturation of 18:2n-6 into 20:4n-6. In that case, one might speculate that a diet enriched in CLA would be a useful tool in preventing RCC. However, the involvement of CLA in preventing renal cancer could not be demonstrated definitively from the design of this experiment. Further understanding of the cause and/or consequence of the difference in FA metabolism may lead to a better understanding of RCC.