Cancer biochemistry biophysics最新文献

Native fluorescence emission and excitation spectra of SV40 infected human keratinocytes, A431 and SCC324 carcinoma cells, and normal human keratinocytes were measured and compared. A difference in the intracellular metabolic state of NADH was found between the normal cells and the cancer or virus-transformed cells. The observed difference, namely an increased proportion of bound, mitochondrial NADH in the cancer and virus-infected cells, manifests as a blue spectral shift in the emission spectra.

Echitamine chloride (EC), an indole alkaloid, extracted from the bark of Alstonia scholaris has got highly promising anticancer effect. The effect of this drug on the microsomal drug detoxifying system was studied in sarcoma-180 induced mice. When given sub-cutaneously at a dosage of 5 mg/kg body weight, it was able to alter the impaired drug detoxifying system which was observed in the Sarcoma-180 bearing mice. The levels of microsomal protein, Cyt-P450, Cyt-b5, NADH-Cyt-C-reductase, NADPH-Cyt-C-reductase, and glu-6 phosphatase were determined. The levels of these drug metabolizing enzymes were decreased in S-180 bearing mice. EC treatment corrected to near normal levels of these enzymes and microsomal hemeproteins. In order to understand the mechanism responsible for the decreased protein level and its normalization after treatment with EC, 3H-Phenylalanine incorporation study was carried out. From the results, it is observed that the synthesis of apoproteins is also altered in tumor-bearing animals. All these changes which were observed in tumor-bearing animals were corrected to near normal levels after treatment with EC.

Free-radical-mediated damages may play an important role during metastasis. To investigate their relevance in the metastatic process MDA levels, glutathione peroxidase (GPX) and superoxide dismutase (SOD) activities, and selenium, zinc and copper contents were determined in plasma and erythrocytes from 20 cancerous patients with metastasis and 30 age-matched controls. Significantly higher concentrations of MDA in plasma as well as in erythrocytes were found comparing to the control group. In both plasma and erythrocytes, GPX activity and selenium and zinc levels were significantly lower in patients than in controls. However, SOD activity in erythrocytes and copper levels in both plasma and erythrocytes were significantly higher in patients. The impaired antioxidant system may favor accumulation of free radicals which may induce the process of metastasis. On the other hand, it is possible that the antioxidant system is impaired as a consequence of abnormality in the antioxidative metabolisms due to the cancer process.

Cell-matrix interactions have important effects on phenotypic features, such as morphology, differentiation and cell growth. Several papers have suggested that when cell-matrix interactions are interrupted, cells grow as multicellular spheroids and eventually undergo apoptosis. We found that when ET(-), a laminin non-adherent colon cancer cell line, was cultured on poly-2-hydroxyethyl methacrylate (HEMA) coated plastic, the cells floated as cellular aggregates of spheroids or as single cells. Some of the single cells contained a very large intracytoplasmic lumen (ICL) and appeared similar to signet ring cells. These ICL were lined by a layer of short microvilli. The number of the cell did not increased cells when cultured on poly-HEMA. Another type of single cells, usually without ICL, demonstrated the characteristics of apoptotic cells by histologic examination. Acridine orange staining, flow cytometry and electron microscopy confirmed the apoptotic nature of those cells. In immunohistochemical staining for proliferating cell nuclear antigen, spheroids of cells and single cells with ICL were immunoreactive, while most of the single cells without ICL were negative. These results suggest that multicellular aggregation and formation of ICL were induced by the adaptation of ET(-) colon cancer cells in a harmful environment caused by reduced adhesiveness, and these changes might be related to cell survival.

Carnitine has two main functions, i.e., transporting long-chain fatty acids into the mitochondrial matrix for beta-oxidation to provide cellular energy and modulating the rise in intramitochondrial acyl-CoA/CoA ratio, which relieves the inhibition of many intramitochondrial enzymes involving glucose and amino acid catabolism. The present study examined the acid soluble carnitine (ASCAR) acid insoluble carnitine (AICAR) and total carnitine (TCAR) concentrations of 50 human brain tumor tissues and 11 normal brain tissues. The ASCAR levels significantly higher in gliomas and meningiomas than brain, however similar to brain in metastatic adenocarcinomas. AICAR levels were lower than brain in all tumors with the exception of a medullablastoma. TCAR levels were similar to brain in all tumor types. Decreased AICAR levels may be due to increased utilization of lipids or enhanced phospholipid and cholesterol synthesis which is need for increased membrane synthesis or formation of eicosanoids. Also decreased concentrations may be a reflection of camitine and its acylesters role in preserving the physiologic membrane structure function from oxidative damage.

Combined antitumor activity of CPT-11 and 5-fluorouracil (5-FU) was evaluated in a human cultured cell line derived from lung cancer. After 24 h culture with SN-38 followed by 5-FU 24 h, synergistic effect was observed in the cell line. In addition, the antitumor effect of this combination was studied in in vivo experiments using Donryu rat with Yoshida sarcoma cells. CPT-11 and 5-FU synergistically inhibited tumor growth. There was no significant increase of toxicity as assessed by the body weights. These results might support for the combination with 5-FU and CPT-11 in a chemotherapy for cancer.

Reverse micelles were employed to test the accuracy of the widely accepted mechanism for alpha-chymotrypsin in a highly structured aqueous system similar to intracellular conditions. Results yielded from spectrophotometrical assays of the alpha-chymotrypsin catalyzed hydrolysis of both p-nitrophenyl acetate (p-NPA) and p-nitrophenyl trimethylacetate (p-NPTA) were kinetically analyzed to determine constants typical of the proposed mechanistic model. This was accomplished through the establishment of a control, i.e. the well studied buffer system, for comparison between the reverse micellular environment and a bulk aqueous solution. Control group results yielded kinetic constants in favor of the proposed mechanism (Km = 1.55 x 10(-5) +/- 1.40 x 10(-6) M for p-NPA and a Km = 4.97 x 10(-6) +/- 2.29 x 10(-7) M, Km(app) = 4.92 x 10(-6) +/- 2.33 x 10(-8) M, k2 = 4.34 x 10(-3) +/- 1.31 x 10(-3), k(cat) = 1.96 x 10(-3) +/- 2.47 x 10(-4), and Ks = 1.60 x 10(-5) +/- 4.61 x 10(-6) M for p-NPTA). In contrast, similar reactions of the enzyme in a reverse micellular system produced kinetic constants atypical to that representative of the textbook mechanism. (Km = 1.59 x 10(-4) +/- 2.70 x 10(-5) M, Ks = -8.67 x 10(-5) +/- 4.46 x 10(-5) M and Km(app) = -4.80 x 10(-5) +/- 7.05 x 10(-5) M for p-NPA and Km = 1.95 x 10(-4) +/- 9.28 x 10(-5) M, Km(app) = -1.79 x 10(-4) +/- 2.36 x 10(-5) M, and Ks = -3.95 x 10(-4) +/- 1.18 x 10(-4) M for p-NPTA). In addition to negative kinetic constants, alpha-chymotrypsin seemed to display characteristics indicative of super-activity and a hysteretic response. Overall, the widely accepted mechanism for alpha-chymotrypsin appeared to fail within the confines of reverse micelles, due to the direct influence of the system's highly structured form.

Several studies demonstrated that certain fatty acids have specific effects on tumor cells. n-3 series fatty acids (alpha-linolenic acid, eicosapentaenoic acid and docosahexaenoic acid) may suppress the carcinogenesis, whereas n-6 series fatty acids (arachidonic acid, linoleic acid) may exert tumor promoting effects. In this study, 19 patients with various brain tumors and 12 control brain tissues were studied. n-3, n-6, n-9 unsaturated fatty acids and certain saturated fatty acids levels were measured in the plasma membrane of tumor or control brain tissues by capillary gas chromatography. We found that the level of docosahexaenoic acid from n-3 series fatty acids was significantly lower in gliomas and meningiomas than controls (p = 0.000). Total n-3 fatty acids level was also significantly lower in tumors than controls (p = 0.000). The levels of linoleic acid, arachidonic acid and dihomogamma linolenic acid from n-6 series were significantly higher in gliomas and meningiomas compared with controls (p = 0.000). Total n-6 fatty acids level was also significantly higher in tumors than controls (p = 0.000). Furthermore, in total n-9 fatty acids, total unsaturated fatty acids and total saturated fatty acids levels, there were no significant differences in gliomas and meningiomas compared with controls (p = 0.6840, p = 0.4388 and p = 0.4343, respectively). This findings suggest that n-6 fatty acids can act as a tumor-promoting agent in human brain tumors.

We have previously established a vincristine resistant human lung cancer cell line (PC-9/VCR) by a stepwise exposure of parental line PC-9 to vincristine. In this study the resistant cells showed enhanced vincristine cytotoxicity in the presence of cytochalasin B and D. The increase in cytotoxicity was associated with an enhanced accumulation and a reduced efflux of vincristine. Colchicine and taxol had no effects on vincristine accumulation. Several cytoplasmic proteins were overexpressed in the resistant cells. The two major ones, with molecular weights of 58.8 kDa and 83.2 kDa, were shown by western blotting to be beta-tubulin and actin, respectively. The polymerized tubulin level in the resistant cells was significantly (p < 0.05) higher than that in the parental cells. These results suggest that the cellular cytoskeletons might play an important role in VCR resistance in the PC-9/VCR human lung cancer cell line.

In this study, breast cancer (n = 23) and benign breast disease (n = 15) patients were evaluated in relation to oxidative stress. The extent of lipid peroxidation was assessed by measuring thiobarbituric acid reactive substances (TBARS) in plasma. Erythrocyte glutathione peroxidase (GSH Px), CuZn speroxide dismutase (CuZn SOD), glutathione (GSH) and plasma vitamin E, cholesterol Fe, Zn, Cu levels were analysed in both groups GSH Px (p < 0.01), vitamin E (p < 0.001), Zn (p < 0.01), Cu (p < 0.05) and cholesterol (p < 0.01) concentrations were found to be significantly increased, TBARS level (p < 0.01) significantly decreased in breast cancer patients in comparison to benign breast disease group.