Telomere length is a novel predictive biomarker of sensitivity to anti-EGFR therapy in metastatic colorectal cancer.

BACKGROUND: Telomeres are TTAGGG tandem repeats capping chromosomal ends and partially controlled by the telomerase enzyme. The EGFR pathway putatively regulates telomerase function, prompting an investigation of telomere length (TL) and its association with anti-epidermal growth factor receptor (EGFR) therapy in metastatic colorectal cancer (mCRC). METHODS: Colorectal cancer cell lines were treated with multiple drugs and sensitivity determined. Clinical information was gathered from 75 patients who had received anti-EGFR drugs. Telomere length was measured using a validated qRT-PCR technique. RESULTS: In CRC cell lines, TL independently predicted cetuximab sensitivity. Cells with shorter TL had growth inhibition of 18.6±3.41% as compared with 41.39±8.58% in longer TL (P=0.02). These in vitro findings were validated clinically, in a robust multivariate model. Among patients with KRas WT tumours, those with longer TL had a superior median progression-free survival (PFS) of 24.9 weeks than those with shorter TL; median 11.1 weeks, HR 0.31; P=0.048. CONCLUSION: Telomere length could be a potential unique biomarker predictive of clinical benefit (PFS) of mCRC patients treated with anti-EGFR therapy. This is the novel demonstration of a complex hitherto undescribed interaction, placing anti-EGFR therapy, EGFR pathway, and the telomerase complex within a clinical context.

Silymarin inhibited proliferation and induced apoptosis in hepatic cancer cells.

OBJECTIVES: The aim of this study was to investigate mechanisms involved in the growth inhibitory effect of silymarin, in humanhepatocellular carcinoma. MATERIALS AND METHODS: The human hepatocellular carcinoma cell line HepG2 was utilized and the MTT assay was performed to study the antiproliferative effect of silymarin. Dual staining was undertaken for ethidium bromide/acridine orange, propidium iodide staining and DNA fragmentation studies were executed to confirm the presence of apoptosis. Cell-cycle analysis was revealed by flow cytometry and mitochondrial transmembrane potential was measured by uptake of the mitochondrial-specific lipophilic cationic dye rhodamine 123. Western blotting analysis for cytochrome c, p53, Bax, Bcl-2, APAF-1, caspase-3, survivin, beta-catenin, cyclin D1, c-Myc and PCNA was carried out. RESULTS: Silymarin inhibited population growth of the hepatocellular carcinoma cells in a dose-dependent manner, and the percentage of apoptotic cells was increased after treatment with 50 and 75 microg/ml silymarin for 24 h. Silymarin treatment increased the proportion of cells with reduced DNA content (sub-G(0)/G(1) or A(0) peak), indicative of apoptosis with loss of cells in the G(1) phase. Silymarin also decreased mitochondrial transmembrane potential of the cells, thereby increasing levels of cytosolic cytochrome c while up-regulating expression of pro-apoptotic proteins (such as p53, Bax, APAF-1 and caspase-3) with concomitant decrease in anti-apoptotic proteins (Bcl-2 and survivin) and proliferation-associated proteins (beta-catenin, cyclin D1, c-Myc and PCNA). CONCLUSIONS: Our results demonstrate that silymarin treatment inhibited proliferation and induced apoptosis in the human hepatocellular carcinoma cell line HepG2.

Effect of silymarin on N-nitrosodiethylamine induced hepatocarcinogenesis in rats.

AIM: To study the effect of silymarin on the levels of tumor markers and MDA (malondialdehyde)-DNA adduct formation during N-nitrosodiethylamine induced hepatocellular carcinoma in male Wistar albino rats. METHODS: The levels of AFP, CEA and activities of liver marker enzymes in serum, MDA-DNA immunohistochemistry were done according to standard procedures in the control and experimental groups of rats. RESULTS: Hepatocellular carcinoma was evidenced from significant (p < 0.05) increases of alpha-fetoprotein, carcinoembryonic antigen, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, acid phosphatase, lactate dehydrogenase, gamma-glutamyltransferase and 5'-nucleotidase in serum and increased MDA-DNA adducts were also observed in the tissue sections of hepatocellular carcinoma. Silymarin treatment significantly attenuated the alteration of these markers and decreased the levels of MDA-DNA adduct formation. CONCLUSION: Silymarin could be developed as a promising chemotherapeutic adjuvant for the treatment of liver cancer.

Protein factor which induces conversion between Physarum ornithine decarboxylase forms in vitro.

The rapid activity modulation of ornithine decarboxylase (L-ornithine carboxy-lyase, EC 4.1.1.17) in Physarum polycephalum is closely correlated with a reversible post-translational modification of this enzyme. A factor has now been isolated from homogenates of exponentially-grown microplasmodia which catalyzes the conversion of the active enzyme form, A, into its less active, B state. Partial purification of this A-B converting factor has been achieved using DEAE-Sephacel chromatography and Ultrogel AcA-34 gel filtration. It appears to be a heat labile, acidic protein with a molecular weight of about 35 000 which binds to large macromolecules in crude fractions isolated using low ionic strength buffers. The in vitro converting reaction requires the presence of spermidine or spermine (1 mM) while putrescine is much less effective, and inorganic cations are ineffective at levels up to 5 mM. Enzyme conversion is reduced in elevated ionic strengths and in the presence of polyamine chelators such as ATP, ADP or GTP (1.0 mM). Under current assay conditions the interaction between ornithine decarboxylase and this factor is stoichiometric, yet it is not reversed even by conditions which favor dissociation of protein-protein interactions. This is the first report of the isolation of a protein factor which is involved in the interconversion of ornithine decarboxylase between its alternate enzyme states.