Pine needle oil induces G2/M arrest of HepG2 cells by activating the ATM pathway.

Over the last two decades, inducing DNA damage of cancer cells by natural medicines has become a research hotspot in the field of cancer treatment. Although various natural medicines have anticancer effects, very few studies have been conducted to explore the anti-cancer effect of pine needle oil. In the present study, the role of pine needle oil in inducing G2/M arrest in HepG2 cells was investigated. The data revealed that pine needle oil could induce DNA damage in a dose-dependent manner. In the pine needle oil-treated HepG2 cells, the protein levels of phosphorylated (p)-ataxia-telangiectasia mutated (ATM), γ-H2A histone family, member X, p-p53, p-checkpoint kinase 2 and p-cell division cycle 25C were evidently increased, indicating that pine needle oil facilitated G2/M arrest in HepG2 cells through the ATM pathway. In response to the treatment with pine needle oil, ATM was activated in HepG2 cells, which subsequently phosphorylated downstream targets and induced G2/M arrest. In summary, the data of the present study indicated that pine needle oil induces G2/M arrest in HepG2 cells by facilitating ATM activation.

Measurement of hydroxyproline in collagen with three different methods.

Determination of the 4-hydroxy-l-proline (hydroxyproline) concentration may provide useful information for the diagnosis and prognosis of diseases caused by disorders of collagen metabolism. The objective of the present study was to apply liquid chromatography-mass spectrometry (LC-MS) to assess the hydroxyproline concentration. The hydroxyproline concentration in lung and liver tissues measured by LC-MS was compared with values obtained by a colorimetric method, as well as a fluorescence method using high-performance liquid chromatography (HPLC) from previous studies by our group. The determination of the hydroxyproline concentration by LC-MS was improved as compared with that using the colorimetric and HPLC methods, due to its simplicity, high sensitivity (pg level) and short separation time. These results suggested that utilizing the LC-MS method for measuring the hydroxyproline concentration would be advantageous for the diagnosis of diseases associated with abnormalities of collagen metabolism.