The Study Of Primary Cell Incubation Of Lung Cancer And Its Breath Test

Objectives: To explore the methods of incubating the primary lung cancer cells and to detect volatile organic compounds (VOCs) in the primary lung cancer cells as tumor markers, as well as to provide evidences for breath test of lung cancer. Methods: We took twenty lung specimens from operations done from June, 2008 to December,2008 and took the tumor tissue as the experimental group, while the periphery tissue more than ten centimetes far from the tumor was taken as the control. Incubated them until above half of the bottom covered with cells and then test the VOCs levels. Samples from the headspace of the culture vessel were collected with simultaneous preconcentration by adsorption on solid sorbents and subsequently thermodesorbed for analysis by gas chromatography mass spectrometry (GC-MS).Results: There are totally 20 cases, including 9 cases of squamous carcinoma, 7 cases of adenocaricinoma, 2 cases of adeno-squamous carcinoma and 2 cases of benign lesions. It is available to invert the culture flask for 30 minutes before routine cultivation, which is benefit for adherence. After the tissue is cut, it is immediately inoculated. Twenty-four to forty-eight hours later, there are cells visible around the brink of the tissue. The average culture time for twenty cases is 11.75 days, and for adenocarcinoma is 10.1 days (71days/ 7cases), which is less than that for squamous carcinoma 14.1 days (127 days/9 cases) (P<0.05). Seen from the mass-spectrogram, it can be found that the content of the substances whose appearance time are respectively 17.78 minutes and 18.75 minutes are obviously less in the blank medium control than those in the culture fluid for the primary lung cancer cells. Moreover, the contents of these substances are different in different types of lung cancer, which can be seen from the fact that the contents in adenocarcinoma, bronchiolar carcinoma and squamous carcinoma are increasing in order; the content of the substance whose appearance time is 18.75 minutes is much more in squamous carcinoma. Similarity search showed that the peak time of 17.78 minutes and 18.75 minutes are 1 - methyl - naphthalene and pentadecane respectively. Conclusions: The methods for culturing the primary lung cancer cells is available and spare. This research implied that 1-methyl-naphthalene and pentadecane, which are produced through the metabolization of the lung cancer cells, may be the useful biomarkers to classify the lung cancer types.