Establishment Of Xuanwei Lung Adenocarcinoma Cancer Cell Line XLA-0610 And The Preliminary Assay Of The Side Population Cell(SP) In The Lung Cancer Line

BANKGROUND&OBJECTIVE:Lung cancer is currently one of the most common malignant tumor all over the world.In China,the morbidity and mortality of the disease has also shown an upward trend.Moreover,there is a high incidence of lung cancer in Xuanwei,Yunnan province,where the mortality rate of this disease in the female is the highest.To establish the female lung adenocarcinoma cell line in Xuanwei region of Yunnan Province,and provide an ideal experimental model for the study of preventing and curing lung cancer.And to conduct a preliminary analysis of fthe SP cells for this strain of cells and further to research for the relevant characteristic of the cancer stem cell.It provides new opportunities and molecular targets for the diagnosis and treatment of the female lung adenocarcinoma cell line in Xuanwei region of Yunnan ProvinceMETHODS:1.The cell line derived from a surgical specimen of lung cancer.After primary culture,its biologic characteristics were studied with both light and electron microscope,determination of doubling time and growth curve,culturing in soft agar, flow cytometry(FCM),chromosome and G-band as well as C-12 multi-tumor marker detection.2.Preparing for cell suspension from which is being established lung adenocarcinoma cell line of XLA-0610,staining with Hoechst33342 dye and antagonists verapamil,and conducting a preliminary assay of the SP cells for this strain of cells by flow cytometry. RESULT:1.Morphological study,proliferation dynamics and immersed growth demonstrated the cultured cells' malignant characteristics,doubling time of cell is h and cloning efficiency is 36.3%.Chromosome and its G-band analysis revealed a chromosome number ranging from 61to 72,with a mode number of 64 to 69.C-12 multi-tumor markers were detected:Lysis buffer of cultured cells:CA125 390.12↑, CA15-3 86.35↑;Supernatant of cultured cells:CA125 43.08↑.The cell line wasn't contaminated by mycoplasma.2.The result of a preliminary assay of the SP cells in XLA—0610 by flow cytometry:The percentage of SP cells cell line is about 14.9%.After Verapamil blocked,the percentage of SP cells obviously decreases and are about 2.0%CONCLUSIONS:1.The cell line is according to the rules of establishing a cell line in vitro,The study contents and detection means of the biological characteristics of the cell line are more comprehensive.XLA-0610 is proved to be a newly-built human lung adenocarcinoma cell line.Thus it can provide a good experimental model for further studie of lung cancer.2.Draw a conduction by the preliminary assay of the SP cells:XLA-0610 cell line exists SP cells and the percentage of SP cells is about 14.9%.After Verapamil blocked,the percentage of SP cells obviously decreases and are about 2.0%.It has significant difference from the non-blocking group by verapamil.It prompted the existence of lung cancer stem cells.That the preliminary assay of the SP cells provides some scientific reference value for diagnosis of lung cancer,the identification and purification of CSC.