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Screening of Lung Cancer Stem Cell Surface Markers and Researching Enrichment Methods of Lung Cancer Stem Cell

Author: YueWenKe
Tutor: ZhouQingHua
School: Tianjin Medical University
Course: Surgery
Keywords: Lung cancer stem cells Flow cytometry Serum-free culture Induced by cisplatin CD133 CD24
CLC: R734.2
Type: Master's thesis
Year: 2011
Downloads: 161
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Abstract

Objective: This study analyzed by flow cytometry of CD133, CD24, CD44 and CD221 expression in different types of human lung cancer cell lines, screening of lung cancer stem cell surface markers, serum-free suspension culture into a ball and cisplatin induced enriched in lung cancer cell lines of lung cancer stem cells and detection screening to the surface signs molecules in which the expression in order to access to be able to effectively separated lung cancer stem cells of the surface signs molecules lay a experimental basis for the subsequent human lung tumor stem cells in the screening and functional studies. Methods: flow cytometry CD133, CD24, CD44 and CD221 in human lung adenocarcinoma cell line A549 squamous carcinoma cell line YTMLC-9, with the same genetic background in human lung large cell lung cancer with high metastatic potential of cell lines L9981 and low transfer potential cell strains NL9980 expression screening lung cancer stem cell marker; study by the serum-free suspension culture and induced by cisplatin in L9981 L9981 and A549 lung cancer cells obtained sphere. L9981 cells and serum-free culture into a ball of L9981 cells cultured in serum-containing adherent compare both biological morphology under the microscope, the application of the Vi-cell cell viability analyzer counting cells and draw growth curves compare both proliferation ability by Transwell experimental study their invasive ability differences in the in vivo tumorigenicity in nude mice observed both to study the biology of lung cancer spheroids verify its dry. The same method to identify the biological function of the A549. CD133 and CD24 expression levels in the lung cancer stem cell-like cell subsets and further to detect lung cancer stem cells as a possible separation markers feasibility to verify the CD133 and CD24. Results: 1. The streaming single parameter test results display higher levels of CD221 and CD44 expression in A549, YTMLC-9, L9981 and NL9980 lung cancer cell lines, CD133 and CD24 expression levels in various lung cancer cell lines. Flow dual-parameter test results showed that of CD133-CD24-CD133-CD24 of CD24-CD221, CD221 CD24 in various cell lines express higher levels of CD133 CD24 of CD133 of CD44, CD221-CD24, CD221 of CD24 expression levels is low. CD221 expression in A549 and YTMLC-9 cells, the levels were not significantly different, with different metastatic potential in the large cell lung cancer cell line L9981 and NL9980 expression in two significant differences (P lt; 0.01). 2.A549 cells in serum-free suspension culture balling poor L9981 into the ball significantly. L9981 cells in suspension culture adherent culture compared to slower cell growth into a ball, and require at least one week before the massive growth. Once the ball of cells formed to maintain its time is longer than the adherent cultured tumor cells. L9981, A549 cells by the drug cisplatin treatment, resulting in the resistance of cells of various shapes, not size, volume increased significantly, and the translucent difference 3. The adherent cultured L9981 and serum-free suspension culture and serum-containing and Cisplatin induced L9981 doubling time, respectively (46.05 ± 1.95) h and (33.00 ± 1.44) h; adherent A549 and serum-free suspension culture and induced by cisplatin in A549 doubling time, respectively (54.28 ± 1.41) h and (34.49 ± 1.81) h; Serum suspension culture and cis-platinum-induced L9981 cell invasion and into the aneurysm capacity, respectively, is affixed to the wall cultured L9981 cells of 12-fold and 20-fold, with a significant difference (P lt; 0.05); without serum suspension culture and cis-platinum-induced A549 cells invasion ability adherent cultured A549 cells 5 times, with a significant difference (P lt; 0.05) detection of CD133 and CD24 in containing serum stickers wall cultured cells in the expression level, in the absence of serum suspension culture and cisplatin induced processing cell enrichment to the lung cancer stem cell-like cells sub-group, of CD133 expression level significantly significantly improve rather of CD24 expression level significantly with reducing, with a significant difference (P lt; 0.05); nude mice after tumor biopsy confirmed lung cancer, immunohistochemistry results showed that the level of expression of CD133 in which about 2%, CD24 expression was not significant. Conclusion: 1. Preliminary forecast of CD133 CD24-CD133-CD24 may non-small cell lung cancer stem cell surface immune molecules mark, at least close to the mark of lung cancer stem cells. By adding a growth factor in serum-free suspension culture and L9981 and A549 cells induced by cisplatin treatment, Line L9981 by cultured lung cancer spheroids significant effect. L9981 sphere cells, cell proliferation, invasion and tumorigenicity was significantly higher than adherent L9981 cells was statistically significant (P lt; 0.05). A549 into a ball less, serum-free suspension culture and cisplatin treatment of A549 cells induced cell proliferation, invasion and tumorigenicity were significantly higher than the adherent A549 cells was statistically significant (P lt; 0.05). That lung cancer spheroid cell lung cancer stem cell / cancer stem-like cells rich. Lung cancer stem cell-like cell subsets CD133 expression level was significantly increased, and CD24 expression levels were significantly reduced. Surface marker molecules of a certain stage of development prompted CD133 CD24-possible lung cancer stem cell surface marker molecules, or lung cancer stem cells.

Screening of Lung Cancer Stem Cell Surface Markers and Researching Enrichment Methods of Lung Cancer Stem Cell

Abstract

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