Isolation and Identification of CD133 Positive and Negative Cells from Human Lung Cancer and Screening of the Differential Genes between the Positive and Negative Cells
10.3779/j.issn.1009-3419.2015.03.01
- VernacularTitle:CD133阳性/阴性肺癌细胞的分选、鉴定及差异基因的筛选
- Author:
ZHENG SHAOQIU
1
;
LI SHUHUA
;
WANG HONGYAN
;
XIE XIAOBIN
;
ZHANG YAJIE
Author Information
1. 梅州市人民医院病理科
- Keywords:
CD133;
Adenocarcinoma of lung;
Magnetic activated cell sorting (MACS);
DNA microarray;
Metasta-sis-related genes
- From:
Chinese Journal of Lung Cancer
2015;(3):123-130
- CountryChina
- Language:Chinese
-
Abstract:
Background and objective It has been proven that cancer stem cell existed in variety of cancer, which an significant dierence of biological characteristics was observed between the cancer stem cells and non-cancer stem cells. And CD133 is considered to be cancer stem cell marker. So there may be significant dierences in CD133- positive cells and CD133-negative cells. e aim of this study is to isolate CD133+ cells and CD133- cells from lung cancer cell line A549, ex-plore their biological characteristics and screen the metastasis-related genes. Methods MACS was applied to isolate CD133+cells and CD133- cells from human lung cancer cell line A549. To observe the formation of sphere, CD133+ cells and CD133-cells were cultured in serum-free DMEM-F12 medium (containing EGF, bFGF) in vitro. e colony formaing eciency of CD133+ cells, CD133- cells and cells without sorting was tested by colony-forming assay. e dierentiation of sphere was in-duced by culturing in DMEM-F12 medium (containing serum). e metastasis-related genes (84 genes) of CD133+ cells and CD133- cells were detected by using DNA microarray. Immunohistochemistry was used to detect the expression of CD133 protein in Human lung cancer tissue. Results CD133+ cells formed sphere in serum-free DMEM-F12 medium,while the CD133- cells failed to form sphere. e rates of CD133+ cell colony formation (57.1%) was significantly higher than that of CD133- cells (3.3%). Sphere (CD133+/CK7-) was induced to dierentiate, and CK7 expression was found in dierentiated cells. e expression levels of 19 metastasis-related genes from CD133+ cells and CD133- cells were significant dierent. Lile CD133 positive cells which distributing around the cancer nests were found in lung cancer tissue. e expression of CD133 was not related to tumor types, cell dierentiation or TNM stage. Conclusion CD133+ cells exhibit the characteristics of can-cer stem cells. e dierence of metastasis-related gene expression levels was discovered between CD133+ cells and CD133-cells. CD82 plays an important role in mechanism of tumor metastasis.
