Biologic characteristics of the side population of human small cell lung cancer cell line H446
- VernacularTitle:人小细胞肺癌细胞株H446侧群细胞的生物学特征
- Author:
Wang BO
1
;
Yang HUAN
;
Huang YU-ZHENG
;
Yan RU-HONG
;
Liu FEN-JU
;
Zhang JUN-NING
Author Information
1. 苏州大学附属第一医院
- Keywords:
Small cell lung cancer;
side population;
cancer stem cells;
CD133;
ABCG2
- From:Chinese Journal of Cancer
2010;29(3):272-278
- CountryChina
- Language:Chinese
-
Abstract:
Background and Objective:Recently,the theory of cancer stem cells(CSCs)has presented new targets and orientations for tumor therapy.The major difficulties in researching CSCs include their isolation and purification.The aim of this study is to identify and characterize the side population(SP)cells in small cell lung cancer(SCLC)cell line H446,which Iays the foundation for the isolation and purification of CSCs.Methods:Fluorescence-activated cell sorting(FACS)was used to sort SP and non-SP (NSP)cells from H446,Both subgroups were cultivated to survey the capacity to form into suspended tumor cell spheres.Reverse transcriptionpolymerase chain reaction(RT-PCR)and real-time PCR were used to evaluate the expression levels of the mRNA of CD133,ABCG2,and nucleostemin in both subgroups.The capacity of proliferation and the differences in drug resistance of both subgroups and unso rted cells were tested by the MTT method.The differentiation ability of both subgroups was determined by FACS.Proliferation was determined by subcutaneous tumor formation in nude mice.Results:The percent of Hoechst 33342 negative cells was about(5.1±0.2)%in H446 by fluorescence microscopy.The percent of SP cells was(6.3±0.1)%by flow cytometry.SP cells had a stronger capability of fOrming into tumOr spheres than NSP cells.The mRNA expression Ievels of ABCG2,CD133,and nucleostemin in SP cells were 21.60±0.26,7.10±0.14,and 1.02±0.08 folds higher than that in NSP cells(P＜0.01,P＜0.01,and P＞0.05,respectively).In vivo,SP cells showed better proliferative ability and tougher viability when treated with drugs.SP cells can differentiate into NSP cells,but NSP cells cannot differentiate into SP cells.SP cells had a greater ability to form tumors.Conclusions:The H446 cell line contained some SP cells with stem cell properties.CD133 and ABCG2 may be cancer stem celI markers of SCLC.
