Molecular characterization of invasive subpopulations from an esophageal squamous cell carcinoma cell line.
BACKGROUND
Once diagnosed, esophageal cancer has a very low overall 5-year survival rate. This study investigates the mechanisms behind the invasiveness and severity of esophageal squamous cell carcinoma (ESCC).


METHODS
Transwell invasion chamber was used to subdivide one Taiwanese ESCC cell line, CE81T/VGH, into sublines (CE81T-0, CE81T-1, CE81T-2, CE81T-3, and CE81T-4) in four rounds of assays; the most invasive were identified, and various factors related to their invasiveness measured.


RESULTS
CE81T-1, CE81T-2, CE81T-3 and CE81T-4 sublines were significantly more invasive than the parental cells (CE81T/VGH) and CE81T-0 subline. CE81T-1 and CE81T-4, the sublines we chose to study further, had significantly greater colony-forming ability (3.5- to 2.7-fold) and wound migrating activity (1.95- to 2.6-fold) than the parental cells in vitro (p<0.01). They also displayed greater tumorigenesis in immunodeficient BALB/c Foxlnn mice than the parental cells. We found an inverse correlation between expression of tissue inhibitor of metalloproteinase-2 and invasive ability, and a significant positive correlation between expressions of matrix metalloproteinase-1, vimentin, and p-Src (pY416) in these cell lines and their invasiveness (all p<0.05).


CONCLUSIONS
The subline model may be used to study the molecular and genetic mechanisms underlying the invasion and metastasis of ESCC.
Chen
Yu-Kuei
YK
Department of Family Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan, ROC. 960021@ms.kmuh.org.tw
Chang
Wun-Shaing Wayne
WS
Wu
I-Chen
IC
Li
Ling-Hui
LH
Yang
Sheau-Fang
SF
Chen
Jeff Yi-Fu
JY
Hsu
Ming-Chung
MC
Chen
Shih-Hsien
SH
Wu
Deng-Chyang
DC
Lee
Jang-Ming
JM
Huang
Chun-Hsiung
CH
Goan
Yih-Gang
YG
Chou
Shah-Hwa
SH
Huang
Chia-Tsuan
CT
Wu
Ming-Tsang
MT
D000818:Animals; D015153:Blotting, Western; D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D048708:Cell Growth Processes / Q000502:physiology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D004938:Esophageal Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis / Q000235:genetics; D020778:Matrix Metalloproteinase 2 / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D009361:Neoplasm Invasiveness; D009362:Neoplasm Metastasis; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000096:biosynthesis; D014746:Vimentin / Q000096:biosynthesis / Q000235:genetics
D012333:RNA, Messenger; D014746:Vimentin; D019716:Tissue Inhibitor of Metalloproteinase-2; D020778:Matrix Metalloproteinase 2; D020781:Matrix Metalloproteinase 1