Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97.
OBJECTIVE
To establish clone cells with different metastatic potential for the study of metastasis-related mechanisms.


METHODS
Cloning procedure was performed on parental hepatocellular carcinoma (HCC) cell line MHCC97, and biological characteristics of the target clones selected by in vivo screening were studied.


RESULTS
Two clones with high (MHCC97-H) and low (MHCC97-L) metastatic potential were isolated from the parent cell line. Compared with MHCC97-L, MHCC97-H had smaller cell size (average cell diameter 43 microm vs 50 microm) and faster in vitro and in vivo growth rate (tumor cell doubling time was 34.2h vs 60.0h). The main ranges of chromosomes were 55-58 in MHCC97-H and 57-62 in MHCC97-L. Boyden chamber in vitro invasion assay demonstrated that the number of penetrating cells through the artificial basement membrane was (37.5 +/- 11.0) cells/field for MHCC97-H vs (17.7 +/- 6.3)/field for MHCC97-L. The proportions of cells in G0-G1 phase, S phase, and G2-M phase for MHCC97-H/MHCC97-L were 0.56/0.65, 0.28/0.25 and 0.16/0.10, respectively, as measured by flow cytometry. The serum AFP levels in nude mice 5wk after orthotopic implantation of tumor tissue were (246 +/- 66) microg.L(-1) for MHCC97-H and (91 +/- 66) microg.L(-1) for MHCC97-L. The pulmonary metastatic rate was 100% (10/10) vs 40% (4/10).


CONCLUSIONS
Two clones of the same genetic background but with different biological behaviors were established, which could be valuable models for investigation on HCC metastasis.
Li
Y
Y
Liver Cancer Institute & Zhongshan Hospital of Fudan University, 136 Yixueyuan Road, Shanghai 200032,China.
Tang
Z Y
ZY
Ye
S L
SL
Liu
Y K
YK
Chen
J
J
Xue
Q
Q
Chen
J
J
Gao
D M
DM
Bao
W H
WH
D000418:Albumins / Q000032:analysis; D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000556:secondary*; D002455:Cell Division; D002875:Chromosomes; D002999:Clone Cells; D005434:Flow Cytometry; D006509:Hepatitis B / Q000473:pathology; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000302:isolation & purification; D006801:Humans; D007633:Keratins / Q000032:analysis; D008099:Liver / Q000473:pathology*; D008114:Liver Neoplasms, Experimental / Q000235:genetics / Q000556:secondary*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000166:cytology / Q000821:virology; D016662:Virus Integration; D000509:alpha-Fetoproteins / Q000032:analysis
D000418:Albumins; D006514:Hepatitis B Surface Antigens; D000509:alpha-Fetoproteins; D007633:Keratins