Limits...
Establishment and characterization of a novel primary hepatocellular carcinoma cell line with metastatic ability in vivo.

Cheung PF, Yip CW, Ng LW, Lo KW, Wong N, Choy KW, Chow C, Chan KF, Cheung TT, Poon RT, Fan ST, Cheung ST - Cancer Cell Int. (2014)

Bottom Line: The cell line was authenticated by short tandem repeat analysis.Sequence analysis revealed p53 gene mutation.The newly established cell line could serve as useful in vitro and in vivo models for studying primary HCC that possess metastasis ability.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, The University of Hong Kong, Hong Kong, China ; Center for Cancer Research, The University of Hong Kong, Hong Kong, China.

ABSTRACT

Background: Hepatocellular carcinoma (HCC) is a highly aggressive and heterogeneous disease. HCC cell lines established from different patients would be useful in elucidating the molecular pathogenesis. However, success of HCC primary culture establishment remains at low rate. We aim to establish and characterize HCC primary culture and the derived cell line.

Methods: Fresh tumor tissues were collected from 30 HCC patients. Culture conditions were optimized for the attachment and growth of the isolated hepatocytes. Granulin-epithelin precursor (GEP), a growth factor reported to associate with cancer stem cell properties, was examined by flow cytometry to elucidate its role on primary culture establishment. The primary cell line was characterized in detail.

Results: Cells isolated from 16 out of 30 HCC cases (53%) had viability more than 70% and were subject to subsequent in vitro culture. 7 out of 16 cases (44%) could give rise to cells that were able to attach and grow in culture. GEP expression levels significantly correlated with the viability of isolated hepatocytes and success rate of subsequent primary culture establishment. Cells from HCC patient 21 grew and expanded rapidly in vitro and was selected to be further characterized. The line, designated HCC21, derived from a Hong Kong Chinese female patient with HCC at Stage II. The cells exhibited typical epithelial morphology and expressed albumin, AFP and HBV antigens. The cell line was authenticated by short tandem repeat analysis. Comparative genome hybridization analysis revealed chromosomal loss at 1p35-p36, 1q44, 2q11.2-q24.3, 2q37, 4q12-q13.3, 4q21.21-q35.2, 8p12-p23, 15q11.2-q14, 15q24-q26, 16p12.1-p13.3, 16q, 17p, 22q and gain at 1q21-q43 in both HCC21 cells and the original clinical tumor specimen. Sequence analysis revealed p53 gene mutation. Subcutaneous injection of HCC21 cells into immunodeficient mice showed that the cells were able to form tumors at the primary injection sites and metastatic tumors in the peritoneal cavity.

Conclusions: The newly established cell line could serve as useful in vitro and in vivo models for studying primary HCC that possess metastasis ability.

No MeSH data available.


Related in: MedlinePlus

CGH analysis of HCC21 cells and the original tumor. Vertical profiles of chromosomes 1, 2, 4, 8, 15, 16, 17 and 22 are shown with a log2 ratio, which represents the difference between tumor and normal DNA copy number. Ratio of 0 indicates that there is no change in copy number between tumor and normal cell DNA. Positive and negative log2 ratios indicate the gain and loss of tumor DNA copy number, respectively. The left panel of each plot represents the chromosome of the original clinical tumor specimen, while the right panel represents that of HCC21 cells at passage 10.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4209051&req=5

Fig5: CGH analysis of HCC21 cells and the original tumor. Vertical profiles of chromosomes 1, 2, 4, 8, 15, 16, 17 and 22 are shown with a log2 ratio, which represents the difference between tumor and normal DNA copy number. Ratio of 0 indicates that there is no change in copy number between tumor and normal cell DNA. Positive and negative log2 ratios indicate the gain and loss of tumor DNA copy number, respectively. The left panel of each plot represents the chromosome of the original clinical tumor specimen, while the right panel represents that of HCC21 cells at passage 10.

Mentions: Changes in genomic copy number in HCC21 cells and original tumor were characterized by aCGH analysis. aCGH arrays were performed using DNA from original tumor and that from the HCC21 cells 6 months after its establishment. Chromosomal aberrations including chromosomal loss at 1p35-p36, 1q44, 2q11.2-q24.3, 2q37, 4q12-q13.3, 4q21.21-q35.2, 8p12-p23, 15q11.2-q14, 15q24-q26, 16p12.1-p13.3, 16q, 17p, 22q and gain at 1q21-q43 were observed in both the original tumor specimen and HCC21 cells (Figure 5). Noted the STR profiles showed LOH at FGA (at chromosome 4q28) and D16S539 (at chromosome 16q24.1), and that corroborated the aCGH data with chromosomal loss at 4q21.21-q35.2 and 16q in both the original clinical tumor specimen and the derived HCC21 cells.Figure 5


Establishment and characterization of a novel primary hepatocellular carcinoma cell line with metastatic ability in vivo.

Cheung PF, Yip CW, Ng LW, Lo KW, Wong N, Choy KW, Chow C, Chan KF, Cheung TT, Poon RT, Fan ST, Cheung ST - Cancer Cell Int. (2014)

CGH analysis of HCC21 cells and the original tumor. Vertical profiles of chromosomes 1, 2, 4, 8, 15, 16, 17 and 22 are shown with a log2 ratio, which represents the difference between tumor and normal DNA copy number. Ratio of 0 indicates that there is no change in copy number between tumor and normal cell DNA. Positive and negative log2 ratios indicate the gain and loss of tumor DNA copy number, respectively. The left panel of each plot represents the chromosome of the original clinical tumor specimen, while the right panel represents that of HCC21 cells at passage 10.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4209051&req=5

Fig5: CGH analysis of HCC21 cells and the original tumor. Vertical profiles of chromosomes 1, 2, 4, 8, 15, 16, 17 and 22 are shown with a log2 ratio, which represents the difference between tumor and normal DNA copy number. Ratio of 0 indicates that there is no change in copy number between tumor and normal cell DNA. Positive and negative log2 ratios indicate the gain and loss of tumor DNA copy number, respectively. The left panel of each plot represents the chromosome of the original clinical tumor specimen, while the right panel represents that of HCC21 cells at passage 10.
Mentions: Changes in genomic copy number in HCC21 cells and original tumor were characterized by aCGH analysis. aCGH arrays were performed using DNA from original tumor and that from the HCC21 cells 6 months after its establishment. Chromosomal aberrations including chromosomal loss at 1p35-p36, 1q44, 2q11.2-q24.3, 2q37, 4q12-q13.3, 4q21.21-q35.2, 8p12-p23, 15q11.2-q14, 15q24-q26, 16p12.1-p13.3, 16q, 17p, 22q and gain at 1q21-q43 were observed in both the original tumor specimen and HCC21 cells (Figure 5). Noted the STR profiles showed LOH at FGA (at chromosome 4q28) and D16S539 (at chromosome 16q24.1), and that corroborated the aCGH data with chromosomal loss at 4q21.21-q35.2 and 16q in both the original clinical tumor specimen and the derived HCC21 cells.Figure 5

Bottom Line: The cell line was authenticated by short tandem repeat analysis.Sequence analysis revealed p53 gene mutation.The newly established cell line could serve as useful in vitro and in vivo models for studying primary HCC that possess metastasis ability.

View Article: PubMed Central - PubMed

Affiliation: Department of Surgery, The University of Hong Kong, Hong Kong, China ; Center for Cancer Research, The University of Hong Kong, Hong Kong, China.

ABSTRACT

Background: Hepatocellular carcinoma (HCC) is a highly aggressive and heterogeneous disease. HCC cell lines established from different patients would be useful in elucidating the molecular pathogenesis. However, success of HCC primary culture establishment remains at low rate. We aim to establish and characterize HCC primary culture and the derived cell line.

Methods: Fresh tumor tissues were collected from 30 HCC patients. Culture conditions were optimized for the attachment and growth of the isolated hepatocytes. Granulin-epithelin precursor (GEP), a growth factor reported to associate with cancer stem cell properties, was examined by flow cytometry to elucidate its role on primary culture establishment. The primary cell line was characterized in detail.

Results: Cells isolated from 16 out of 30 HCC cases (53%) had viability more than 70% and were subject to subsequent in vitro culture. 7 out of 16 cases (44%) could give rise to cells that were able to attach and grow in culture. GEP expression levels significantly correlated with the viability of isolated hepatocytes and success rate of subsequent primary culture establishment. Cells from HCC patient 21 grew and expanded rapidly in vitro and was selected to be further characterized. The line, designated HCC21, derived from a Hong Kong Chinese female patient with HCC at Stage II. The cells exhibited typical epithelial morphology and expressed albumin, AFP and HBV antigens. The cell line was authenticated by short tandem repeat analysis. Comparative genome hybridization analysis revealed chromosomal loss at 1p35-p36, 1q44, 2q11.2-q24.3, 2q37, 4q12-q13.3, 4q21.21-q35.2, 8p12-p23, 15q11.2-q14, 15q24-q26, 16p12.1-p13.3, 16q, 17p, 22q and gain at 1q21-q43 in both HCC21 cells and the original clinical tumor specimen. Sequence analysis revealed p53 gene mutation. Subcutaneous injection of HCC21 cells into immunodeficient mice showed that the cells were able to form tumors at the primary injection sites and metastatic tumors in the peritoneal cavity.

Conclusions: The newly established cell line could serve as useful in vitro and in vivo models for studying primary HCC that possess metastasis ability.

No MeSH data available.


Related in: MedlinePlus