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Establishment of a hepatocellular carcinoma cell line with unique metastatic characteristics through in vivo selection and screening for metastasis-related genes through cDNA microarray.
J Cancer Res Clin Oncol. 2003 Jan; 129(1):43-51.JC

Abstract

PURPOSE

To establish a hepatocellular carcinoma (HCC) cell line from lung metastatic lesions of human HCC in nude mice so as to provide a suitable model for the study of lung-metastasis-related molecular mechanisms.

METHODS

HCC clone cells MHCC97-H were inoculated into BALB/c nude mice, and the pulmonary metastatic lesions were harvested and re-implanted into nude mice for the second round of in vivo selection. The same procedure was repeated twice. A new cell line from the third round of lung metastases was established.

RESULTS

A human HCC cell line with unique metastatic characteristics was established by in vivo selection. This cell line, designated as HCCLM3, was polygonal epithelial cell with hypotriploid karyotype and population doubling time of 34.9 h. The cells were positive for alpha fetoprotein (AFP), albumin, cytokeratin 8 (CK8), and negative for hepatitis B surface antigen (HBsAg) by immunocytochemistry. Fluorescence polymerase chain reaction (PCR) showed HBV DNA integration in the cellular genome. When 5 x 10(6) cells were injected subcutaneously into nude mice, tumorigenicity was 100%, with a latency period of 11+/-1 days. Five weeks after s.c. injection, the pulmonary metastatic rate was 100%, the median number of lung metastases being 121 per mouse. After orthotopic implantation of tumor tissue into nude mouse liver for 35 days, widespread loco-regional and distant metastases occurred, with 100% abdominal wall metastases, 80% intra-abdominal cavity metastases, 100% intrahepatic metastases, 70% diaphragm metastases, and 100% pulmonary metastases. The median number of lung metastatic lesions was 268 per mouse. Gene expression profile of HCCLM3 was compared by cDNA microarray with MHCC97-L, a clonal cell strain from the same parental cell line but with low metastatic potential; 25 differentially expressed genes were identified, 18 of which showed decreased expression and seven increased expression in HCCLM3, including the decreased expression of cell cycle control gene Rb2, mismatch repair gene hMSH2, and signal transduction gene protein kinase C beta2, and increased expression of signal transduction gene MAP kinase, kinase 6.

CONCLUSIONS

A new HCC cell line characterized by high pulmonary metastases via s.c. and orthotopic inoculation was established, which provides a new model for the study of liver cancer metastasis. Its gene expression profile could help in the understanding of the mechanism of metastasis and provide potential targets for anti-metastasis intervention.

Authors+Show Affiliations

Liver Cancer Institute and Zhong Shan Hospital of Fudan University, 136 Yi Xue Yuan Road, Shanghai 200032, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12618900

Citation

Li, Y, et al. "Establishment of a Hepatocellular Carcinoma Cell Line With Unique Metastatic Characteristics Through in Vivo Selection and Screening for Metastasis-related Genes Through cDNA Microarray." Journal of Cancer Research and Clinical Oncology, vol. 129, no. 1, 2003, pp. 43-51.
Li Y, Tang Y, Ye L, et al. Establishment of a hepatocellular carcinoma cell line with unique metastatic characteristics through in vivo selection and screening for metastasis-related genes through cDNA microarray. J Cancer Res Clin Oncol. 2003;129(1):43-51.
Li, Y., Tang, Y., Ye, L., Liu, B., Liu, K., Chen, J., & Xue, Q. (2003). Establishment of a hepatocellular carcinoma cell line with unique metastatic characteristics through in vivo selection and screening for metastasis-related genes through cDNA microarray. Journal of Cancer Research and Clinical Oncology, 129(1), 43-51.
Li Y, et al. Establishment of a Hepatocellular Carcinoma Cell Line With Unique Metastatic Characteristics Through in Vivo Selection and Screening for Metastasis-related Genes Through cDNA Microarray. J Cancer Res Clin Oncol. 2003;129(1):43-51. PubMed PMID: 12618900.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Establishment of a hepatocellular carcinoma cell line with unique metastatic characteristics through in vivo selection and screening for metastasis-related genes through cDNA microarray. AU - Li,Y, AU - Tang,Y, AU - Ye,L, AU - Liu,B, AU - Liu,K, AU - Chen,J, AU - Xue,Q, Y1 - 2002/12/04/ PY - 2002/09/21/received PY - 2002/10/28/accepted PY - 2003/3/6/pubmed PY - 2003/5/13/medline PY - 2003/3/6/entrez SP - 43 EP - 51 JF - Journal of cancer research and clinical oncology JO - J Cancer Res Clin Oncol VL - 129 IS - 1 N2 - PURPOSE: To establish a hepatocellular carcinoma (HCC) cell line from lung metastatic lesions of human HCC in nude mice so as to provide a suitable model for the study of lung-metastasis-related molecular mechanisms. METHODS: HCC clone cells MHCC97-H were inoculated into BALB/c nude mice, and the pulmonary metastatic lesions were harvested and re-implanted into nude mice for the second round of in vivo selection. The same procedure was repeated twice. A new cell line from the third round of lung metastases was established. RESULTS: A human HCC cell line with unique metastatic characteristics was established by in vivo selection. This cell line, designated as HCCLM3, was polygonal epithelial cell with hypotriploid karyotype and population doubling time of 34.9 h. The cells were positive for alpha fetoprotein (AFP), albumin, cytokeratin 8 (CK8), and negative for hepatitis B surface antigen (HBsAg) by immunocytochemistry. Fluorescence polymerase chain reaction (PCR) showed HBV DNA integration in the cellular genome. When 5 x 10(6) cells were injected subcutaneously into nude mice, tumorigenicity was 100%, with a latency period of 11+/-1 days. Five weeks after s.c. injection, the pulmonary metastatic rate was 100%, the median number of lung metastases being 121 per mouse. After orthotopic implantation of tumor tissue into nude mouse liver for 35 days, widespread loco-regional and distant metastases occurred, with 100% abdominal wall metastases, 80% intra-abdominal cavity metastases, 100% intrahepatic metastases, 70% diaphragm metastases, and 100% pulmonary metastases. The median number of lung metastatic lesions was 268 per mouse. Gene expression profile of HCCLM3 was compared by cDNA microarray with MHCC97-L, a clonal cell strain from the same parental cell line but with low metastatic potential; 25 differentially expressed genes were identified, 18 of which showed decreased expression and seven increased expression in HCCLM3, including the decreased expression of cell cycle control gene Rb2, mismatch repair gene hMSH2, and signal transduction gene protein kinase C beta2, and increased expression of signal transduction gene MAP kinase, kinase 6. CONCLUSIONS: A new HCC cell line characterized by high pulmonary metastases via s.c. and orthotopic inoculation was established, which provides a new model for the study of liver cancer metastasis. Its gene expression profile could help in the understanding of the mechanism of metastasis and provide potential targets for anti-metastasis intervention. SN - 0171-5216 UR - https://www.unboundmedicine.com/medline/citation/12618900/Establishment_of_a_hepatocellular_carcinoma_cell_line_with_unique_metastatic_characteristics_through_in_vivo_selection_and_screening_for_metastasis_related_genes_through_cDNA_microarray_ L2 - https://dx.doi.org/10.1007/s00432-002-0396-4 DB - PRIME DP - Unbound Medicine ER -