Establishment and characterization of six human lung cancer cell lines: EGFR, p53 gene mutations and expressions of drug sensitivity genes
Authors
Authors and affiliations
Ja-Lok KuKyung-Hee KimJin-Sung ChoiYou-Kyung JeonSung-Hee KimYoung-Kyoung ShinTae-You KimYung-Jue BangWoo Ho KimJae-Gahb ParkEmail author
1.
2.
3.
Original Paper
First Online: 02 February 2011
425
Downloads
4
Citations
Abstract
Background
Six human lung cancer cell lines (SNU-371, SNU-963, SNU-1327, SNU-1330, SNU-2292 and SNU-2315) were newly established through primary cell cultures. These cell lines were derived from a pulmonary blastoma, a small cell lung cancer, three adenocarcinomas and a squamous cell carcinoma of the lung of six Korean lung cancer patients.
Methods
The histopathology of the primary tumors and their in vitro growth characteristics were described. DNA fingerprinting analysis and genetic alterations in the p53, β-catenin, TGFβRII, K-ras and EGFR genes were conducted. mRNA expressions levels of E-cadherin, COX-2, MDR1, MXR, CGA, synatophysin and TTF1 genes were investigated and sensitivity to anticancer drugs was screened.
Results
Five cell lines grew as adherent cells and one cell line grew as floating aggregates. All lines were free of mycoplasma or bacteria and were proven unique by DNA fingerprinting analysis. A significant polymorphism at codon 72 (Arg to Pro) of the p53 gene was found in one line (SNU-1327) and a mutation at codon 176 was found in SNU-2292. No mutations in the K-ras, β-catenin and TGF-βRII genes were observed. E-cadherin was not expressed in SNU-371 and COX-2 was overexpressed in SNU-1330, SNU-2292 and SNU-2315 cell lines. MDR1 was overexpressed in SNU-371 and SNU-2292 cell lines and MXR was overexpressed in SNU-1327 cell line. Interestingly, the SNU-371 cell line derived from a pulmonary blastoma and which overexpressed MDR1 displayed cross resistance for several anticancer drugs. Neuroendocrine markers, chromogranin A and synaptophysin, were overexpressed in the small cell lung cancer cell line, SNU-963 and thyroid transcription factor-1 was also over expressed in this cell line. Two mutations (p.Glu746_Ser752delinsVal and p.Glu746_Ala750del) in exon 19 of EGFR were found in SNU-1330 and SNU-2315 cell lines, respectively.
Conclusion
These well-characterized lung cancer cell lines may be useful tools for investigations of the biological characteristics of lung cancers, particularly for investigations related to mutations of EGFR.
Keywords
Establishment Lung cancer Cell line EGFR gene
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Notes
Acknowledgments
This work was supported by the S.N.U. foundation & overhead research fund, Korean Cell Line Research Foundation research fund and Priority Research Centers Program National Research Foundation of Korea grant, Ministry of Education, Science and Technology (2009-0093820).
References
1.
A. Jemal, R. Siegel, E. Ward, Y. Hao, J. Xu, T. Murray, M.J. Thun, Cancer statistics, 2008. CA Cancer J. Clin. 58, 71–96 (2008)
PubMedCrossRefGoogle Scholar
2.
H. Shin, K. Jung, Y. Won, J. Park, 2002 Annual Report of the Korea Central Cancer Registry: based on registered data from 139 hospitals. J. Korean Cancer Assoc. 36, 103–114 (2002)
Google Scholar
3.
M. Sanchez-Cespedes, Dissecting the genetic alterations involved in lung carcinogenesis. Lung Cancer 40, 111–121 (2003)
PubMedCrossRefGoogle Scholar
4.
Y.R. Chen, Y.N. Fu, C.H. Lin, S.T. Yang, S.F. Hu, Y.T. Chen, S.F. Tsai, S.F. Huang, Distinctive activation patterns in constitutively active and gefitinib-sensitive EGFR mutants. Oncogene 25, 1205–1215 (2006)
PubMedCrossRefGoogle Scholar
5.
S.W. Han, T.Y. Kim, P.G. Hwang, S. Jeong, J. Kim, I.S. Choi, D.Y. Oh, J.H. Kim, D.W. Kim, D.H. Chung, S.A. Im, Y.T. Kim, J.S. Lee, D.S. Heo, Y.J. Bang, N.K. Kim, Predictive and prognostic impact of epidermal growth factor receptor mutation in non-small-cell lung cancer patients treated with gefitinib. J. Clin. Oncol. 23, 2493–2501 (2005)
PubMedCrossRefGoogle Scholar
6.
A.F. Gazdar, H.K. Oie, Re: Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. Cancer Res. 46, 6011–6012 (1986)
PubMedGoogle Scholar
7.
A.F. Gazdar, H.K. Oie, Cell culture methods for human lung cancer. Cancer Genet. Cytogenet. 19, 5–10 (1986)
PubMedCrossRefGoogle Scholar
8.
J.L. Ku, K.A. Yoon, W.H. Kim, Y. Jang, K.S. Suh, S.W. Kim, Y.H. Park, J.G. Park, Establishment and characterization of four human pancreatic carcinoma cell lines. Genetic alterations in the TGFBR2 gene but not in the MADH4 gene. Cell Tissue Res. 308, 205–214 (2002)
PubMedCrossRefGoogle Scholar
9.
C.S. Koh, J.L. Ku, S.Y. Park, K.H. Kim, J.S. Choi, I.J. Kim, J.H. Park, S.K. Oh, J.K. Chung, J.H. Lee, W.H. Kim, C.W. Kim, B.Y. Cho, J.G. Park, Establishment and characterization of cell lines from three human thyroid carcinomas: responses to all-trans-retinoic acid and mutations in the BRAF gene. Mol. Cell. End
Authors
Authors and affiliations
Ja-Lok KuKyung-Hee KimJin-Sung ChoiYou-Kyung JeonSung-Hee KimYoung-Kyoung ShinTae-You KimYung-Jue BangWoo Ho KimJae-Gahb ParkEmail author
1.
2.
3.
Original Paper
First Online: 02 February 2011
425
Downloads
4
Citations
Abstract
Background
Six human lung cancer cell lines (SNU-371, SNU-963, SNU-1327, SNU-1330, SNU-2292 and SNU-2315) were newly established through primary cell cultures. These cell lines were derived from a pulmonary blastoma, a small cell lung cancer, three adenocarcinomas and a squamous cell carcinoma of the lung of six Korean lung cancer patients.
Methods
The histopathology of the primary tumors and their in vitro growth characteristics were described. DNA fingerprinting analysis and genetic alterations in the p53, β-catenin, TGFβRII, K-ras and EGFR genes were conducted. mRNA expressions levels of E-cadherin, COX-2, MDR1, MXR, CGA, synatophysin and TTF1 genes were investigated and sensitivity to anticancer drugs was screened.
Results
Five cell lines grew as adherent cells and one cell line grew as floating aggregates. All lines were free of mycoplasma or bacteria and were proven unique by DNA fingerprinting analysis. A significant polymorphism at codon 72 (Arg to Pro) of the p53 gene was found in one line (SNU-1327) and a mutation at codon 176 was found in SNU-2292. No mutations in the K-ras, β-catenin and TGF-βRII genes were observed. E-cadherin was not expressed in SNU-371 and COX-2 was overexpressed in SNU-1330, SNU-2292 and SNU-2315 cell lines. MDR1 was overexpressed in SNU-371 and SNU-2292 cell lines and MXR was overexpressed in SNU-1327 cell line. Interestingly, the SNU-371 cell line derived from a pulmonary blastoma and which overexpressed MDR1 displayed cross resistance for several anticancer drugs. Neuroendocrine markers, chromogranin A and synaptophysin, were overexpressed in the small cell lung cancer cell line, SNU-963 and thyroid transcription factor-1 was also over expressed in this cell line. Two mutations (p.Glu746_Ser752delinsVal and p.Glu746_Ala750del) in exon 19 of EGFR were found in SNU-1330 and SNU-2315 cell lines, respectively.
Conclusion
These well-characterized lung cancer cell lines may be useful tools for investigations of the biological characteristics of lung cancers, particularly for investigations related to mutations of EGFR.
Keywords
Establishment Lung cancer Cell line EGFR gene
This is a preview of subscription content, log in to check access
Notes
Acknowledgments
This work was supported by the S.N.U. foundation & overhead research fund, Korean Cell Line Research Foundation research fund and Priority Research Centers Program National Research Foundation of Korea grant, Ministry of Education, Science and Technology (2009-0093820).
References
1.
A. Jemal, R. Siegel, E. Ward, Y. Hao, J. Xu, T. Murray, M.J. Thun, Cancer statistics, 2008. CA Cancer J. Clin. 58, 71–96 (2008)
PubMedCrossRefGoogle Scholar
2.
H. Shin, K. Jung, Y. Won, J. Park, 2002 Annual Report of the Korea Central Cancer Registry: based on registered data from 139 hospitals. J. Korean Cancer Assoc. 36, 103–114 (2002)
Google Scholar
3.
M. Sanchez-Cespedes, Dissecting the genetic alterations involved in lung carcinogenesis. Lung Cancer 40, 111–121 (2003)
PubMedCrossRefGoogle Scholar
4.
Y.R. Chen, Y.N. Fu, C.H. Lin, S.T. Yang, S.F. Hu, Y.T. Chen, S.F. Tsai, S.F. Huang, Distinctive activation patterns in constitutively active and gefitinib-sensitive EGFR mutants. Oncogene 25, 1205–1215 (2006)
PubMedCrossRefGoogle Scholar
5.
S.W. Han, T.Y. Kim, P.G. Hwang, S. Jeong, J. Kim, I.S. Choi, D.Y. Oh, J.H. Kim, D.W. Kim, D.H. Chung, S.A. Im, Y.T. Kim, J.S. Lee, D.S. Heo, Y.J. Bang, N.K. Kim, Predictive and prognostic impact of epidermal growth factor receptor mutation in non-small-cell lung cancer patients treated with gefitinib. J. Clin. Oncol. 23, 2493–2501 (2005)
PubMedCrossRefGoogle Scholar
6.
A.F. Gazdar, H.K. Oie, Re: Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serum-free defined medium. Cancer Res. 46, 6011–6012 (1986)
PubMedGoogle Scholar
7.
A.F. Gazdar, H.K. Oie, Cell culture methods for human lung cancer. Cancer Genet. Cytogenet. 19, 5–10 (1986)
PubMedCrossRefGoogle Scholar
8.
J.L. Ku, K.A. Yoon, W.H. Kim, Y. Jang, K.S. Suh, S.W. Kim, Y.H. Park, J.G. Park, Establishment and characterization of four human pancreatic carcinoma cell lines. Genetic alterations in the TGFBR2 gene but not in the MADH4 gene. Cell Tissue Res. 308, 205–214 (2002)
PubMedCrossRefGoogle Scholar
9.
C.S. Koh, J.L. Ku, S.Y. Park, K.H. Kim, J.S. Choi, I.J. Kim, J.H. Park, S.K. Oh, J.K. Chung, J.H. Lee, W.H. Kim, C.W. Kim, B.Y. Cho, J.G. Park, Establishment and characterization of cell lines from three human thyroid carcinomas: responses to all-trans-retinoic acid and mutations in the BRAF gene. Mol. Cell. End
