ALC  Vol.2 No.1 , March 2013
Behavior of circulating epithelial tumor cells (CETC) and FISH (fluorescence in situ hybridisation) of epidermal growth factor receptor (EGFR)-gene amplification in lung cancer patients during the course of therapy
Abstract: Introduction: Monitoring the response of CETC to therapy in lung cancer allows early detection of patients at risk of progression. Analysis of the EGFR-gene amplification in these cells may help to characterize patients who might benefit from tyrosine kinase inhibitors. Methods: CETCs were quantified at least twice during treatment from blood of 52 patients with advanced non small cell lung cancer (NSCLC) using fluorescence labelled anti-EpCAM. EGFR-gene amplification was analysed in these cells with double probe (EGFR/CEP7) using FISH analysis. Results: Progression of the tumor was observed in 30 of the 52 patients (58%). With respect to changes in CETCs during therapy and progression free survival 31 patients showed a decrease in CETCs, 2 developing a single brain metastasis and 12 progressive disease; 20 patients showed an increase in CETC more than twofold 16 of which developed progressive disease. The difference was highly significant (p=0.007 Fisher’s exact test) irrespective of age, sex, tumor size, pathological type and therapy. Kaplan-Meier progression free survival was significantly different between patients with decreasing and increaseing CETC (p=0.038). 5/20 patients tested were positive for EGFR amplification with 85-100% of EpCAM positive cells showing this chromosomal abnormality. One patient could be followed during therapy with increasing CETC during therapy with bevacizumab followed by relapse. He subsequently received erlotinib resulting in a decrease in CETC and is still free of progress after 516 days. Conclusions: These results show that peripherally circulating tumor cells in patients with advanced NSCLC are influenced by systemic chemotherapy and an increase in spite of therapy is a marker of aggressiveness of the tumor cells. Determination of the EGFR amplification might help to better treat part of these patients.
Cite this paper: El Sherif, M., Schneider, C., Rabenstein, C., Hassab, A., El Bordiny, M., Ayad, M. and Pachmann, K. (2013) Behavior of circulating epithelial tumor cells (CETC) and FISH (fluorescence in situ hybridisation) of epidermal growth factor receptor (EGFR)-gene amplification in lung cancer patients during the course of therapy. Advances in Lung Cancer, 2, 1-8. doi: 10.4236/alc.2013.21001.

[1]   Jemal, A., Thun, M.J., Ries, L.A., Howe, H.L., Weir, H.K., Center, M.M., Ward, E., Wu, X.C., Eheman, C., Anderson, R., Ajani, U.A., Kohler, B. and Edwards, B.K., (2008) Annual report to the nation on the status of cancer, 1975-2005, featuring trends in lung cancer, tobacco use, and tobacco control. Journal of the National Cancer Initute, 100, 1672-1694. doi:10.1093/jnci/djn389

[2]   Jemal, A., Murray, T., Sammuels, A., Ghafoor, A., Ward, E. and Thun, M.J., (2003) Cancer statistics. A Cancer Journal for Clinicians, 53, 5-26.

[3]   Chen, Y.C., Hsu, H.S., Chen, Y.W., Tsai, T.H., How, C.K., Wang, C.Y., Hung, S.C., Chang, Y.L., Tsai, M.L., Lee, Y.Y., Ku, H.H. and Chiou, S.H., (2008) Oct 4 expression maintained cancer stem like properties in lung cancer-erived CD133-positive cells. Plosone, 3, e2637. doi:10.1371/journal.pone.0002637

[4]   Borczuk, A.C., Gorenstein, L., Walter, K.L., Assaad, A.A. and Wang, L., (2003) Non-smal-cell lung cancer molecular signatures recapitulate lung developmental pathways. American Journal of Pathology, 163, 1949-1960. doi:10.1016/S0002-9440(10)63553-5

[5]   Ashworth, T.R., (1969) A case of cancer in which similar cells similar to those in the tumour were seen in the blood after death. Australian Medical Journal, 14, 146-149.

[6]   Liotta, L.A., KLeinerman, J. and Saidel, G.M. (1974) Quantitative relationship of intravascular tumor cells, tumor vessels and pulmonary metastases following tumor implantation. Cancer Research, 34, 997-1004.

[7]   Meng, S., Tripathy, D., Frenkel, E.P., Shete, S., Naftalis, E.Z., Huth, J.F., Beitsch, P.D., Leitch, M., Hoover, S., Euhus, D., Haley, B., Morrison, L., Fleming, T.P., Herlyn, D., Terstappen, L.W., Fehm, T., Tucker, T.F., Lane, N., Wang, J. and Uhr, J.W., (2004) Circulating tumor cells in patients with breast cancer dormancy. Clinical Cancer Research, 10, 8152-8162. doi:10.1158/1078-0432.CCR-04-1110

[8]   Pachmann, K., (2005) Long time recirculating tumor cells in breast cancer patients. Clinical Cancer Research, 11, 5657-5658. doi:10.1158/1078-0432.CCR-05-0191

[9]   Pachmann, K., Clement, J.H., Schneider, C.P., Willen, B., Camara, O., Pachmann, U. and H?ffken, K., (2005) Standardized quantification of circulating peripheral tumor cells from lung and breast cancer. Clinical Chemistry and Laboratory Medicine, 43, 617-627. doi:10.1515/CCLM.2005.107

[10]   Pachmann, K., Camara, O., Kavallaris, A., Schneider, U., Schünemann, S. and H?ffken, K., (2005) Quantification of the response of circulating epithelial cells to neoadjuvant treatment for breast cancer: A new tool for therapy monitoring. Breast Cancer Research, 7, 975-979. doi:10.1186/bcr1328

[11]   Solomayer, E.F., Diel, I.J., Salanti, G., Hahn, M., Gollan, C., Schütz, F. and Bastert, G., (2001) Time independence of the prognostic impact of tumour cell detection in the BM of primary breast cancer patients. Clinical Cancer Research, 7, 4102-4108.

[12]   Mansi, J.L., Gogas, H., Bliss, J.M., Gazet, J.C., Berger, U. and Coombes, R.C., (1999) Outcome of primary breast cancer patients with micrometastasis: A long term follow up study. Lancet, 354, 197-202. doi:10.1016/S0140-6736(98)10175-7

[13]   Janni, W., Rack, B., Lindemann, K. and Harbeck, N. (2005) Detection of micrometastatic disease in bone mar- row: Is it ready for prime time? Oncologist, 10, 480-492. doi:10.1634/theoncologist.10-7-480

[14]   Rolle, A., Günzel, R., Pachmann, U., Willen, B., H?ffken, K. and Pachmann, K. (2005) Increase in number of circulating disseminated epithelial cells after surgery for non-small lung cancer monitored by Maintrac is a predictor for relapse: A preliminary report. World Journal of Surgical Ooncology, 3, 18. doi:10.1186/1477-7819-3-18

[15]   Camara, O., Kavallaris, A., N?schel, H., Rengsberger, M., J?rke, C. and Pachmann, K., (2006) Seeding of epithelial cells into circulation during surgery for breast cancer: The fate of malignant and benign mobilized cells. World Journal of Surgical Ooncology, 4, 67. doi:10.1186/1477-7819-4-67

[16]   Anni, W., Rack, B., Schindlbeck, C., Strobl, B., Rjosk, D., Braun, S., Sommer, H., Pantel, K., Gerber, B. and Friese, K. (2005) The persistence of isolated tumour cells in bone marrow from patients with breast carcinoma predicts an increase for recurrence. Cancer, 103, 884-891. doi:10.1002/cncr.20834

[17]   Müller, V., Stahmann, N., Riethdorf, S., Rau, T., Zabel, T., Goetz, A., J?nicke, F. and Pantel, K., (2005) Circulating tumor cells in breast cancer: Correlation to bone marrow micrometastases, heterogeneous response to systemic therapy and low proliferative activity. Clinical Cancer Research, 11, 3678-3685. doi:10.1158/1078-0432.CCR-04-2469

[18]   Vlems, F.A., Ladanyi, A., Gertler, R., Rosenberg, R., Diepstra, J.H., R?der, C., Nekarda, H., Molnar, B., Tulassay, Z., van Muijen, G.N. and Vogel, I., (2003) Reliability of quantitative reverse-transcriptase-PCR-based detection of tumour cells in the blood between different laboratories using a standardised protocol. European Journal of Cancer, 39, 388-396. doi:10.1016/S0959-8049(02)00631-7

[19]   Baran, J., Pituch-Noworolska, A., Krzeszowiak, A., Wieckiewicz, J, Stachura, J., Popiela, T., Szczepanik, A. and Zembala, M., (1998) Detection of cancer cells in the blood by FACS sorting of CD 45-cells. International Journal of Molecular Medicine, 1, 573-578.

[20]   Ntouroupi, T.G., Ashraf, S.Q., McGregor, S.B., Turney, B.W., Seppo, A., Kim, Y., Wang, X., Kilpatrick, M.W., Tsipouras, P., Tafas, T. and Bodmer, W.F., (2008) Detection of circulating tumour cells in peripheral blood with an automated scanning fluorescence microscope. British Journal of Cancer, 99, 789. doi:10.1038/sj.bjc.6604545

[21]   Hirsch, F.R., Varella-Garcia, M., Bunn Jr, P.A., Di Maria, M.V., Veve, R., Bremmes, R.M., Barón, A.E., Zeng, C. and Franklin, W.A. (2003) Epidermal growth factor receptor in non-small-cell lung carcinoma: Correlation be- tween gene copy number and protein expression and impact on prognosis. Journal of Clinical Oncology, 21, 3798-3807. doi:10.1200/JCO.2003.11.069

[22]   Shepherd, F.A., Rodrigues, Pereira, J., Ciuleanu, T., Tan, E.H., Hirsh, V., Thongprasert, S., Campos D., Maoleekoonpiroj, S., Smylie, M., Martins, R., van Kooten, M., Dediu, M., Findlay, B., Tu D., Johnston, D., Bezjak A., Clark, G, Santabárbara, P., Seymour, L. and National Cancer Institute of Canada Clinical Trials Group, (2005) Erlotinib in previously treated non-small-cell lung cancer. New England Journal of Medicine, 353, 123-132. doi:10.1056/NEJMoa050753

[23]   Morinaga, R., Okamoto, I., Fujita, Y., Arao, T., Sekijima, M., Nishio, K., Ito, H., Fukuoka, M., Kadota, J. and Nakagawa, K., (2008) Association of epidermal growth factor receptor (EGFR) gene mutations with EGFR amplifycations in advanced non-small cell lung cancer. Cancer Science, 99, 2455-2460. doi:10.1111/j.1349-7006.2008.00962.x

[24]   Pachmann, K., Camara O., Kohlhase, A., Rabenstein, C., Kroll, T., Runnebaum, I.B. and Hoeffken, K., (2010) Assessing the efficacy of targeted therapy using circulating epithelial tumor cells (CETC): The example of SERM therapy monitoring as a unique tool to individualise therapy. Journal of Cancer Research and Clinical Oncology, 137, 821-828. doi:10.1007/s00432-010-0942-4

[25]   Lin, H., Balic, M., Zheng, S., Datar, R. and Cote, R.J. (2011) Disseminated and circulating tumor cells: Role in effective cancer management. Critical Reviews in Oncology/Hematology, 77, 1-11. doi:10.1016/j.critrevonc.2010.04.008

[26]   Pachmann, K., Oumar, C., Kavallaris, A., Sabine, K., Malarski, N., Gajda, M., Kroll, T., J?rke, C., Hammer, U., Altendorf-Hofmann, A., Rabenstein, C., Pachmann, U., Runnebaum, I. and H?ffken, A., (2008) Monitoring the response of circulating epithelial tumor cells to adjuvant chemotherapy in breast cancer allows detection of patients at risk of early relapse. Journal of Clinical Oncology, 26, 1208-1215. doi:10.1200/JCO.2007.13.6523

[27]   Camara, O., Rengsberger, M., Egbe, A., Koch, A., Gajda, M., Hammer, U., Rabenstein, C., Untch, M. and Pachmann, K., (2007) The relevance of circulating epithelial tumor cells (CETC) for therapy monitoring during neoadjuvant (primary systemic) chemotherapy in breast cancer. Annals of Oncology, 18, 1484-1492. doi:10.1093/annonc/mdm206

[28]   Maheswaran, S., Sequist, L.V., Nagrath, S., Ulkus, L., Brannigan, B., Collura, C.V., Inserra, E., Diederichs, S., Iafrate, A.J., Bell, D.W., Digumarthy, S., Muzikansky, A., Irimia, D., Settleman, J., Tompkins R.G., Lynch, T.J., Toner, M. and Haber, D.A., (2008) Detection of mutations in EGFR in circulating lung-cancer cells. New England Journal of Medicine, 359, 366-377. doi:10.1056/NEJMoa0800668

[29]   Sequist, L.V., Martins, R.G., Spigel, D., Grunberg, S.M., Spira, A., J?nne, P.A., Joshi, V.A., McCollum, D., Evans, T.L., Muzikansky, A., Kuhlmann, G.L., Han, M., Goldberg, J.S., Settleman, J., Iafrate, A.J., Engelman J.A., Haber, D.A., Johnson, B.E. and Lynch, T.J., (2008) First-line gefitinib in advanced nonsmall-cell lung cancer patients harboring somatic EGFR mutations. Journal of Clinical Oncology, 26, 2442-2449. doi:10.1200/JCO.2007.14.8494

[30]   Therasse, P., Arbuck, S.G., Eisenhauer, E.A., Wanders, J., Kaplan, R.S., Rubinstein, L., Verweij, J., Van Glabbeke, M., van Oosterom, A.T., Christian, M.C. and Gwyther, S.G., (2000) New guidelines to evaluate the response to treatment in solid tumors. European Organization for Research and Treatment of Cancer, National Cancer Institute of the United States, National Cancer Institute of Canada. Journal of National Cancer Institute, 92, 205-216. doi:10.1093/jnci/92.3.205