Assessment of a Six Gene Panel for the Molecular Detection of Circulating Tumor Cells in the Blood of Female Cancer Patients

Overview

Journal BMC Cancer

Publisher Biomed Central

Specialty Oncology

Date 2010 Dec 7

PMID 21129172

Citations 53

Authors

Eva Obermayr

Fatima Sanchez-Cabo

Muy-Kheng M Tea

Christian F Singer

Michael Krainer

Michael B Fischer

Jalid Sehouli

Alexander Reinthaller

Reinhard Horvat

Georg Heinze

Dan Tong

Robert Zeillinger

Affiliations

Soon will be listed here.

Abstract

Background: The presence of circulating tumor cells (CTC) in the peripheral blood of cancer patients has been described for various solid tumors and their clinical relevance has been shown. CTC detection based on the analysis of epithelial antigens might be hampered by the genetic heterogeneity of the primary tumor and loss of epithelial antigens. Therefore, we aimed to identify new gene markers for the PCR-based detection of CTC in female cancer patients.

Methods: Gene expression of 38 cancer cell lines (breast, ovarian, cervical and endometrial) and of 10 peripheral blood mononuclear cell (PBMC) samples from healthy female donors was measured using microarray technology (Applied Biosystems). Differentially expressed genes were identified using the maxT test and the 50% one-sided trimmed maxT-test. Confirmatory RT-qPCR was performed for 380 gene targets using the AB TaqMan® Low Density Arrays. Then, 93 gene targets were analyzed using the same RT-qPCR platform in tumor tissues of 126 patients with primary breast, ovarian or endometrial cancer. Finally, blood samples from 26 healthy women and from 125 patients (primary breast, ovarian, cervical, or endometrial cancer, and advanced breast cancer) were analyzed following OncoQuick enrichment and RNA pre-amplification. Likewise, hMAM and EpCAM gene expression was analyzed in the blood of breast and ovarian cancer patients. For each gene, a cut-off threshold value was set at three standard deviations from the mean expression level of the healthy controls to identify potential markers for CTC detection.

Results: Six genes were over-expressed in blood samples from 81% of patients with advanced and 29% of patients with primary breast cancer. EpCAM gene expression was detected in 19% and 5% of patients, respectively, whereas hMAM gene expression was observed in the advanced group (39%) only. Multimarker analysis using the new six gene panel positively identified 44% of the cervical, 64% of the endometrial and 19% of the ovarian cancer patients.

Conclusions: The panel of six genes was found superior to EpCAM and hMAM for the detection of circulating tumor cells in the blood of breast cancer, and they may serve as potential markers for CTC derived from endometrial, cervical, and ovarian cancers.

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References

Konigsberg R, Gneist M, Jahn-Kuch D, Pfeiler G, Hager G, Hudec M . Circulating tumor cells in metastatic colorectal cancer: efficacy and feasibility of different enrichment methods. Cancer Lett. 2010; 293(1):117-23. DOI: 10.1016/j.canlet.2010.01.003. View

Muller V, Stahmann N, Riethdorf S, Rau T, Zabel T, Goetz A . Circulating tumor cells in breast cancer: correlation to bone marrow micrometastases, heterogeneous response to systemic therapy and low proliferative activity. Clin Cancer Res. 2005; 11(10):3678-85. DOI: 10.1158/1078-0432.CCR-04-2469. View

Cristofanilli M, Broglio K, Guarneri V, Jackson S, Fritsche H, Islam R . Circulating tumor cells in metastatic breast cancer: biologic staging beyond tumor burden. Clin Breast Cancer. 2007; 7(6):471-9. View

Mikhitarian K, Martin R, Ruppel M, Gillanders W, Hoda R, Schutte D . Detection of mammaglobin mRNA in peripheral blood is associated with high grade breast cancer: interim results of a prospective cohort study. BMC Cancer. 2008; 8:55. PMC: 2292197. DOI: 10.1186/1471-2407-8-55. View

Klein C, Seidl S, Petat-Dutter K, Offner S, Geigl J, Schmidt-Kittler O . Combined transcriptome and genome analysis of single micrometastatic cells. Nat Biotechnol. 2002; 20(4):387-92. DOI: 10.1038/nbt0402-387. View

Cristofanilli M, Hayes D, Budd G, Ellis M, Stopeck A, Reuben J . Circulating tumor cells: a novel prognostic factor for newly diagnosed metastatic breast cancer. J Clin Oncol. 2005; 23(7):1420-30. DOI: 10.1200/JCO.2005.08.140. View

Gentleman R, Carey V, Bates D, Bolstad B, Dettling M, Dudoit S . Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 2004; 5(10):R80. PMC: 545600. DOI: 10.1186/gb-2004-5-10-r80. View

Roncella S, Ferro P, Bacigalupo B, Pronzato P, Tognoni A, Falco E . Human mammaglobin mRNA is a reliable molecular marker for detecting occult breast cancer cells in peripheral blood. J Exp Clin Cancer Res. 2005; 24(2):265-71. View

Forozan F, Karhu R, Kononen J, Kallioniemi A, Kallioniemi O . Genome screening by comparative genomic hybridization. Trends Genet. 1997; 13(10):405-9. DOI: 10.1016/s0168-9525(97)01244-4. View

10.

Gleiss A, Sanchez-Cabo F, Perco P, Tong D, Heinze G . Adaptive trimmed t-statistics for identifying predominantly high expression in a microarray experiment. Stat Med. 2010; 30(1):52-61. DOI: 10.1002/sim.4093. View

11.

Gervasoni A, Monasterio Munoz R, Wengler G, Rizzi A, Zaniboni A, Parolini O . Molecular signature detection of circulating tumor cells using a panel of selected genes. Cancer Lett. 2008; 263(2):267-79. DOI: 10.1016/j.canlet.2008.01.003. View

12.

Cristofanilli M, Budd G, Ellis M, Stopeck A, Matera J, Miller M . Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med. 2004; 351(8):781-91. DOI: 10.1056/NEJMoa040766. View

13.

Marth C, Kisic J, Kaern J, Trope C, Fodstad O . Circulating tumor cells in the peripheral blood and bone marrow of patients with ovarian carcinoma do not predict prognosis. Cancer. 2002; 94(3):707-12. DOI: 10.1002/cncr.10250. View

14.

LAUPER N, Beck A, Cariou S, Richman L, Hofmann K, Reith W . Cyclin E2: a novel CDK2 partner in the late G1 and S phases of the mammalian cell cycle. Oncogene. 1998; 17(20):2637-43. DOI: 10.1038/sj.onc.1202477. View

15.

DIDDLE A, SHOLES Jr D, Hollingsworth J, KINLAW S . Cervical carcinoma; cancer cells in the circulating blood. Am J Obstet Gynecol. 1959; 78:582-5. DOI: 10.1016/0002-9378(59)90530-7. View

16.

Paterlini-Brechot P, Benali N . Circulating tumor cells (CTC) detection: clinical impact and future directions. Cancer Lett. 2007; 253(2):180-204. DOI: 10.1016/j.canlet.2006.12.014. View

17.

Yabushita H, Shimazu M, Yamada H, Sawaguchi K, Noguchi M, Nakanishi M . Occult lymph node metastases detected by cytokeratin immunohistochemistry predict recurrence in node-negative endometrial cancer. Gynecol Oncol. 2001; 80(2):139-44. DOI: 10.1006/gyno.2000.6067. View

18.

Ji X, Sato H, Tanaka H, Konishi Y, Fujimoto T, Takahashi O . Real-time quantitative RT-PCR detection of disseminated endometrial tumor cells in peripheral blood and lymph nodes using the LightCycler System. Gynecol Oncol. 2005; 100(2):355-60. DOI: 10.1016/j.ygyno.2005.08.041. View

19.

Zhong X, Kaul S, Eichler A, Bastert G . Evaluating GA733-2 mRNA as a marker for the detection of micrometastatic breast cancer in peripheral blood and bone marrow. Arch Gynecol Obstet. 2000; 263(1-2):2-6. DOI: 10.1007/s004040050251. View

20.

Stathopoulou A, Mavroudis D, Perraki M, Apostolaki S, Vlachonikolis I, Lianidou E . Molecular detection of cancer cells in the peripheral blood of patients with breast cancer: comparison of CK-19, CEA and maspin as detection markers. Anticancer Res. 2003; 23(2C):1883-90. View