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Gene expression of somatostatin receptor subtypes SSTR2a, SSTR3 and SSTR5 in peripheral blood of neuroendocrine lung cancer affected patients

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Abstract

Background

Somatostatin (SS) acts as a universal endocrine off-switch, and also inhibits the growth of neuroendocrine tumours through its specific receptors (SSTRs). Somatostatin receptors are G-protein-coupled receptors, which are encoded by five separate genes (SSTR1-5). Short peptide analogues demonstrate specific binding only for the subgroup consisting of SSTR2a, SSTR3 and SSTR5. Moreover, previous studies reported that expression of mRNA for SSTR2a correlated with therapeutic outcome in patients with carcinoid tumours treated with somatostatin analogs.

Purpose

To develop and apply a Real Time Quantitative PCR technique (RT-qPCR) to compare and contrast the mRNA levels of SSTR2a, SSTR3 and SSTR5 in Neuroendocrine Lung Cancer affected patients.

Methods

Peripheral blood samples from 21 neuroendocrine lung cancer affected patients (14 SCLC, 6 LC and 1 LCNEC) subjected to scintigraphy with 111In-DTPA-D-Phe1-octreotide (OctreoScan) and 24 healthy blood donors were investigated by RT-qPCR. mRNA levels for SSTR2a, SSTR3 and SSTR5 were measured in peripheral blood samples with a relative quantification method using plasmid dilutions as calibration curves and GAPDH as reference gene.

Results

A statistically significant increase in target genes/GAPDH copy number ratio was found for SSTR2a (median 38; IQR 22–141) and SSTR5 (median 51; IQR 19–499) in neuroendocrine lung cancer affected patients as compared with samples from healthy blood donors (P ≤ 0.0003 and P ≤ 0.0005). Since low levels of expression were detected in the control group for all three genes, optimal cut-off values were assessed using ROC curve analyses and were equal to 9.05 for SSTR2a and 16.97 for SSTR5. These cut off values resulted in a sensitivity of 86% (95%IC 65–95) for both markers and a specificity of 83% (95%IC 64–93%) and 79% (95%IC 60–91%) for SSTR2a and SSTR5 respectively. Comparison between OctreoScan results and RT-qPCR analysis demonstrated agreement in 76% of the cases.

Conclusions

Our results suggest that SSTR2a and SSTR5 mRNAs are detectable in peripheral blood of neuroendocrine lung cancer affected patients using real-time quantitative PCR, with a good agreement with OctreoScan. The high sensitivity of this non-invasive molecular technique suggests that this method could represent a useful tool in the clinical management of neuroendocrine lung cancers.

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Acknowledgements

The authors wish to thank the patients and the volunteers for their indispensable co-operation. This study was supported by the Italian Ministry of Health (Ricerca Corrente 2010 RC1003MG58). The authors declare no potential conflicts of interest.

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Authors and Affiliations

  1. Laboratory of Oncology, IRCCS “Casa Sollievo della Sofferenza” Hospital, San Giovanni Rotondo, Foggia, Italy

    Lucia Anna Muscarella, Annamaria la Torre, Paola Parrella & Vito Michele Fazio

  2. Internal Medicine Unit, IRCCS “Casa Sollievo della Sofferenza” Hospital, San Giovanni Rotondo, Foggia, Italy

    Vito D’Alessandro, Angelo De Cata, Marco Sperandeo & Gianluigi Vendemiale

  3. Unit of Biostatistics, IRCCS “Casa Sollievo della Sofferenza” Hospital, San Giovanni Rotondo, Foggia, Italy

    Massimiliano Copetti & Fabio Pellegrini

  4. Nuclear Medicine Unit, IRCCS “Casa Sollievo della Sofferenza” Hospital, San Giovanni Rotondo, Foggia, Italy

    Vincenzo Frusciante

  5. Oncology Unit, IRCCS “Casa Sollievo della Sofferenza” Hospital, San Giovanni Rotondo, Foggia, Italy

    Evaristo Maiello

  6. Medical Genetics Unit, IRCCS “Casa Sollievo della Sofferenza” Hospital, San Giovanni Rotondo, Foggia, Italy

    Giuseppe Merla

  7. Department of Clinical Pharmacology and Epidemiology, Consorzio Mario Negri Sud, Santa Maria Imbaro, Chieti, Italy

    Fabio Pellegrini

Authors
  1. Lucia Anna Muscarella
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  2. Vito D’Alessandro
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  3. Annamaria la Torre
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  4. Massimiliano Copetti
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  9. Vincenzo Frusciante
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  10. Evaristo Maiello
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  11. Giuseppe Merla
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  12. Vito Michele Fazio
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  13. Gianluigi Vendemiale
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Corresponding authors

Correspondence to Lucia Anna Muscarella or Vito D’Alessandro.

Additional information

Lucia Anna Muscarella, Vito D’Alessandro, Vito Michele Fazio, Gianluigi Vendemiale have equally contributed to this work

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Supplemental Table 1

SSTR marker expression also for the different subcategories of lung cancer patients. (DOC 69.5 kb)

Supplemental Figure 1

Standard curves for SSTR2a, SSTR3 and SSTR5 plasmid dilutions. The graph shows the mean of Ct values versus log of plasmid copy numbers, measured in triplicate. Slope and intercept are represented in the equations of the regression lines, together with regression coefficient. (DOC 82 kb)

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Muscarella, L.A., D’Alessandro, V., la Torre, A. et al. Gene expression of somatostatin receptor subtypes SSTR2a, SSTR3 and SSTR5 in peripheral blood of neuroendocrine lung cancer affected patients. Cell Oncol. 34, 435–441 (2011). https://doi.org/10.1007/s13402-011-0025-9

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