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Background  Leucine-rich repeats and immunoglobulin-like domains 1 ( LRIG1 ) copy number alterations and unbalanced gene recombination events have been reported to occur in breast cancer. Importantly,  LRIG1  loss was recently shown to predict early and late relapse in stage I-II breast cancer.    Methods  We developed droplet digital PCR (ddPCR) assays for the determination of relative  LRIG1  copy numbers and used these assays to analyze  LRIG1  in twelve healthy individuals, 34 breast tumor samples previously analyzed by fluorescence in situ hybridization (FISH), and 423 breast tumor cytosols.    Results  Four of the  LRIG1 /reference gene assays were found to be precise and robust, showing copy number ratios close to 1 (mean, 0.984; standard deviation, +/− 0.031) among the healthy control population. The correlation between the ddPCR assays and previous FISH results was low, possibly because of the different normalization strategies used. One in 34 breast tumors (2.9%) showed an unbalanced  LRIG1  recombination event.  LRIG1  copy number ratios were associated with the breast cancer subtype, steroid receptor status,  ERBB2  status, tumor grade, and nodal status. Both  LRIG1  loss and gain were associated with unfavorable metastasis-free survival; however, they did not remain significant prognostic factors after adjustment for common risk factors in the Cox regression analysis. Furthermore,  LRIG1  loss was not significantly associated with survival in stage I and II cases.    Conclusions  Although  LRIG1  gene aberrations may be important determinants of breast cancer biology, and prognostic markers, the results of this study do not verify an important role for  LRIG1  copy number analyses in predicting the risk of relapse in early-stage breast cancer.

LRIG1 gene copy number analysis by ddPCR and correlations to clinical factors in breast cancer