Detection of cancer cells in peripheral blood of breast cancer patients using reverse transcription-polymerase chain reaction for epidermal growth factor receptor.

摘要: The epidermal growth factor receptor (EGFR) has been reported to be expressed in high levels primary breast cancer by immunohistochemistry. In the present study, a reverse transcription (RT)-PCR assay using EGFR primers was developed and evaluated for detection of circulating micrometastases blood patients. Total RNA extracted from cell lines 23 control individuals 37 After transcription, outer nested were used cDNA amplification. integrity confirmed with parallel RT-PCR amplification beta2-microglobulin primers. PCR products electrophoresed on agarose gels containing ethidium bromide visualized UV photography. Southern blotting confirm specificity. capable detecting lower limit 100 fg total A431 line. identified 4 18 (22%) metastatic patients, 0 6 locally recurrent 13 adjuvant controls (P = 0.03, versus control). patients all had progressive disease at time sampling. identity four EGFR-positive bands blotting. presence positivity not treatment-related phenomenon, because three receiving treatment collection. is sensitive specific method proportion cancer.