The Application Of OHSV1-hTERT-GFP In The Detection Of Circulating Tumor Cells In Breast Cancer

BackgroundsTumor cells encountered at peripheral blood,that is the circulating tumor cells,have important value in early diagnosis?individual treatment?the evaluation of prognosis.The study is to verify a method to detect circulating tumor cells in the peripheral blood of breast cancer patients based on herpes simplex virus inserted with human telomerase promoter and green fluorescent protein gene(oHS V1-hTERT-GFP).Then,the biological characteristics of tumor cells captured by this method were studied by sequencing.MethodsThe hTERT expression levels of different breast cancer cell lines and leukocytes were detected by Realtime-PCR.Various breast cancer cell lines were infected with oHSV1-hTERT-GFP virus in proportion to MOI=1 to determine the infection rate of this method.Draw about 4ml peripheral blood from healthy people,breast cancer cell lines were mixed with peripheral blood to determine recovery of this method,then Epcam antigen was used to verified the source of the cells.Breast cancer cell lines before and after infected with oHSVl-hTERT-GFP virus were sequenced by transcriptional sequencing to study their biological characteristics.Results:The hTERT were highly expressed in breast cancer cell lines,while leukocytes expressed rarely.The infection rate was higher and the mean recovery rates of MDA-MB-231 was 67.6%.The analysis of transcriptome sequencing of the breast cancer cell lines before and after infection showed that the infection of oHSV1-hTERT-GFP virus changed the biological process obviously.ConclusionsThis study verified the feasibility of a new method for detecting circulating tumor cells.However,the study on the transcriptome level of breast cancer cells infected with oHSV1-hTERT-GFP virus indicates that this method is not suitable for further study of CTCs.