title: Genomic DNA of MCF-7 breast cancer cells not an ideal choice as positive control for PCR amplification based detection of Mouse Mammary Tumor Virus-Like Sequences.
abstract: The identification of the etiology of breast cancer is a crucial research issue for the development of an effective preventive and treatment strategies. Researchers are exploring the possible involvement of Mouse Mammary Tumor Virus (MMTV) in causing human breast cancer. Hence, it becomes very important to use a consistent positive control agent in PCR amplification based detection of MMTV-Like Sequence (MMTV-LS) in human breast cancer for accurate and reproducible results. This study was done to investigate the feasibility of using genomic DNA of MCF-7 breast cancer cells to detect MMTV-LS using PCR amplification based detection. MMTV env and SAG gene located at the 3' long terminal repeat (LTR) sequences were targeted for the PCR based detection. No amplification was observed in case of the genomic DNA of MCF-7 breast cancer cells. However, the 2.7 kb DNA fragment comprising MMTV env and SAG LTR sequences yielded the products of desired size. From these results it can be concluded that Genomic DNA of MCF-7 cell is not a suitable choice as positive control for PCR or RT-PCR based detection of MMTV-LS. It is also suggested that plasmids containing the cloned genes or sequences of MMTV be used as positive control for detection of MMTV-LS.
authors.affiliation: Department of Biotechnology and Microbiology, KLE's P C Jabin Science College, Hubli, Karnataka, India.
authors.forename: Bhushan B
authors.initials: BB
authors.lastname: Kulkarni
authors.forename: Shivaprakash V
authors.initials: SV
authors.lastname: Hiremath
authors.forename: Suyamindra S
authors.initials: SS
authors.lastname: Kulkarni
authors.forename: Umesh R
authors.initials: UR
authors.lastname: Hallikeri
authors.forename: Basavaraj R
authors.initials: BR
authors.lastname: Patil
authors.forename: Pramod B
authors.initials: PB
authors.lastname: Gai
mesh_terms: D001943:Breast Neoplasms / Q000175:diagnosis* / Q000821:virology; D045744:Cell Line, Tumor; D005260:Female; D006801:Humans; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics / Q000302:isolation & purification*; D057089:Pathology, Molecular / Q000379:methods* / Q000592:standards*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards*; D012015:Reference Standards; D014773:Virology / Q000379:methods / Q000592:standards
keywords: Breast; Cancer; MCF-7; MMTV; PCR; Positive control