| Objective: To investigate the expression and mutation of tumor susceptibilitygenelOl(TSGlOl) and its 4 splice variants (V_A,V_B,V_C and V_D) in peripheral blood cell of healthy women in order to explore the role of TSGlOl in tumorigenesis and development of breast cancer.Methods: Total RNA was isolated from peripheral blood mononuclear cells donatedby 37 healthy women. Full length of gene TSGlOl and its splice variants (V_A,V_B, V_C and V_D) were detected by RT-PCR using 5 pairs of specific primers. Agarose electrophoresis was performed to identify the PCR products. Each products was sequenced with primers ( covered 663bp-1276bp) and compared with the sequence of gene TSGlOl issued in genebank; PCR amplification of genomic DNA which was extracted from some blood samples was performed, using specific primers for exon 4 of gene TSG101 and PCR products were also sequenced.Results: Full length and 4 splice variants of gene TSGlOl had been detected byRT-PCR in all of 37 healthy women blood mononuclear cells . Among them, the different expression was shown, with the most prominent difference in full length and, while lesser in V_D of gene TSG101,and least in V_A, V_B and V_C; Sequence results showed that the sequence of 30 samples (30/37,81.08%) were 100% matching the sequence in genebank, while in other 7 samples (7/37,18.92%) a same "C→T" heterozygous point mutation, at 798bp of exon 4, was detected. Meanwhile, at genomic DNA level, the same mutation in the individual with point mutation in RNA was confirmed.Conclusions: l.Full length of gene TSG101 and its four splice variants (VA% VB>Vcand Vd) were detected in blood mononuclear cells of all 37 healthy women. 2. A "C—>T" heterozygous point mutation locating at 798bp of mRNA in healthy women, the present data might suggest that single nucleotide polymorphisms exist in gene TSG101. The relationship between heterozygous point mutation and breast cancer need further studying.
|
PDF Full Text Request