A preliminary study on ASPP1 mRNA expression level in tumor cell line
- VernacularTitle:ASPP1基因mRNA在肿瘤细胞株中的表达初探
- Author:
Bin SUN
;
Yun ZHANG
- Publication Type:Journal Article
- Keywords:
apoptosis stimulating protein of p53;
RNA,messager;
polymerase chain reaction;
tumor cells,cultured
- From:
Medical Journal of Chinese People's Liberation Army
2001;0(09):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a reverse transcription-PCR(RT-PCR) assay for detecting ASPP1 mRNA expression levels in normal blood monocyte and leukemia jurkat,HL-60 and K-562 cell lines.And to analyze the differences in relative expression levels of ASPP1 in respective cell line by calculating the value of ASPP1/?-actin for evaluating the effects of ASPP1 on inhibiting the malignant transformation of normal cells.Methods The monocyte isolation reagent was used to isolate normal blood monocyte,and the RPMI 1640 medium was used to culture jurkat,HL-60 and K562 cell lines.The cell RNA was extracted with TripureTM reagent and amplified by RT-PCR.The amplified products were E.B.stained and 10?l of the products were used for agarose gel electrophoresis.The expression levels of ASPP1 mRNA and ?-actin in respective cell line were measured and then compared.Result The ASPP1 mRNA was expressed both in normal blood monocyte and all the leukemic cells.The value of ASPP1/?-actin was 0.545?0.019 in normal blood monocyte,and it was 0.155?0.081,0.199?0.016 and 0.191?0.015,respectively,in the leukemic cells jurkat,HL-60,and K-562.The expression level of ASPP1 mRNA was higher in normal cells than that in the three leukemic cell lines.Conclusion On the process of tumor development,an ASPP1-p53 compound is formed by collaborated effects of ASPP1 and p53,which may act on the arch-apoptosis gene promoter,specifically enhance the ability of p53 binding DNA,and reinforce the function of p53 in enhancing apoptosis.The declined expression and/or deprived function of ASPP1 might lead to tumorigenesis,implying that the function of ASPP1 in enhancing p53 apoptosis may play an important role on inhibiting the malignant transformation of normal cells.
