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Expression and stability of c-sis mRNA in human glioblastoma cells
R D Press1, D Samols, D A Goldthwait
1Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106.
Abstract:
The production of platelet-derived growth factor like (PDGF-like) material by glioblastomas may be involved in the conversion of normal cells to tumor cells. In an investigation of this problem, we have examined some of the properties of the platelet-derived growth factor B-chain mRNA (c-sis mRNA) by a sensitive and quantitative RNA-RNA solution hybridization method. In 5 out of 8 human glioblastoma cell lines, c-sis mRNA was present, and in the line with the highest level, there were approximately 4-10 molecules per cell. The half-lives of the c-sis mRNA in two glioblastoma cell lines were 2.6 and 3.4 h, while in human umbilical vein endothelial (HUVE) and bladder carcinoma (T24) cells they were 1.6 and 2.5 h, respectively. Inhibiting protein synthesis produced no significant alteration of the c-sis mRNA half-lives in the glioblastoma or HUVE cells. The A-U-rich sequence at the 3' end of the c-sis mRNA therefore does not appear to affect the mRNA stability in the presence of cycloheximide as it does in other transcripts. The similarity of the c-sis mRNA half-lives in normal and tumor cells suggests that regulation of stability of c-sis mRNA is not a major factor in tumorigenesis in the glioblastoma cell lines examined.
Insights
Glioblastoma cells produce platelet-derived growth factor B-chain mRNA (c-sis mRNA). Its stability in tumor cells is similar to normal cells, suggesting mRNA stability is not a primary driver of glioblastoma development.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Glioblastomas may transform normal cells into tumor cells by producing platelet-derived growth factor-like (PDGF-like) material.
- Platelet-derived growth factor B-chain mRNA (c-sis mRNA) is a key component in PDGF production.
Purpose of the Study:
- To investigate the properties of c-sis mRNA in human glioblastoma cell lines.
- To determine if c-sis mRNA stability plays a role in glioblastoma tumorigenesis.
Main Methods:
- Utilized a sensitive and quantitative RNA-RNA solution hybridization method to examine c-sis mRNA.
- Measured c-sis mRNA half-lives in glioblastoma, human umbilical vein endothelial (HUVE), and bladder carcinoma (T24) cells.
- Assessed the impact of protein synthesis inhibition (cycloheximide) on mRNA stability.
Main Results:
- c-sis mRNA was detected in 5 out of 8 human glioblastoma cell lines, with levels up to 4-10 molecules per cell.
- c-sis mRNA half-lives in glioblastoma cells (2.6-3.4 h) were comparable to those in normal cells (HUVE: 1.6 h, T24: 2.5 h).
- Inhibition of protein synthesis did not significantly alter c-sis mRNA half-lives in either normal or tumor cells, indicating the 3' A-U-rich sequence does not affect stability under these conditions.
Conclusions:
- The stability of c-sis mRNA in glioblastoma cells is similar to that in normal cells.
- Regulation of c-sis mRNA stability is unlikely to be a major factor in the tumorigenesis of the examined glioblastoma cell lines.