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Complementation of the radiosensitive M059J cell line.

B S Hoppe¹, R B Jensen, C U Kirchgessner

¹Mayer Cancer Research Laboratory, Department of Radiation Oncology, Stanford University School of Medicine, Stanford, California 94305, USA.

Radiation Research

|January 12, 2000

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The radiosensitive M059J glioblastoma cell line has a defect in the DNA-dependent protein kinase catalytic subunit (PRKDC). Complementation with the PRKDC gene restored kinase activity and radioresistance, confirming this defect.

Area of Science:

Molecular Biology
Cancer Research
Cell Biology

Background:

M059J and M059K are glioblastoma cell lines derived from the same tumor.
M059J is radiosensitive, while M059K is radioresistant.
M059J cells lack expression of the DNA-dependent protein kinase catalytic subunit (PRKDC).

Purpose of the Study:

To investigate the cause of radiosensitivity in M059J cells.
To determine if PRKDC deficiency underlies the radiosensitive phenotype.
To establish complemented cell lines for further DNA-PK studies.

Main Methods:

Complementation of M059J cells with a human chromosome 8 fragment containing the PRKDC gene.
Generation of hybrid cell lines retaining the introduced PRKDC gene.

Related Experiment Videos

Assay of kinase activity and radiosensitivity in hybrid cell lines.

Main Results:

Hybrid cell lines expressing PRKDC showed restored kinase activity.
These complemented cell lines exhibited radioresistance.
This demonstrates that PRKDC deficiency is the cause of M059J radiosensitivity.

Conclusions:

The primary defect in M059J cells is the lack of functional PRKDC.
Complemented M059J cell lines are valuable tools for studying DNA-PK function.
These cell lines offer a closer genetic match to M059J for research compared to M059K.