scholarly journals RADIOBIOLOGIC EVALUATION OF AN IN VITRO MAMMALIAN CELL RENEWAL SYSTEM. Terminal Progress Report.

1972 ◽  
Author(s):  
G. V. Dalrymple ◽  
J. L. Sanders ◽  
J. C. Nash
Keyword(s):  
Mammalian Cell ◽  
Progress Report ◽  
Cell Renewal ◽  
Renewal System

A novel PLK1 inhibitor onvansertib effectively sensitizes MYC-driven medulloblastoma to radiotherapy

2021 ◽  
Author(s):  
Dong Wang ◽  
Bethany Veo ◽  
Angela Pierce ◽  
Susan Fosmire ◽  
Krishna Madhavan ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Dna Damage ◽  
Tumor Growth ◽  
Radiation Treatment ◽  
Tumor Regression ◽  
Tumor Growth Inhibition ◽  
Cell Cycle Distribution ◽  
Cell Renewal ◽  
Group 3

Abstract Background Group 3 medulloblastoma (MB) is often accompanied by MYC amplification. PLK1 is an oncogenic kinase that controls cell cycle and proliferation and has been preclinically validated as a cancer therapeutic target. Onvansertib (PCM-075) is a novel, orally available PLK1 inhibitor, which shows tumor growth inhibition in various types of cancer. We aim to explore the effect of onvansertib on MYC-driven medulloblastoma as a monotherapy or in combination with radiation. Methods Crisper-Cas9 screen was used to discover essential genes for MB tumor growth. Microarray and immunohistochemistry on pediatric patient samples were performed to examine the expression of PLK1. The effect of onvansertib in vitro was measure by cell viability, colony-forming assays, extreme limiting dilution assay and RNA-Seq. ALDH activity, cell-cycle distribution and apoptosis were analyzed by flow cytometry. DNA damage was assessed by immunofluorescence staining. Medulloblastoma xenografts were generated to explore the monotherapy or radio-sensitizing effect. Results PLK1 is overexpressed in Group 3 MB. The IC50 concentrations of onvansertib in Group 3 MB cell lines were in a low nanomolar range. Onvansertib reduced colony formation, cell proliferation, stem cell renewal and induced G2/M arrest in vitro. Moreover, onvansertib in combination with radiation increased DNA damage and apoptosis compare with radiation treatment alone. The combination radiotherapy resulted in marked tumor regression in xenografts. Conclusions These findings demonstrate the efficacy of a novel PLK1 inhibitor onvansertib in vitro and in xenografts of Group 3 MB, which suggests onvansertib is an effective strategy as monotherapy or in combination with radiotherapy in MB.

In Vitro Base Excision Repair Assay Using Mammalian Cell Extracts

1999 ◽  
pp. 301-315 ◽  
Author(s):  
Guido Frosina ◽  
Enrico Cappelli ◽  
Paola Fortini ◽  
Eugenia Dogliotti
Keyword(s):  
Mammalian Cell ◽  
Base Excision Repair ◽  
Excision Repair ◽  
Cell Extracts ◽  
Base Excision

Efficient long DNA gap-filling in a mammalian cell-free system: A potential new in vitro DNA replication assay

Biochimie ◽  
2013 ◽  
Vol 95 (2) ◽  
pp. 320-328
Author(s):  
Seiki Nakao ◽  
Sufang Zhang ◽  
Markku Vaara ◽  
Juhani E. Syväoja ◽  
Marietta Y. Lee ◽  
...  
Keyword(s):  
Dna Replication ◽  
Mammalian Cell ◽  
Gap Filling ◽  
Free System ◽  
Cell Free System ◽  
Replication Assay ◽  
System A

scholarly journals Generation of single-nucleotide repair patches following excision of uracil residues from DNA.

1992 ◽  
Vol 12 (4) ◽  
pp. 1605-1612 ◽  
Author(s):  
G Dianov ◽  
A Price ◽  
T Lindahl
Keyword(s):  
Mammalian Cell ◽  
Excision Repair ◽  
Enzymatic Digestion ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Repair Synthesis ◽  
Single Nucleotide ◽  
Cell Extracts ◽  
Dna Repair Synthesis

The extent and location of DNA repair synthesis in a double-stranded oligonucleotide containing a single dUMP residue have been determined. Gently prepared Escherichia coli and mammalian cell extracts were employed for excision repair in vitro. The size of the resynthesized patch was estimated by restriction enzyme analysis of the repaired oligonucleotide. Following enzymatic digestion and denaturing gel electrophoresis, the extent of incorporation of radioactively labeled nucleotides in the vicinity of the lesion was determined by autoradiography. Cell extracts of E. coli and of human cell lines were shown to carry out repair mainly by replacing a single nucleotide. No significant repair replication on the 5' side of the lesion was observed. The data indicate that, after cleavage of the dUMP residue by uracil-DNA glycosylase and incision of the resultant apurinic-apyrimidinic site by an apurinic-apyrimidinic endonuclease activity, the excision step is catalyzed usually by a DNA deoxyribophosphodiesterase rather than by an exonuclease. Gap-filling and ligation complete the repair reaction. Experiments with enzyme inhibitors in mammalian cell extracts suggest that the repair replication step is catalyzed by DNA polymerase beta.

scholarly journals Translation of Reovirus Messenger RNAs Synthesized In Vitro into Reovirus Polypeptides by Several Mammalian Cell-Free Extracts

1972 ◽  
Vol 69 (9) ◽  
pp. 2649-2653 ◽  
Author(s):  
M. J. McDowell ◽  
W. K. Joklik ◽  
L. Villa-Komaroff ◽  
H. F. Lodish
Keyword(s):  
Mammalian Cell ◽  
Messenger Rnas

Messenger activity in mammalian cell-free extracts of reovirus single-stranded RNA prepared in vitro

1971 ◽  
Vol 42 (3) ◽  
pp. 454-461 ◽  
Author(s):  
Daniel H. Levin ◽  
David Kyner ◽  
George Acs ◽  
Samuel Silverstein
Keyword(s):  
Mammalian Cell
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