Effect of LINE1-ORF1p overexpression on the proliferation of nephroblastoma WT_CLS1 cells / 中国当代儿科杂志
Chinese Journal of Contemporary Pediatrics
;
(12): 501-507, 2018.
Article
in Chinese
| WPRIM
| ID: wpr-689599
ABSTRACT
<p><b>OBJECTIVE</b>To prepare the LINE1-ORF1p polyclonal antibody, and to study the effect of LINE1-ORF1p on the proliferation of nephroblastoma WT_CLS1 cells.</p><p><b>METHODS</b>A genetic engineering method was used to achieve prokaryotic expression of LINE1-ORF1p, and rabbits were immunized with LINE1-ORF1p to prepare polyclonal antibody. Indirect ELISA was used to evaluate antibody titer, and Western blot and immunohistochemistry were used to evaluate the specific ability of antibody to recognize LINE1-ORF1p. The eukaryotic expression vector pEGFP-N1-LINE1-ORF1 was constructed and used to transfect WT_CLS1 cells. Western blot and qRT-PCR were used to measure the protein and mRNA expression of LINE1-ORF1, respectively, and cell proliferation assay and colony-forming assay were used to evaluate the effect of LINE1-ORF1p on the proliferation of WT_CLS1 cells and the formation of tumor cell clone.</p><p><b>RESULTS</b>The LINE1-ORF1p antibody prepared had a titer of >116 000 and could specifically recognize LINE1-ORF1p in cells and tumor tissue. WT_CLS1 cells transfected with pEGFP-N1-LINE1-ORF1 had significant increases in the mRNA and protein expression of LINE1-ORF1 and significantly enhanced cell proliferation ability and colony formation ability (P<0.05).</p><p><b>CONCLUSIONS</b>LINE1-ORF1p can promote the growth of nephroblastoma cells and the formation of tumor cell clone, and may be involved in the pathogenesis of nephroblastoma.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
RNA, Messenger
/
Transfection
/
Blotting, Western
/
Wilms Tumor
/
Long Interspersed Nucleotide Elements
/
Cell Line, Tumor
/
Deoxyribonuclease I
/
Cell Proliferation
/
Genetics
/
Metabolism
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Contemporary Pediatrics
Year:
2018
Type:
Article
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