[Study on the expression of DNA excision repair biomarkers in cispatin-treated lung cancer cell line].
OBJECTIVE
To study the expression levels of ERCC2, UDG, and PCNA in cisplatin-treated A549 cell line.


METHODS
Comet assay, RT-PCR, and western blot were used to study the mRNA and protein expression levels of ERCC2, UDG, and PCNA.


RESULTS
When treated with IC(20) cisplatin, the DNA damage level increased as the cisplatin treated time increased within 24 h of cisplatin treatment. The tail state 12 h and 24 h after treatment was 5.02 +/- 0.68 and 7.22 +/- 0.53 respectively, which was significantly higher than those of the controls (2.73 +/- 0.29). The tail state 24 h after treatment was not significantly different from that of the controls. The DNA damage level decreased to normal after cisplatin treatment in 24 h (tail state 3.64 +/- 0.7). The expression levels of ERCC2, UDG, PCNA protein (4.37 +/- 0.57, 5.47 +/- 0.46, 2.21 +/- 0.47 respectively) and mRNA (0.71 +/- 0.08, 0.74 +/- 0.06, 0.82 +/- 0.09) were increased significantly within 24 h exposure and decreased to normal 24 h after cisplatin treatment. The 3 enzymes' mRNA and protein expression increased when treated with cisplatin, but the changes of protein level were slower than those of mRNA levels.


CONCLUSIONS
The DNA repair capability in A549 cells increases after cisplatin treatment. Cisplatin was a positive regulation of ERCC2, UDG, PCNA expression levels, which causes the increase of mRNA, and protein. The positive regulation only works in a short time and returns normal after 24 h of cisplatin treatment.
Wang
Zhi-yong
ZY
Institute of Environmental medicine, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
Zhou
Sheng
S
Lu
Bin
B
Zhang
Xia
X
Hao
Qiao-ling
QL
Zhou
Yi-kai
YK
D000970:Antineoplastic Agents / Q000494:pharmacology*; D014408:Biomarkers, Tumor; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D020552:Comet Assay; D045647:DNA Glycosylases / Q000096:biosynthesis / Q000235:genetics; D004265:DNA Helicases; D004260:DNA Repair / Q000187:drug effects*; D004268:DNA-Binding Proteins; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D018809:Proliferating Cell Nuclear Antigen / Q000096:biosynthesis / Q000235:genetics; D014176:Protein Biosynthesis; D011506:Proteins / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis; D014157:Transcription Factors; D051759:Xeroderma Pigmentosum Group D Protein
D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D004268:DNA-Binding Proteins; D018809:Proliferating Cell Nuclear Antigen; D011506:Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D045647:DNA Glycosylases; D004265:DNA Helicases; D051759:Xeroderma Pigmentosum Group D Protein; C497292:ERCC2 protein, human; D002945:Cisplatin