Increase the cisplatin cytotoxicity and cisplatin-induced DNA damage in HepG2 cells by XRCC1 abrogation related mechanisms.
Cisplatin is one of the most potent chemotherapeutic anticancer drugs for the treatment of various cancers. The cytotoxic action of the drug is often thought to be associated with its ability to bind DNA to form cisplatin-DNA adducts. Impaired DNA repair processes including base excision repair (BER) play important roles on its cytotoxicity. XRCC1 is a key protein known to play a central role at an early stage in the BER pathway. However, whether XRCC1 contributes to decrease the cisplatin cytotoxicity and cisplatin-induced DNA damage in HepG2 still remains unknown. Hence, the purpose of this study was to explore whether abrogation of XRCC1 gene expression by short hairpin RNAs (shRNA) could reduce DNA repair and thus sensitize liver cancer cells to cisplatin. We abrogated the XRCC1 gene in HepG2 cell using shRNA transfection. Cell viability was measured by MTT assay and clonogenicity assay. Comet assay was used to detect the DNA damage induced by cisplatin. The host cell reactivation was employed to assess the DNA repair capacity of cisplatin-damaged luciferase reporter plasmid. Flow cytometry analysis was used to determine cisplatin-induced apoptosis, cell cycle and reactive oxygen species (ROS). The results showed that abrogation of XRCC1 could sensitize HepG2 cells to cisplatin. This enhanced cytotoxicity could be attributed to the increased DNA damage and reduced DNA repair capacity. Increasing cell cycle arrest and intracellular ROS production lead to more tumor cell apoptosis and then enhanced the cisplatin cytotoxicity. Our results suggested that the cisplatin cytotoxicity may increase by targeting inhibition of XRCC1.
Department of Occupational Health and Environmental Health, School of Public Health, Hebei Medical University, Shijiazhuang, Hebei, People's Republic of China. rong_zhang1@yahoo.com.cn
Rong
R
Zhang
Yujie
Y
Niu
Yikai
Y
Zhou
D000970:Antineoplastic Agents / Q000633:toxicity*; D002453:Cell Cycle / Q000187:drug effects; D002945:Cisplatin / Q000633:toxicity*; D003603:Cytotoxins / Q000633:toxicity*; D018736:DNA Adducts / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D056945:Hep G2 Cells; D006801:Humans; D018384:Oxidative Stress / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D000076105:X-ray Repair Cross Complementing Protein 1
D000970:Antineoplastic Agents; D003603:Cytotoxins; D018736:DNA Adducts; D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D000076105:X-ray Repair Cross Complementing Protein 1; C000619371:XRCC1 protein, human; D002945:Cisplatin