pSUPER-siRNA-IGF1R protein enhances sensitivity of human liver cancer cells to mitomycin in vitro
- Author:
Jian NIU
1
Author Information
1. Department of General Surgery
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Liver neoplasms, experimental;
Receptors,insulin-like growth factors;
RNA, small molecular interfering
- From:
Tumor
2007;27(11):870-873
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate whether the plasmid of short hairpin RNA targeting human insulin like growth factor receptor 1 (pSUPER-siRNA-IGF1R) can enhance mitomycin-induced apoptosis of human liver cancer SMMC7721 cells and the corresponding mechanism. Methods: The siRNA targeting IGF1R was designed and the pSUPER-siRNA-IGFI R plasmid was constructed and transfected into SMMC7721 cells. The stable cell lines were screened by G418. The growth curve of every group was detected by CCK8 method. After mitomycin treatment, the sensitivity of hepatoma cells to chemotherapy was monitored by CCK8 method. The apoptotic index (AI) was examined by flow cytometry. The expression of apoptosis-related proteins p53, Bax, bcl-2 was determined by Western blotting. The empty control group and positive control were set up. Results: The apoptosis induced by mitomycin were significantly enhanced in stable cell line and the AI was significantly elevated compared with other groups (P < 0.05). The expression of wild type p53 and Bax protein was significantly up-regulated but the expression of bcl-2 protein was significantly down-regulated (P < 0.05). Conclusion: The constructed pSUPER-siRNA-IGF1R plasmid silences RNA transcription and enhances the apoptosis of tumor cells induced by mitomycin.
