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Archive > Volume 25 Issue 6 > 2018,25(6):607-612. DOI:10.3872/j.issn.1007-385X.2018.06.010 Prev Next
Basic Research
Establishment of cisplatin-resistant breast cancer cell line and role of FANCF gene in cisplatin resistance
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Abstract:
[Abstract] Objective: To investigate the expression profile and function of FANCF gene (a key gene in FA/BRCA pathway) in both cisplatin (DDP)-resistant and DDP-sensitive human triple-negative breast cancer cell lines and to analyze its correlation with DDP-resistance in breast cancer. Methods: The DDP-resistant breast cancer MDA-MB-231 cell line (MDA-MB-231/DDP) was established by induction of gradient DDP. The expression of FANCF gene in both sensitive and resistant cell lines was knocked-down by RNAi interference technology and the knockdown efficiency was validated at both RNA and protein level. The cell viability of MDA-MB-231 cells and MDA-MB-231/DDP cells was determined by the CCK8 assay; Flow cytometry was used to examine the cell cycle distribution and apoptosis; the mRNA and protein expressions of FANCF gene were examined by using qRT-PCR and western blotting, respectively. Results:The resistance index of MDA-MB-231 /DDP cells was 13.5 after 3-month induction. The mRNA and protein expressions of FANCF were significantly increased in MDA-MB-231/DDP cells (all P<0.01). Cell cycle analysis indicated that the DDP treatment significantly induced G0/G1 arrest and decreased the cell proportion in phase S and G2/M. siRNA-mediated knockdown of FANCF could not only be able to increase sensitivity of MDA-MB-231 to DDP but also promote the cell apoptosis (all P<0.01). Conclusion: FANCF attributes to the occurrence of DDP-resistance through anti-apoptosis effect, which might be served as a potential treatment target for drug-resistant human breast cancer.
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Project Supported:
Project supported by National Natural Science Foundation of China（No.81660472）, Health System Leading Personnel Training Program of Yunnan Province (No.L-201212)，and the Key Laboratory of Transformation Medicine of Cell Therapy Technology in Yunnan（No.2015DG034）
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