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12 June 2016 : Laboratory Research

DNA Repair Genes ERCC1 and BRCA1 Expression in Non-Small Cell Lung Cancer Chemotherapy Drug Resistance

Shuai WangBC, Feng LiuCD, Jingyan ZhuDE, Peng ChenEF, Hongxing LiuFG, Qi LiuBG, Junqing HanABCD

DOI: 10.12659/MSM.896606

Med Sci Monit 2016; 22:1999-2005

Abstract

BACKGROUND: Surgery combined with chemotherapy is an important therapy for non-small cell lung cancer (NSCLC). However, chemotherapy drug resistance seriously hinders the curative effect. Studies show that DNA repair genes ERCC1 and BRCA1 are associated with NSCLC chemotherapy, but their expression and mechanism in NSCLC chemotherapy drug-resistant cells has not been elucidated.

MATERIAL AND METHODS: NSCLC cell line A549 and drug resistance cell line A549/DDP were cultured. Real-time PCR and Western blot analyses were used to detect ERCC1 and BRCA1 mRNA expression. A549/DDP cells were randomly divided into 3 groups: the control group; the siRNA-negative control group (scramble group); and the siRNA ERCC1 and BRCA1siRNA transfection group. Real-time PCR and Western blot analyses were used to determine ERCC1 and BRCA1 mRNA and protein expression. MTT was used to detect cell proliferation activity. Caspase 3 activity was tested by use of a kit. Western blot analysis was performed to detect PI3K, AKT, phosphorylated PI3K, and phosphorylated AKT protein expression.

RESULTS: ERCC1 and BRCA1 were overexpressed in A549/DDP compared with A549 (P<0.05). ERCC1 and BRCA1siRNA transfection can significantly reduce ERCC1 and BRCA1 mRNA and protein expression (P<0.05). Downregulating ERCC1 and BRCA1 expression obviously inhibited cell proliferation and increased caspase 3 activity (P<0.05). Downregulating ERCC1 and BRCA1 significantly decreased PI3K and AKT phosphorylation levels (P<0.05).

CONCLUSIONS: ERCC1 and BRCA1 were overexpressed in NSCLC drug-resistant cells, and they regulated lung cancer occurrence and development through the phosphorylating PI3K/AKT signaling pathway.

Keywords: Antineoplastic Combined Chemotherapy Protocols - pharmacology, A549 cells, BRCA1 Protein - genetics, Carcinoma, Non-Small-Cell Lung - genetics, DNA Repair - genetics, DNA-Binding Proteins - genetics, Down-Regulation - drug effects, Drug Resistance, Neoplasm - genetics, Endonucleases - genetics, Lung Neoplasms - genetics, RNA, Messenger - genetics, RNA, Small Interfering - genetics, Real-Time Polymerase Chain Reaction

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750