Abstract:
:DNA damage sensor proteins are recognized as upstream components of the DNA damage checkpoint signaling pathway and are required for cell cycle control and the induction of apoptosis. hRad9 plays an important role as an upstream regulator of checkpoint signaling. In our previous studies, we confirmed the significant accumulation of hRad9 in the nuclei of tumor cells in surgically-resected non-small cell lung cancer (NSCLC) specimens. We also found that the capacity to produce a functional hRad9 protein was intact in lung cancer cells, a finding which suggests that hRad9 would be a vital component in the pathways that lead to the survival and progression of NSCLC. Small interfering RNA targeting hRad9 was transfected into human lung adenocarcinoma A549 and PC3 cells. After the hRad9 knockdown, the cytotoxicity of the transfected cells was measured by a neutral red uptake test, and the G2-M arrest of irradiated cells was examined by flow cytometry. Significant cytotoxicity was observed in the cancer cells in which hRad9 expression was down-regulated. We also detected the inhibition of Chk1 phosphorylation by Western blot analysis. This suggested that hRad9 silencing leads to the impairment of the DNA damage checkpoint signaling pathway in tumor cells. Flow cytometry indicated a reduced population of cells in the G2-M phase, an observation consistent with the findings of several studies that indicated that hRad9 is necessary for G2-M arrest. In conclusion, the current study demonstrated that RNA interference targeting hRad9 in cancer cells leads to the impairment of the DNA damage checkpoint signaling pathway, which appears to be essential for maintaining tumor cell proliferation, and induces cell death. Therefore, hRad9 may be a novel molecular target for lung cancer treatment.
journal_name
Oncol Repjournal_title
Oncology reportsauthors
Yuki T,Maniwa Y,Doi T,Okada K,Nishio W,Hayashi Y,Okita Ysubject
Has Abstractpub_date
2008-11-01 00:00:00pages
1047-52issue
5eissn
1021-335Xissn
1791-2431journal_volume
20pub_type
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