Effect on retardation of G2/M phase in esophageal carcinoma cells transfected with CHK1 and CHK2 shRNA after irradiation

• Published in: Zhōnghuá zhŏngliú zázhì Vol. 28; no. 8; p. 572
• Main Authors: Wang, Yu-xiang, Zhu, Shu-chai, Feng, Wei, Li, Juan, Su, Jing-wei, Li, Ren
• Format: Journal Article
• Language: Chinese
• Published: China 01.08.2006
• Subjects: Blotting, Western; Cell Division - genetics; Cell Division - physiology; Cell Division - radiation effects; Cell Line, Tumor; Cell Survival - genetics; Cell Survival - physiology; Cell Survival - radiation effects; Checkpoint Kinase 1; Checkpoint Kinase 2; Esophageal Neoplasms - genetics; Esophageal Neoplasms - pathology; Esophageal Neoplasms - physiopathology; G2 Phase - genetics; G2 Phase - physiology; G2 Phase - radiation effects; Gamma Rays; Genetic Vectors; Humans; Protein Kinases - genetics; Protein Kinases - metabolism; Protein-Serine-Threonine Kinases - genetics; Protein-Serine-Threonine Kinases - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; RNA, Small Interfering - genetics; Transfection
• ISSN: 0253-3766

To observe the effect of RNA interference on CHK1 and CHK2 expression and change of G2/M phase arrest in esophageal carcinoma cells after irradiation. Four sequences short hairhip RNA (shRNA) of each CHK1 and CHK2 genes were constructed and connected with vector of pENTR/U6 plasmid, respectively, and then transfected into Eca109 cells with lipofectamine 2000 reagent. Protein and mRNA expression of CHK1 and CHK2 genes were detected with Western blotting and RT-PCR, respectively. Cell cycling was measured by flow cytometry after 5 Gy irradiation. Cell survival rate after 5 Gy irradiation was evaluated by clonegenetic assay. Four shRNA vector each of CHK1 and CHK2 genes were successfully constructed and transfected into Ecal09 cells, respectively. Protein expression of CHK1 and CHK2 were obviously decreased. Their mRNA expressions were also decreased after transfected with shRNA of CHK1 and CHK2. Arrest of G2/M stage in Eca109 cells were obviously decreased only in cells transfected with CHK1 shRNA but not with