ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect and regulatory mechanism of clock gene Per1 on the proliferation,apoptosis,migration,and invasion of human oral squamous carcinoma SCC15 cells.</p><p><b>METHODS</b>RNA interference was used to knock down Per1 gene in human oral squamous cell carcinoma SCC15 cell line. Changes of cell proliferation and apoptosis were analyzed by flow cytometry. Transwell assay was carried out to assess cell migration and invasion. Real-time polymerase chain reaction was used to detect the mRNA expressions of Ki-67, murine double minute 2 (MDM2), c-Myc, p53, Bax, Bcl-2, metalloproteinase (MMP)2, MMP9, and vascular endothelial growth factor (VEGF).</p><p><b>RESULTS</b>shRNA-mediated knockdown of Per1 promoted the proliferation, migration and invasion capacity, and inhibited cell apoptosis capacity of SCC15 cells (all P<0.05). Additionally, Per1 knockdown also increased the mRNA expressions of Ki-67, MDM2, Bcl-2, MMP2, and MMP9 and decreased the mRNA expressions of c-Myc, p53, and Bax (all P<0.05); however, the VEGF mRNA expression did not differ significantly after Per1 knockdown (P>0.05).</p><p><b>CONCLUSIONS</b>Clock gene Perl can regulate important tumor-related genes downstream such as Ki-67, MDM2, c-Myc, p53, Bax, Bcl-2, MMP2, and MMP9, and the aberrant expression of Per1 can affect tumor cell proliferation,apoptosis,migration and invasion. An in-depth study of Per1 may further clarify the mechanism of tumorigenesis and tumor development and thus provides new effective molecular targets for cancer treatment.</p>
Subject(s)
Humans , Apoptosis , Carcinoma, Squamous Cell , Metabolism , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Knockdown Techniques , Ki-67 Antigen , Metabolism , Matrix Metalloproteinase 2 , Metabolism , Matrix Metalloproteinase 9 , Metabolism , Period Circadian Proteins , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Proto-Oncogene Proteins c-mdm2 , Metabolism , Proto-Oncogene Proteins c-myc , Metabolism , RNA Interference , Real-Time Polymerase Chain Reaction , Tumor Suppressor Protein p53 , Metabolism , Vascular Endothelial Growth Factor A , Metabolism , bcl-2-Associated X Protein , MetabolismABSTRACT
Objective To explore the molecular evolution of myo-inositol oxygenase (Miox) gene and its temporal and spatial expression patterns during the development of Xenopus laevis embryos. Methods The temporal and spatial expression patterns of Miox gene were analyzed by semi, quantitative RT, PCR and whole, mount in situ hybridization technique, respectively. Results RT, PCR results showed that Miox gene was hardly found before stage 26; slight expression was found at stage 28, which gradually increased thereafter, reaching a high level at stage 40 and peaked at stage 41; and then it had a decrease at stage 45. Compared with stages 28, 34, stages 40, 41, and 45 had a significantly higher Miox gene expression (P<0.05). Compared with stage 40, stage 41 had a significantly higher Miox gene expression(P<0.05). But stage 45 had a significantly lower expression compared with stage 41(P<0.05). The results of whole, mount in situ hybridization showed no Miox expression before stage 30; at stage 33 weak expression was found in the pronephros, and the expression gradually increased as time went by. The results of whole, mount in situ hybridization were consistent with that of RT, PCR, with Miox expression notably increased at stage 39, 40, and then remained at that level. We also found that Miox was only expressed in the pronephros tubules during the whole embryo development period. Conclusion Miox is a kidney, specific gene during Xenopus laevis pronephros development, and it may serve as a marker for later pronephros development in organogenesis.