Objective： To investigate the changes of cyclin L2（CCNL2） gene mRNA and protein during the differentiation of P19 cells to cardiac myocytes, and to explore the relationship between CCNL2 gene and the differentiatio...Objective： To investigate the changes of cyclin L2（CCNL2） gene mRNA and protein during the differentiation of P19 cells to cardiac myocytes, and to explore the relationship between CCNL2 gene and the differentiation of cardiac myocytes. Methods： P19 cells were cultured with 0.9% DMSO for 18 days. Western blots of cardiac troponin I （cTnI） were used to identify cell differentiation. Total RNA was extracted from P19 cells during the process of differentiation at various time points：pre-differentiation（Day 0）, and Day 1 to Day 18. The expression levels of CCNL2 gene mRNA and protein were evaluated by RT-PCR and Western blot, respectively. Results： After being induced to differentiate by DMSO for 4 days in suspension, spontaneously and rhythmically beating ceils were seen at 8 day, which were cTnI-positive. In P19 cells, both the expression level of CCNL2 gene mRNA and protein were gradually down-regulated. Conclusion： Both the expression of CCNL2 gene and protein were down-regulated during the process of the differentiation of P19 cells into cardiac myocytes, suggesting a possible role for this cyclin in their differentiation.展开更多
Maintenance of pluripotency depends to diverse regulatory factors.Studies in embryonic stem cells(ESCs)have indicated that large intergenic non-coding RNAs(lincRNAs)are involved in the regulatory network of pluripoten...Maintenance of pluripotency depends to diverse regulatory factors.Studies in embryonic stem cells(ESCs)have indicated that large intergenic non-coding RNAs(lincRNAs)are involved in the regulatory network of pluripotency.However,the presence and function of pluripotency-associated lincRNAs in cancer cells with pluripotency features are unknown.In this study,we used embryonal carcinoma(EC)P19 cell lines to investigate the expression level of Halr1 in pluripotency and retinoic acid(RA)-induced differentiated states.Down-regulation of pluripotency associated factors such as OCT4,NANOG,SSEA1 and alkaline phosphatase at transcript and protein levels were used to confirm the differentiated status of P19 cells.Quantitative measurement of Halr1 transcript levels revealed a 79% decrease during RA-induced differentiation of P19 cells.These results indicate that upon exiting the pluripotency state the expression level of Halr1 similar to core pluripotency factors is remarkably reduced.展开更多
文摘Objective： To investigate the changes of cyclin L2（CCNL2） gene mRNA and protein during the differentiation of P19 cells to cardiac myocytes, and to explore the relationship between CCNL2 gene and the differentiation of cardiac myocytes. Methods： P19 cells were cultured with 0.9% DMSO for 18 days. Western blots of cardiac troponin I （cTnI） were used to identify cell differentiation. Total RNA was extracted from P19 cells during the process of differentiation at various time points：pre-differentiation（Day 0）, and Day 1 to Day 18. The expression levels of CCNL2 gene mRNA and protein were evaluated by RT-PCR and Western blot, respectively. Results： After being induced to differentiate by DMSO for 4 days in suspension, spontaneously and rhythmically beating ceils were seen at 8 day, which were cTnI-positive. In P19 cells, both the expression level of CCNL2 gene mRNA and protein were gradually down-regulated. Conclusion： Both the expression of CCNL2 gene and protein were down-regulated during the process of the differentiation of P19 cells into cardiac myocytes, suggesting a possible role for this cyclin in their differentiation.
基金supported by a Ferdowsi University of Mashhad grant to HD(No.22534).
文摘Maintenance of pluripotency depends to diverse regulatory factors.Studies in embryonic stem cells(ESCs)have indicated that large intergenic non-coding RNAs(lincRNAs)are involved in the regulatory network of pluripotency.However,the presence and function of pluripotency-associated lincRNAs in cancer cells with pluripotency features are unknown.In this study,we used embryonal carcinoma(EC)P19 cell lines to investigate the expression level of Halr1 in pluripotency and retinoic acid(RA)-induced differentiated states.Down-regulation of pluripotency associated factors such as OCT4,NANOG,SSEA1 and alkaline phosphatase at transcript and protein levels were used to confirm the differentiated status of P19 cells.Quantitative measurement of Halr1 transcript levels revealed a 79% decrease during RA-induced differentiation of P19 cells.These results indicate that upon exiting the pluripotency state the expression level of Halr1 similar to core pluripotency factors is remarkably reduced.
