Expression of P21Cip/WAF1, P27Kip1 and proliferating cell nuclear antigen during differentiation of human bone marrow mesenchymal stem cells into hepatocytes*,*,☆

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P21Cip/WAF1 and P27Kip1 are two important negative regulatory factors of cell cycle. Proliferating cell nuclear antigen (PCNA) is an index reflecting stem cell proliferation.


To study the expression of P21Cip/WAF1, P27Kip1and PCNA gene during differentiation of human bone marrow mensenchymal stem cells (BMSCs) into hepatocytes in vitro.


The cytology, gene level, observation experiment was conducted at the Laboratory of Linbaixin Medical Institute Center, Second Affiliated Hospital, Sun Yat-sen University from December 2006 to June 2007.


Bone marrow was collected from healthy donors undergoing bone marrow aspiration at the Second Affiliated Hospital, Sun Yat-sen University.


Human BMSCs were isolated by the whole bone marrow adherence method in vitro. At passage 5, BMSCs were incubated in opti-MEM I low-serum medium containing 20 μ g/L hepatocyte growth factor, 10 μ g/L fibroblast growth factor-4 to induce the differentiation into hepatocytes.


After induction, the expression of P21Cip/WAF1, P27Kip1and PCNA mRNA in BMSCs was detected by RT-PCR.


After BMSCs were induced for 0 day, 1 day, 3 days, 5 days, 7 days, the relative P21Cip/WAF1 mRNA value gradually increased (P < 0.01), but no significant differences in relative P27Kip1 mRNA value (P > 0.05). The relative PCNA mRNA value gradually decreased (P < 0.01), except no significant differences were detected at 0 and 1 day (P > 0.05).


Overexpression of P21Cip/WAF1 inhibits BMSC proliferation, and has a regulatory effect during the differentiation of BMSCs into hepatocytes. P27Kip1 may has no effects. PCNA expression has an opposite effect compared to P21Cip/WAF1, resulting in a gradually decreased tendency.

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