Notch信号通路在骨髓间充质干细胞向肝细胞分化过程中的动态表达特征
柳柯, 刘桂英, 杨洋, 吴章林, 黄文华
中国临床解剖学杂志 ›› 2013, Vol. 31 ›› Issue (3) : 308-313.
Notch信号通路在骨髓间充质干细胞向肝细胞分化过程中的动态表达特征
Dynamic expression characteristics of Notch signal in bone marrow-derived mesenchymal stem cells during the process of differentiation into hepatocytes
目的 探寻Notch信号通路对BM-MSCs向肝细胞分化的影响机制。 方法 诱导BM-MSCs分化成肝细胞。当BM-MSCs分化至第0、7、11、21天时，反向斑点杂交实验检测Notch信号通路中的关键基因的mRNA表达水平。建立加入Jagged1上调信号通路的对照组RT-PCR技术绘制出BM-MSCs分化状态分子表达谱与正常情况下对比。 结果 BM-MSCs分化进行至第21天，反向斑点杂交检测到的关键基因的mRNA表达水平低于第0、7、11天。加入Jagged1后Notch信号通路被激活，导致下游基因Hes1 和 Hey1的被表达。BM-MSCs分化过程中Albumin未被检测到。 结论 本研究的结果表明Notch信号通路在BMMSCs分化成肝细胞过程进行调控是必需的，但是分化必须在信号通路下调的情况下才能进行下去。
Objective To explore the effect of Notch signaling in bone marrow-derived mesenchymal stem cells during the process of differentiation into hepatocytes. Methods An induction system under which BM-MSCs can differentiate into hepatocytes was established. On days 0, 7, 11 and 21 day during the differentiation direction, nine key genes in Notch pathway were selected as target genes and determined in BM-MSCs using technique reverse dot blot hybridization. The results of reverse dot blot hybridization showed that the effect of Notch signaling in BM-MSCs during the process of differentiation into hepatocytes. At the same time，the results of RT-PCR analysis reveals the differentiation status of BM-MSCs. Results On the 21th day when the differentiation direction was determined in BM-MSCs, the mRNA level of nine key genes was significantly lower than that on days 0, 7, and 11. In the further experiments, down-regulation of Notch signaling was shown to be critical for BM-MSCs to differentiate into hepatocytes, as increased Jagged1 resulted in up-regulated Notch activation, leading to higher levels of expression of Hes1 and Hey1, which completely blocked Albumin expresion in BM-MSCs. Conclusion These results in our study showed that Notch signaling in BM-MSCs was necessary to initiate differentiation into hepatocytes, but must be down-regulated for the differentiation to proceed continuously.
成体干细胞 / 间充在干细胞 / 定向分化 / 肝细胞 / Notch信号通路
Adult stem cell / Mesenchymal stem cell / Directing-differentiation / Hepatocyte / Notch
[1] Kashofer K, Bonnet D. Gene therapy progress and prospects: stem cell plasticity
[J]. Gene Ther，2005,12(16):1229-1234.
[2] Krause DS, Theise ND, Collector MI, et al. Multi-organ, multi-lineage engraftment by a single bone marrow-derived stem cell
[J]. Cell, 2001,105(7):369-377.
[3]Lee KD, Kuo TK, Lin T, et al. In vitro hepatic differentiation of human mesenchymal stemcel
[J]. Hepatology, 2004,40(3):1275-1284.
[4] Petersen BE, Bowen WC, Patrene KD, et al. Bone marrow as a potential source of hepatic oval cells
[J].Science,1999, 284(2):1168-1170.
[5] Hong H, Chen JZ, Zhou F, et al. Influence of serum from liver-damaged rats on differentiation tendency of bone marrow-derived stem cells
[J]. World J Gastroenterol, 2004, 10(6):2250-2253.
[6] Yuan TM, Yu HM. Notch signaling: key role in intrauterine infection/inflammation, embryonic development and white matter damage
[J]. Neurosci Res, 2010,88(3):461-468.
[7] Weinmaster G. The ins and outs of notch signaling
[J].Mol Cell Neurosci,1997, 9(5):91-102.
[8] Ward EJ, Shcherbata HR, Reynolds SH, et al. Stem cells signal to the niche through the Notch pathway in the Drosophila ovary
[J]. Curr Biol, 2006,16(6):2352-2358.
[9] Reddy BV, Rauskolb C, Irvine KD, et al. Influence of fat-hippo and notch signaling on the proliferationand differentiation of Drosophila optic neuroepithelia
[J]. Development, 2010,137(8):2397-2408.
[10] Wahl MB, Deng C, Lewandoski M, et al. FGF signaling acts upstream of the NOTCH and WNT signaling pathways to control segmentation clock oscillations in mouse somitogenesis
[J]. Development, 2007,134(3):4033-4041.
[11] Cau E, Blader P. Notch activity in the nervous system: to switch or not switch
[J]. Neural, 2009, 4(9):36.
[12] Sandy AR, Maillard I. Notch signaling in the hematopoietic system
[J]. Expert Opin Biol Ther, 2009, 9(5):1383-1398.
[13] Hong SH, Gang EJ, Jeong JA, et al. In vitro differentiation of human umbilical corb blood-derived mesenchymal stem cells intohepatocyte-like cells
[J]. Biochem Biophys Res Commun ,2005,330(8):1153-1161.
[14] Nwabo Kamdje AH, Mosna F, Bifari F, et al. Notch-3 and Notch-4 signaling rescue from apoptosis human B-ALL cells in contact with human bone marrow-derived mesenchymal stromal cells
[J]. Blood, 2011, 118(3): 380-389.
[15] Oyagi S, Hirose M, Kojima M, et al. Therapeutic effect of transplanting HGF-treated bone marrow mesenchymal cells into CCl4-injured rats
[J]. J Hepatol, 2006, 44(7):742-748.
[16] Oh SH, Witek RP, Bae SH, et al. Petersen BE.Bonemarrow- derivedhepatic oval cells differentiate into hepatocytes in 2-acetylaminofluorene/partial hepatectomy-induced liverregeneration
[J]. Gastroenterology, 2007,132(9):1077-1087.
[17] Podestà M, Benvenuto F, Pitto A, et al. Concentrative uptake of cyclic ADP-ribose generated by BST-1+ stroma stimulates proliferation of human hematopoietic progenitors
[J]. Biol Chem, 2005, 280(7):5343-5349.
[18] Weinhouse S. A single GST-P positive hepatocyte induced by a hepatocarcinogen
[J].Jpn J Cancer Res, 1991, 82(5):1464-1465.
[19] Lee SJ, Friedman SL, Whalen R, et al. Cellular sources of glutathione S-transferase P in primary cultured rat hepatocytes: localization by in situ hybridization
[J].Biochem J,1994, 299 (1):79-83.
[20] Tee LB, Kirilak Y, Huang WH, et al. Dual phenotypic expression of hepatocytes and bile ductular markers in developing and preneoplastic rat liver
[J].Carcinogenesis，1996,17(4):251-259.
[21]Kageyama R, Ishibashi M, Takebayashi K, et al. bHLH transcription factors and mammalian neuronal differentiation
[J] . Biochem Cell Biol,1997, 29(7):1389-1399.
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