1,25(OH)2 vitamin D3 induces elevated expression of the cell cycle-regulating genes P21 and P27 in squamous carcinoma cell lines of the head and neck.
1,25(OH)2 vitamin D3 induces elevated expression of the cell cycle-regulating genes P21 and P27 in squamous carcinoma cell lines of the head and neck.
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DOI：
10.1080/000164801300006353
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发表时间：
2001
影响因子：
1.4
通讯作者：
G. Hager;M. Formanek;C. Gedlicka;D. Thurnher;B. Knerer;J. Kornfehl
中科院分区：
文献类型：
--
作者：
G. Hager;M. Formanek;C. Gedlicka;D. Thurnher;B. Knerer;J. Kornfehl
维生素D3的生物活性形式1，25-二羟基维生素D3 [1，25（OH）2D 3]抑制各种恶性细胞的增殖并诱导分化，包括头颈部鳞状细胞癌细胞系（SCCHN）。这些作用是由于细胞停滞在细胞周期的G 0/G1期，主要由维生素D受体介导。为了进一步探讨SCCHN中抗增殖活性的分子机制，我们研究了1，25（OH）2D 3对G1期调节蛋白cyclin D1、p21和p27表达的影响。此外，作为G1蛋白复合物的直接靶标，我们研究了视网膜母细胞瘤蛋白（pRb）的磷酸化状态。将2种SCCHN细胞系[JPPA（喉癌）和SCC 9（舌癌）]和人永生化角质形成细胞（HaCaT）的同步化细胞在存在或不存在（乙醇作为对照）1，25（OH）2D 3（10（-7）M）的情况下培养96小时。在不同的时间间隔，细胞周期状态检测荧光激活细胞分选（FACS）分析和平行的细胞周期调节蛋白的表达，确定在蛋白质和mRNA水平。在所有测试的细胞系中，1，25（OH）2D 3导致细胞停滞在细胞周期的G 0/G1期，并显著诱导抑制剂p21和p27的表达。对cyclin D1的表达无明显影响。p21和p27 mRNA的诱导揭示了维生素D受体的转录调控。同时，过度磷酸化的pRb被转化为低磷酸化的形式。我们的研究结果表明，维生素D3的生物活性形式直接调节p21和p27的表达，诱导G 0/G1期阻滞：1，25（OH）2D 3控制SCCHN细胞增殖的一种机制。
The biologically active form of vitamin D3, 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], inhibits proliferation and induces differentiation for various malignant cells, including squamous cell carcinoma cell lines of the head and neck (SCCHN). These effects are due to an arrest of cells in the G0/G1 phase of the cell cycle and are predominantly mediated by the vitamin D receptor. To further explore the molecular mechanisms of the antiproliferative activity in SCCHN we studied the influence of 1,25(OH)2D3 on the expression of the G1 phase-regulating proteins cyclin D1, p21 and p27. Furthermore, as a direct target of G1 protein complexes, we investigated the phosphorylation status of the retinoblastoma protein (pRb). Synchronized cells of 2 SCCHN cell lines [JPPA (laryngeal carcinoma) and SCC 9 (tongue carcinoma)] and human immortalized keratinocytes (HaCaT) were cultured for 96 h in the presence or absence (ethanol as control) of 1,25(OH)2D3 (10(-7) M). At various time intervals the cell cycle status was detected by fluorescence-activated cell sorting (FACS) analysis and in parallel the expression of cell cycle-regulating proteins was determined at the protein and mRNA levels. In all cell lines tested 1,25(OH)2D3 caused an arrest of cells in the G0/G1 phase of the cell cycle and markedly induced the expression of the inhibitors p21 and p27. No influence was detectable on the expression of cyclin D1. Induction of p21 and p27 mRNA revealed transcriptional regulation by the vitamin D receptor. Simultaneously, hyperphosphorylated pRb was transformed to the hypophosphorylated form. Our results demonstrate that the biologically active form of vitamin D3 directly regulates the expression of p21 and p27, inducing a G0/G1 phase arrest: one mechanism by which 1,25(OH)2D3 controls cell proliferation inSCCHN.