Differentiation of human fetal osteoblastic cells and gap junctional intercellular communication
Differentiation of human fetal osteoblastic cells and gap junctional intercellular communication
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DOI：
10.1152/ajpcell.2000.278.2.c315
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发表时间：
2000-02-01
影响因子：
5.5
通讯作者：
Yellowley, CE
中科院分区：
文献类型：
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作者：
Donahue, HJ;Li, ZY;Yellowley, CE
间隙连接通道促进细胞间通讯，并在这样做可能有助于细胞分化。为了验证这一假设，我们研究了温度敏感的人胎儿成骨细胞系（hFOB 1.19），当在37摄氏度培养时迅速增殖，但当在39.5摄氏度培养时增殖缓慢，并显示增加碱性磷酸酶活性和骨钙素合成的间隙连接的表达和功能。我们发现hFOB 1.19细胞表达丰富的连接蛋白43（Cx43）蛋白和mRNA。而Cx45 mRNA表达较低，Cx 26和Cx 32 mRNA未检测到。相对于37 ℃，在39.5 ℃培养hFOB 1.19细胞抑制增殖，增加Cx43 mRNA和蛋白表达，并增加间隙连接细胞间通讯（GJIC）。用18 α-大黄酸阻断GJIC可防止39.5 ℃培养引起的碱性磷酸酶活性增加，但不影响骨钙素水平。这些结果表明，间隙连接的功能和表达平行。成骨细胞的分化，并有助于碱性磷酸酶活性的表达，碱性磷酸酶活性是完全分化的成骨细胞的标志物。
Gap junctional channels facilitate intercellular communication and in doing so may contribute to cellular differentiation. To test this hypothesis, we examined gap junction expression and function in a temperature-sensitive human fetal osteoblastic cell line (hFOB 1.19) that when cultured at 37 degrees C proliferates rapidly but when cultured at 39.5 degrees C proliferates slowly and displays increased alkaline phosphatase activity and osteocalcin synthesis. We found that hFOB 1.19 cells express abundant connexin 43 (Cx43) protein and mRNA. In contrast, Cx45 mRNA was expressed to a lesser degree, and Cx26 and Cx32 mRNA were not detected. Culturing hFOB 1.19 cells at 39.5 degrees C, relative to 37 degrees C, inhibited proliferation, increased Cx43 mRNA and protein expression, and increased gap junctional intercellular communication (GJIC). Blocking GJIC with 18 alpha-glycyrrhetinic acid prevented the increase in alkaline phosphatase activity resulting from culture at 39.5 degrees C but did not affect osteocalcin levels. These results suggest that gap junction function and expression parallel. osteoblastic differentiation and contribute to the expression of alkaline phosphatase activity, a marker for fully differentiated osteoblastic cells.