Li Fei, Zhang Zong-Ming
Department of General Surgery, Digestive Medical Center, The First Affiliated Hospital, Medical School, Tsinghua University, Beijing, China.
World J Gastroenterol. 2009 Jun 28;15(24):3046-50. doi: 10.3748/wjg.15.3046.
To identify and compare the profile of Ca(2+) channel subunit expression in INS-1 and rat pancreatic beta cells.
The rat insulin-secreting INS-1 cell line was cultured in RPMI-1640 with Wistar rats employed as islet donors. Ca(2+) channel subunit expression in INS-1 and isolated rat beta cells were examined by reverse transcription polymerase chain reaction (RT-PCR). Absolute real-time quantitative PCR was performed in a Bio-Rad iQ5 Gradient Real Time PCR system and the data analyzed using an iQ5 system to identify the expression level of the Ca(2+) channel subunits.
In INS-1 cells, the L-type Ca(2+) channel 1C subunit had the highest expression level and the TPRM2 subunit had the second highest expression. In rat beta cells, the TPRC4beta subunit expression was dominant and the expression of the L-type 1C subunit exceeded the 1D subunit expression about two-fold. This result agreed with other studies, confirming the important role of the L-type 1C subunit in insulin-secreting cells, and suggested that non-voltage-operated Ca(2+) channels may have an important role in biphasic insulin secretion.
Twelve major Ca(2+) channel subunit types were identified in INS-1 and rat beta cells and significant differences were observed in the expression of certain subunits between these cells.
鉴定并比较INS-1细胞和大鼠胰腺β细胞中Ca(2+)通道亚基的表达情况。
将大鼠胰岛素分泌细胞系INS-1培养于RPMI-1640培养基中，以Wistar大鼠作为胰岛供体。采用逆转录聚合酶链反应（RT-PCR）检测INS-1细胞和分离的大鼠β细胞中Ca(2+)通道亚基的表达。在Bio-Rad iQ5梯度实时荧光定量PCR系统中进行绝对实时定量PCR，并使用iQ5系统分析数据，以确定Ca(2+)通道亚基的表达水平。
在INS-1细胞中，L型Ca(2+)通道1C亚基表达水平最高，TPRM2亚基表达水平次之。在大鼠β细胞中，TPRC4β亚基表达占主导，L型1C亚基的表达量约为1D亚基表达量的两倍。该结果与其他研究一致，证实了L型1C亚基在胰岛素分泌细胞中的重要作用，并提示非电压门控Ca(2+)通道可能在胰岛素双相分泌中起重要作用。
在INS-1细胞和大鼠β细胞中鉴定出12种主要的Ca(2+)通道亚基类型，且这些细胞中某些亚基的表达存在显著差异。
