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Blocking gastrin and CCK-B autocrine loop affects cell proliferation and apoptosis in vitro

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Abstract

Gastrin and cholecystokinin-B receptor (CCK-B) were co-expressed in human gastric carcinoma tissues, suggesting that a functional autocrine loop, the gastrin and CCK-B receptor loop, may be presented in gastric cancer cells and play an important role in the pathogenesis and progression of gastric carcinomas. The present study was aimed at studying the effects of blocking the gastrin and CCK-B receptor loop on cell proliferation and apoptosis in gastric cancer cell line SGC-7901 cells (SGC-7901 cells). First, the expression of gastrin and CCK-B receptor mRNAs and gastrin protein in SGC-7901 cells were measured by RT-PCR and immunocytochemistry, respectively. Radioimmunoassay (RIA) was used to detect the concentrations of gastrin in culture medium. The gastrin–CCK-B receptor axis was blocked by using a specific neutralizing antibody against human gastrin and siRNA specifically targeting human CCK-B receptors, respectively. Flow cytometry was used to measure the cell cycle and apoptotic cells, and western blotting was used to measure the expression of CCK-B receptor, caspase-3, and matrix metalloproteinase-2 (MMP-2) in cells. The results showed that SGC-7901 cells not only coexpressed gastrin and CCK-B receptor mRNAs, but also endogenously secreted gastrin protein into the culture medium, thus forming gastrin–CCK-B receptor autocrine loop. Biologically, disrupting gastrin–CCK-B receptor autocrine loop by neutralizing the endogenous gastrin or by knocking down CCK-B receptor expression significantly inhibited the cell proliferation and decreased the percentage of cells residing in the S-phase of the cell cycle, and meanwhile promoted cell apoptosis and increased caspase-3 expression as well as decreased MMP-2 expression. An autocrine loop between endogenously secreted gastrin and CCK-B receptors may play a key role in the regulation of cell proliferation and apoptosis in SGC-7901 cells.

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Acknowledgment

We are grateful to Dr. Guo YangBiao (Department of Biochemistry and Molecular Biology, School of Basic Medical Science, Sichuan University, Chengdu, China) and Yan WenXiu (Department of Pathology, Guiyang Medical University) for their many contributions to this paper. This work was supported by National Natural Science Foundation of China, No. 30560038, the Science and Technology Foundation of GuiZhou Province (No. [2007] 2084), and the Project of International Cooperation in Science and Technology of Guizhou Province (No. [G] 700119).

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Authors and Affiliations

  1. Affiliated Hospital and Molecular Biology Key Laboratory, Guiyang Medical University, No. 9 Beijiang Road, Guiyang, 550004, GuiZhou, China

    Jian Jiang Zhou

  2. Department of Biochemistry and Molecular Biology of School of Basic Medical Science, Sichuan University, Chengdu, 610041, Sichuan Province, China

    Man Ling Chen

  3. Center for Craniofacial Molecular Biology, School of Dentistry, University of Southern California, Los Angeles, CA, USA

    Qun Zhou Zhang

  4. Molecular Biology Key Laboratory of Guiyang Medical University, Guiyang, 550004, Guizhou Province, China

    Yan Zao & Yuan Xie

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  1. Jian Jiang Zhou
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Correspondence to Jian Jiang Zhou.

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Zhou, J.J., Chen, M.L., Zhang, Q.Z. et al. Blocking gastrin and CCK-B autocrine loop affects cell proliferation and apoptosis in vitro. Mol Cell Biochem 343, 133–141 (2010). https://doi.org/10.1007/s11010-010-0507-5

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  • DOI: https://doi.org/10.1007/s11010-010-0507-5

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