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Real-time cell analysis and heat shock protein gene expression in the TcA Tribofium castaneum cell line in response to environmental stress conditions

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Insect Science 2017年第3期24卷 358-370页

作者： Andres Garcia-Reina Maria Juliana Rodriguez-Garcia guillermo ramis Jose Galian Department of Zoology and Physical Anthropology University of MurciaCampus Mare Nostrum E-30100 Murcia Spain Department of Animal Production Faculty of Veterinary University of MurciaCampus Mare Nostrum E-30100 Murcia Spain

The rust red flour beetle, Tribolium castaneum （Herbst, 1797）（Coleoptera： Tenebrionidae）, is a pest of stored grain and one of the most studied insect model species. Some of the previous studies involved heat response studies in terms of survival and heat shock protein expression, which are regulated to protect other proteins against environ- mental stress conditions. In the present study, we characterize the impedance profile with the xCELLigence Real-Time Cell Analyzer and study the effect of increased temperature in cell growth and viability in the cell line BCIRL-TcA-CLG 1 （TcA） of T castaneum. This novel system measures cells behavior in real time and is applied for the first time to insect cells. Additionally, cells are exposed to heat shock, increased salinity, acidic pH and UV-A light with the aim of measuring the expression levels of lisp27, Hsp68a, and Hsp83 genes. Results show a high thermotolerance of TeA in terms of cell growth and viability. This result is likely related to gene expression results in which a significant up-regulation of all studied Hsp genes is observed after 1 h of exposure to 40 ~C and UV light. All 3 genes show similar expression patterns, but Hsp27 seems to be the most affected. The results of this study validate the RTCA method and reveal the utility of insect cell lines, real-time analysis and gene expression studies to better understand the physiological response of insect cells, with potential applications in different fields of biology such as conservation biology and pest management.

关键词： environmental stress gene expression heat shock proteins insect cells,real-time cell analysis (RTCA) red flour beetle

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