Proteomics provides insights into the inhibition of Chinese hamster V79 cell proliferation in the deep underground environment

Abstract

As resources in the shallow depths of the earth exhausted, people will spend extended periods of time in the deep underground space. However, little is known about the deep underground environment affecting the health of organisms. Hence, we established both deep underground laboratory (DUGL) and above ground laboratory (AGL) to investigate the effect of environmental factors on organisms. Six environmental parameters were monitored in the DUGL and AGL. Growth curves were recorded and tandem mass tag (TMT) proteomics analysis were performed to explore the proliferative ability and differentially abundant proteins (DAPs) in V79 cells (a cell line widely used in biological study in DUGLs) cultured in the DUGL and AGL. Parallel Reaction Monitoring was conducted to verify the TMT results. γ ray dose rate showed the most detectable difference between the two laboratories, whereby γ ray dose rate was significantly lower in the DUGL compared to the AGL. V79 cell proliferation was slower in the DUGL. Quantitative proteomics detected 980 DAPs (absolute fold change ≥ 1.2, p < 0.05) between V79 cells cultured in the DUGL and AGL. Of these, 576 proteins were up-regulated and 404 proteins were down-regulated in V79 cells cultured in the DUGL. KEGG pathway analysis revealed that seven pathways (e.g. ribosome, RNA transport and oxidative phosphorylation) were significantly enriched. These data suggest that proliferation of V79 cells was inhibited in the DUGL, likely because cells were exposed to reduced background radiation. The apparent changes in the proteome profile may have induced cellular changes that delayed proliferation but enhanced survival, rendering V79 cells adaptable to the changing environment.

First transcriptome profiling of D. melanogaster after development in a deep underground low radiation background laboratory

“…The input of low background was estimated for the first time for this multicellular complex organism by comparing the results of RNA-seq for flies developed in natural radiation background and in low background conditions. It is important to note that at the moment there is no consensus about the effect of low-radiation background on living organisms—in some experiments the absence of its influence was recorded [ 13 , 16 ], in others the effect of background radiation reduction was observed [ 6 , 12 , 14 , 24 ]. But biological parameters evaluated in different experiments for various model organisms were so diverse and multidirectional that they do not allow making any general conclusion [ 11 , 20 , 52 ].…”

“…Effects of background ionizing radiation deprivation on organisms have been studied for the last two decades [ 6 , 11 – 17 ] in experiments performed in various locations, for instance, in the low-background chamber at Osaka University (Osaka, Japan) and several industrial underground tunnels, such as DUGL CJEM (Erdaogou Mine, China), BISAL (Boulby, UK) and WIPP (New Mexico, USA) [ 12 , 14 , 16 , 18 ]. But most of biological studies dealt with deep underground laboratories, namely: LNGS (Gran Sasso, Italy), LSM CNRS (Modane, France), SNOLAB (Sudbury, Canada), JINR/BNO INR RAS (Neutrino village, Russia) [ 7 , 11 , 15 , 17 , 19 – 21 ].…”

“…The current results were obtained in such experiments when model organisms were exposed to low-background conditions from several days to about one year [ 11 , 14 , 20 , 22 ]. Effects of low-radiation background on living organisms were determined by comparing experimental and control groups and by registering the changes in parameters suitable for experimental purposes, for instance, growth rate, lifespan, fertility, gene expression or protein abundance [ 12 – 15 , 20 ]. An additional difficulty in carrying out such experiments is the demand to control many external parameters which must be the same for groups of organisms placed in low-background conditions and for those left in natural background conditions.…”

“…At present the significant part of the effects registered in decreased background radiation conditions can be classified as suppressive, that reduce growth and development parameters, both for unicellular and multicellular organisms. Growth rate inhibition, increased sensitivity to radiation, delayed recovery after return to natural background conditions, alteration in expression of several genes concerned with ribosomal proteins, membrane transport, respiration, reparation and antioxidant regulation for increased ROS removal were observed in experiments on mammalian cell cultures (TK6, V79, M10, L5178Y, FD-LSC-1) and unicellular organisms ( Paramecium tetraurelia , Synechococcus lividus , Shewanella oneidensis , Deinococcus radiodiodurans , Saccharomyces cerevisiae ) [ 11 , 12 , 14 , 17 , 18 , 24 , 25 ]. The first multicellular organism used for determination of the effects of low-radiation background was the fruit fly in projects FLYINGLOW and RENOIR [ 20 , 26 ].…”

“…It is important to note that many researchers who have conducted biological experiments in low-background laboratories, for instance, SNOLAB and DUGL CJEM, point out that some factors, not related to radiation, influence biological responses, which further complicates the analysis of the obtained data [ 12 , 15 ].…”